Dissertations / Theses on the topic 'Protein kinase CK2 – Pathophysiology'
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Holland, Zoe. "Plasmodium falciparum protein kinase CK2." Thesis, University of Glasgow, 2008. http://theses.gla.ac.uk/606/.
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Malaria, caused by infection with intracellular protozoan parasites of the genus Plasmodium, is responsible for 300 to 600 million clinical cases annually (Snow et al., 2005), resulting in the deaths of up to three million people every year (Breman, 2001, Breman et al., 2004). There is a clear need for further research aimed at identifying novel drug targets (Ridley, 2002). Reversible phosphorylation of proteins is a major regulatory mechanism in most cellular processes, and protein kinases are considered promising drug targets, comprising as much as 30% of all protein targets under investigation (Cohen, 2002). The divergences between human and plasmodial protein kinases suggest that specific inhibition of the latter is an achievable goal (Doerig, 2004, Doerig and Meijer, 2007). This study investigates protein kinase CK2 of Plasmodium falciparum, seeking to establish by reverse genetics and biochemical approaches whether it represents a possible antimalarial drug target. Protein-kinase CK2, formerly known as Casein Kinase II, is a dual-specificity (Serine/Threonine and Tyrosine) protein kinase ubiquitously expressed in eukaryotes. It has over 300 cellular substrates catalogued to date (Meggio and Pinna, 2003). Consistent with its multiple substrates, the enzyme plays a crucial role in many cellular processes, and is essential to viability in yeast and slime mould (Padmanabha et al., 1990, Kikkawa et al., 1992). The human CK2 holoenzyme consists of two catalytic a or a’ subunits and two regulatory b subunits, and recent evidence indicates that the latter interact with several protein kinases in addition to CK2a (reviewed in (Bibby and Litchfield, 2005)), pointing to a likely role in the integration of numerous signalling pathways. A putative CK2a orthologue and two predicted CK2b subunits were identified in the P. falciparum genome (Ward et al., 2004, Anamika et al., 2005). Here we present the biochemical characterisation of the PfCK2a orthologue and both PfCK2b orthologues, and demonstrate by using a reverse genetics approach that each of the three subunits is essential for completion of the erythrocytic asexual cycle of the parasite, thereby validating the enzyme as a possible drug target. Recombinant PfCK2a possesses protein kinase activity, exhibits similar substrate and co-substrate preferences to those of CK2a subunits from other organisms, and interacts with both of the PfCK2b subunits in vitro. PfCK2a is amenable to inhibitor screening, and we report differential susceptibility between the human and P. falciparum CK2a enzymes to a small molecule inhibitor. Taken together, the data indicate that PfCK2a is an attractive, validated target for antimalarial chemotherapeutic intervention.
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Ling, Ann Lee. "Protein kinase CK2 : structure, interactions and inhibition." Thesis, University of Cambridge, 2011. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.609645.
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Ganley, Ian Gordon. "Interaction of phospholipase D1 with protein kinase CK2." Thesis, University of Cambridge, 2002. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.620410.
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Heriche, Jean-Karim. "Protéine kinase CK2 et prolifération cellulaire." Grenoble 1, 1996. http://www.theses.fr/1996GRE10174.
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La proteine kinase ck2 est une serine/threonine kinase dont la sous-unite catalytique (ck2a) est souvent associee a une sous-unite regulatrice (ck2b) au sein d'un tetramere a#2b#2. Bien que sa fonction et sa regulation ne soient pas connues, on suppose qu'elle participe au controle de la proliferation cellulaire. Un signal mitogene majeur dans de nombreux types cellulaires est forme par l'activation sequentielle des trois kinases raf/mek1/mapk. Nous nous sommes donc demandes si la ck2 pouvait etre connectee a cette voie de transduction du signal proliferatif. Pour cela, nous avons suivi une demarche a base de transfections et d'experiences in vitro en utilisant des mutants des proteines impliquees. Nous avons ainsi pu montrer qu'une population de ck2a independante de ck2b presente les caracteristiques d'un nouveau substrat pour raf. En effet, ck2a est phosphorylee et inhibee en presence de raf actif in vitro et in vivo et la presence de ck2b empeche cette phosphorylation. Nous avons egalement montre que ck2a peut s'associer a mek1 et inhiber son activation par le serum. Cet effet de ck2a sur mek1 passe par l'association de ck2a avec la phosphatase pp2a car un mutant de ck2a incapable de s'associer a la phosphatase n'a plus d'effet sur mek1. L'inhibition de mek1 par ck2a correle avec une forte inhibition de la croissance clonale des cellules en presence de ck2a. De plus, le complexe ck2a/pp2a est dissocie en reponse au pdgf in vivo et par raf in vitro, suggerant que ce complexe est bien un regulateur physiologique de mek1 et de la proliferation cellulaire. Enfin, en exploitant les analogies entre ck2a et la serine/threonine kinase apparentee gsk-3, nous avons egalement trouve que ck2a pouvait etre un substrat associe de la tyrosine kinase abl. La signification biologique de cette association est discutee
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Wang, Zilong. "Expression of epitope-tagged protein kinase CK2 in mammalian cells." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1998. http://www.collectionscanada.ca/obj/s4/f2/dsk2/ftp01/MQ32281.pdf.
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Lee, Yew. "Molecular and transgenic approaches to understanding the function of protein kinase CK2 in plants /." Digital version accessible at:, 1998. http://wwwlib.umi.com/cr/utexas/main.
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Hou, Zengye. "Development of Novel Protein Kinase CK2 Inhibitors with Nitrogen Heterocyclic Scaffolds." 京都大学 (Kyoto University), 2013. http://hdl.handle.net/2433/174543.
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Mitchell, Louise E. "The regulation of RNA polymerase III transcription by protein kinase CK2." Thesis, University of Glasgow, 2008. http://theses.gla.ac.uk/399/.
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In order for cells to proliferate, a certain size has to be reached, which depends primarily on the rate of translation. RNA polymerase (pol) III plays a key role in protein synthesis by catalysing the production of small, untranslated RNA molecules such as transfer (tRNA) and 5S ribosomal RNA (5S rRNA). Indeed, recent evidence suggests that tRNAiMet production is limiting for translation and proliferation in some cell types. Therefore, the rate of pol III transcription plays a fundamental role in cellular growth and proliferation. Regulation of pol III output is mediated via a number of different mechanisms that can alter the activities of the transcription factors which are responsible for directing pol III transcription. Work presented in this thesis aimed at investigating the mechanisms behind the regulation of pol III transcription by the protein kinase CK2.
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Llinas, Alexander J. "The activities of protein kinase CK2 in oogenesis of Xenopus laevis." Thesis, University of St Andrews, 1999. http://hdl.handle.net/10023/14510.
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Protein kinases play important roles in regulating cellular functions in many organisms. This work deals specifically with the protein kinase CK2 (casein kinase II) and its role in regulating the activity of proteins involved in oocyte development in Xenopus laevis. Protein kinase CK2 is a tetrameric enzyme containing two catalytic subunits (alpha and alpha') and two identical regulatory subunits (beta) which forms the holoenzyme. CK2 phosphorylates many different proteins involved in many aspects of cellular functions. It phosphorylates serine and threonine sites and is considered to be a ubiquitous enzyme, expressed at different levels in different cell types. In this study CK2 activity was characterized in material from two sources: from isolated nuclei and messenger ribonucleoprotein (mRNP) particles from Xenopus oocytes. cDNAs expressing both the alpha and the beta subunits were cloned and antibodies were raised against the fusion protein containing the beta-subunit. The main objective of this study was to determine the effects that CK2 had on proteins involved in oocyte development. The interaction of CK2 with a protein known as histone deacetylase was studied in depth to determine how phosphorylation might influence its function and cellular compartmentalisation. Specifically, phosphorylation by CK2 is shown to improve the kinetics of nuclear unport, and the interaction of histone deacetylase with alpha-importin, a well-established nuclear transport protein, is revealed to be dependent on the phosphorylation state of histone deacetylase. Another aspect of this work is related to the association of CK2 with mRNP particles in the cytoplasm. mRNP particles function as long term storage units for mRNA to be used during oocyte maturation and early embryogenesis. It has been postulated that a protein kinase associated with these particles plays a role in controlling the binding of mRNA to proteins involved in translation repression (mRNA masking proteins). This study lends support to that theory, and the possible effects of CK2 phosphorylation on the masking "Y-box" proteins are discussed.
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Bestgen, Benoit. "Selective modulation of the Protein Kinase CK2 : discovery, syntheses and characterization of non-ATP site inhibitors of CK2." Thesis, Lyon 1, 2015. http://www.theses.fr/2015LYO10247.
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La Protéine Kinase CK2 est une enzyme tétramérique composé d'un dimère de sous-unité régulatrice (β) et de deux sous-unités catalytiques, CK2α et/ou CK2α'. La sous-unité catalytique de CK2 est constitutivement active alors que la sous-unité régulatrice régule seulement la sélection des substrats phosphorylés par CK2. CK2 est une Ser/Thr protéine kinase ubiquitaire impliquée dans le contrôle de nombreuses voies de signalisations. La dérégulation de CK2 promeut le développement des cancers et il a été démontré que CK2 est une cible pertinente dans le traitement des cancers. Notre objectif était de cibler la protéine kinase CK2 de manière indépendante du site actif. Deux séries de composés ont été étudiés : Basé sur un premier hit faiblement actif (CI50 = 30 μM) mais inhibant CK2 de manière non- ATP compétitive, des dérivés comportant le noyau 2-aminothiazole ont été synthétisés et un composé actif (CI50 = 0,6 μM) et efficace in cellulo a été obtenu. Grace à des expériences sur des mutants ponctuels de CK2, des expériences de dichroïsme circulaire, de la STD-RMN et de la modélisation moléculaire, le site de fixation de nos inhibiteurs a été précisément défini à l'interface de la boucle riche en glycine et de l'hélice-αC. Des inhibiteurs de l'interaction α/β ont été étudiés à partir d'un peptide cyclique jusqu'au développement de petites molécules via un screening virtuel. Des études de relations structure-activité ont été réalisé sur la série de composés synthétisées et des tests cellulaires ont été mis en place afin d'évaluer ces composés. Les deux classes de molécules décrites sont des outils intéressants pour comprendre la régulation physiologique de la protéine kinase CK2 et des opportunités prometteuses dans le traitement de certains cancersThe protein kinase CK2 is a tetrameric enzyme composed of a dimer of regulatory subunits (β) and two catalytic subunits, CK2α and/or CK2α’. The catalytic subunit of CK2 is constitutively active, while the regulatory subunit modulates the selectivity toward a subset of substrate proteins. CK2 is a ubiquitous Ser/Thr protein kinase involved in the control of various signaling pathways, and dysregulation of CK2 promotes cancer development. CK2 has been proved to be a valuable target in cancer treatment. Our objective was to target CK2 outside the ATP-pocket. Two independent classes of compounds were studied: Based on a first hit with a low potency (IC50 = 30 μM) but a non-ATP competitive mechanism of action, several 2-aminothiazole derivatives were synthesized to lead to a potent (IC50 = 0.6 μM) and cell efficient allosteric inhibitor of CK2. Using single mutation scanning, CD spectrometry, STD-NMR and docking experiments, the binding site of our compounds was precisely defined outside the ATP-pocket, at the interface of the glycine-rich loop and the αC-helix. Inhibitors of the α/β interaction were studied from a small cyclic peptide to the development of small molecules through Virtual Ligand Screening. Structures Activity Studies were conducted on the synthesized derivatives and cellular based assays to evaluate the α/β inhibitors were set up. The two classes of compounds developed herein are valuable tools to understand the physiological regulation of the protein kinase CK2, and potential new opportunities in cancer treatment
Das Protein Kinase CK2 ist ein tetrameres Enzym, das aus einem Dimer von regulatorischen Untereinheiten (β) und zwei katalytischen Untereinheiten (CK2α und/oder CK2α’) besteht. Die katalytische Untereinheit der CK2 ist konstitutiv aktiv, während die regulatorische Untereinheit die Auswahl einiger der durch CK2 phosphorylierten Substrate steuert. CK2 ist eine ubiquitäre Proteinkinase, die an der Kontrolle zahlreicher Signalwege beteiligt ist. Eine Fehlregulation der CK2 fördert die Tumorenstehung. Es konnte gezeigt werden, dass CK2 eine vielversprechende Zielstruktur für die Entwicklung neuer Therapeutika ist. Unser Ziel war es, neue Hemmstoffe der Proteinkinase CK2 zu entwickeln, die an anderen Stellen als dem aktiven Zentrum angreifen. Zwei Serien von Verbindungen sind untersucht worden: Basierend auf einem ersten schwach aktiven “Hit” (IC50 = 30 μM), der einen nicht-ATPkompetitiven Wirkmechanismus aufwies, wurden einige neue 2-Aminothiazol-Derivate synthetisiert. Dadurch wurden allosterische Inhibitoren mit einer deutlich gesteigerte Potenz (IC50 = 0,6 μM) und einer beachtlichen Zellaktivität erhalten. Mittels eines CK2- Punktmutanten-Screenings, Zirkulardichroismus-Spektrometrie, STD-NMR und molekularer Docking-Simulationen konnte die Bindestelle unserer Hemmstoffe außerhalb der ATPBindetasche, zwischen der Glycin-reichen Schleife und der αC-Helix, lokalisiert werden. Desweiteren wurden niedermolekulare Inhibitoren der α/β-Interaktion entwickelt, ausgehend von einem zyklischen Peptid sowie von Hitverbindungen aus einem virtuellen Screening. Neue Verbindungen wurden synthetisiert und die Struktur- Wirkungsbeziehungen analysiert; zusätzlich wurde ein Zellassay zur Überprüfung des postulierten Wirkmechanismus etabliert. Die beiden entwickelten Verbindungsklassen sind interessante Werkzeuge, um die physiologische Regulation der Proteinkinase CK2 näher zu analysieren; überdies stellen sie Ausgangspunkte für die Entwicklung neuartiger Krebstherapeutika dar
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Armengot, Martínez Laia. "Unraveling new roles and substrates for protein kinase CK2 in arabidopsis thaliana." Doctoral thesis, Universitat Autònoma de Barcelona, 2014. http://hdl.handle.net/10803/285612.
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Aquesta tesi és part d’un projecte d’investigació més general que té com a objectiu estudiar el paper de la serina/treonina quinasa CK2 en el desenvolupament de les plantes, utilitzant Arabidopsis thaliana com a model. Malgrat ser una de les primeres quinases identificades, les vies de senyalització en què participa la CK2 encara no estan completament caracteritzades.
En la primera part d’aquesta tesi es descriu la implicació de la quinasa CK2 en la via de senyalització de l’àcid salicílic (SA) i, el control exercit per aquesta hormona en l’expressió dels gens que codifiquen per les proteïnes PIN (exportadores d’auxina), i per a la quinasa reguladora d’aquests transportadors, la proteïna PINOID (PID). Antics membres del laboratori on s’ha realitzat aquesta tesi havien previament obtingut un mutant dominant negatiu de la CK2 (CK2mut) en Arabidopsis (Moreno-Romero et al., 2008). En aquest treball mostrem que aquestes plantes contenen nivells elevats de SA i que l’acumulació de SA a les arrels és el responsable del fenotip radicular alterat (disminucio de la longitud de l’arrel i absència d’arrels lateral). També demostrem que tractaments amb SA exogen redueixen la transcripció dels gens que codifiquen per als transportadors PIN1-PIN4 i PIN7, mentre que estimulen la transcripció del gen que codifica per la quinasa PID. Aquest efecte és similar a l’observat en les arrels de les plantes CK2mut, exceptuant als gens PIN4 i PIN7 que es mostren sobreexpressats. Aquest resultat, suggereix que l’efecte repressor de SA en l’expressió de PIN4 i PIN7 requereix que la quinasa CK2 sigui funcional. D’altra banda, SA estimula l’expressió de les subunitats de CK2, mentre que la pèrdua d’activitat CK2 a les plantes CK2mut produeix un augment de la transcripció de gens relacionats amb la biosíntesi de SA. Aquests resultats suggereixen l'existència d’un loop de retroalimentació negatiu entre la CK2 i el SA, necessari per mantenir la homeostasi de SA. En aquest capítol també es mostra que la sobreexpressió d’una subunitat CK2α catalíticament activa provoca un increment del sistema radicular de les plantes.
En la segona part d’aquesta tesi, ens hem centrat en la recerca i caracterització de substrats de la CK2 en Arabidopsis. Amb aquest objectiu, vam realitzar un assaig de doble híbrid en llevat, que ens va permetre identificar, entre d’altres, a la proteïna NPH3 com a possible substrat de la CK2. NPH3 és un element essencial de la via de senyalització del fototropisme; la seva activitat en aquesta via depèn del seu estat de fosforilació i del seu paper com a adaptador de substrat d’un complex d’ubiquitinació del tipus Cullin3-Ring E3 lligasa (CRL3NPH3). Aquest complex ubiquitina al fotoreceptor de llum blava phototropin 1, el qual es troba associat a la membrana plasmàtica. En la foscor, la proteïna NPH3 està fosforilada i inactiva, mentre que en condicions de llum, està defosforilada i activa, i dirigeix la ubiquitinació de phot1. Recentment, s’ha proposat que la ubiquitinació de phot1 promou la seva internalització des de la membrana cap al citoplasma. Els resultats obtinguts en aquest treball demostren que la quinasa CK2 fosforila NPH3 in vitro i que l’activitat d’aquesta quinasa és necessària per a la fosforilació in vivo en condicions de foscor. A més, la fosforilació de NPH3 per part de la CK2 és important per a mantenir la proteïna NPH3 inactiva. D’altra banda, la manca d’activitat CK2 provoca l’internalització de phot1, inclús en foscor, fet que podria estar relacionat amb el fenotip no fototròpic de les plantes sense activitat CK2. Aquesta internalització és independent de la presència de NPH3 i, per tant, independent d’ubiquitinació. Sorprenentment, la internalització de phot1 observada en condicions de llum, també és independent de la presència de NPH3.This thesis is part of a research project that aims to study the role of the serine/threonine protein kinase CK2 in plant development, using Arabidopsis thaliana as a model. Despite being one of the first kinases identified, the signaling pathways in which CK2 is involved are not yet fully characterized. The first part of this thesis describes the involvement of CK2 in the signaling pathway of salicylic acid (SA), and the control exercised by this hormone in the expression of genes coding for auxin membrane transporters (the PIN proteins) and their regulatory kinase PINOID (PID). Former members of the group where this thesis was carried out had obtained a dominant negative mutant of CK2 (CK2mut plants). These plants showed altered root phenotypes (decrease of the main root length and absence of lateral root formation) and changes in the transcription levels of genes encoding several of the PIN proteins (PIN1-PIN4 and PIN7) and of the kinase PINOID (PID) (Marques-Bueno et al., 2011). Here, we show that CK2mut plants contain high levels of salicylic acid, which are responsible for the root phenotype of CK2mut plants. We also demonstrate that treatment of Arabidopsis wild-type plants with exogenous SA inhibits the transcription of genes coding for proteins PIN1-PIN4 and PIN7, while it stimulates the transcription of the PID encoding gene. This effect is similar to that observed in roots of CK2mut plants, except for PIN4 and PIN7 genes, which are overexpressed, suggesting that the repressive effect of SA on PIN4 and PIN7 expression requires a functional CK2. Moreover, SA stimulates the expression of CK2 subunits, whereas the loss of CK2 activity in CK2mut plants produces an increase in the transcript levels of genes related to SA biosynthesis. We propose the existence of a negative feedback loop between CK2 and SA, needed to maintain the homeostasis of SA. This chapter also shows that overexpression of a catalytically active α subunit of CK2 improves the root system of Arabidopsis plants. The second part of this thesis focuses on the searching and characterization of plant CK2 substrates. For this purpose, we performed a large scale yeast two-hybrid screen that resulted in the identification of 28 potential CK2 substrates. Among them, we found four members of the same protein family, called NPH3/RPT2 (NRL), including NPH3, the founder member of the family. NPH3 is an essential element of the phototropic signaling pathway, and its activity in this pathway depends on its phosphorylation state and on its role as a substrate adapter within the Cullin3-Ring E3 ligase (CRL3NPH3) ubiquitination complex. CRL3NPH3 ubiquitinates the membrane-associated blue light photoreceptor phototropin 1. In the dark, NPH3 is phosphorylated and inactive, while in light conditions it is defosforilated and active and directs ubiquitination of phot1. Recently, it has been proposed that ubiquitination of phot1 promotes its internalization from the plasma membrane into the cytoplasm. Here we show that CK2 phosphorylates NPH3 in vitro, and that CK2 activity is required for the in vivo NPH3 phosphorylation in darkness. In addition, phosphorylation of NPH3 by CK2 is important to keep the protein inactive. Moreover, we observe that the lack of CK2 activity causes internalization of phot1 even in darkness, which could be responsible for the aphotrotropic phenotype of plants without CK2 activity. This internalization is, however, independent of the presence of NPH3 and therefore independent of ubiquitination. Surprisingly, internalization of phot1 observed in light conditions is also independent of the presence of NPH3.
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Leroy, Didier. "Interaction polyamines/protéine-kinase CK2 : étude moléculaire et fonctionnelle." Grenoble 1, 1996. http://www.theses.fr/1996GRE10046.
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La proteine-kinase ck2 est une serine/threonine proteine-kinase ubiquiste dans le monde eucaryote. Elle se compose de deux sous-unites catalytiques differentes alpha et alpha' et d'une sous-unite regulatrice beta qui s'assemblent sous la forme d'heterotetrameres. La ck2 utilise l'atp comme le gtp, phosphoryle des proteines et peptides substrats acides et necessite une concentration extraphysiologique d'ions magnesiums pour pouvoir exhiber son activite catalytique maximale. Bien que plusieurs evidences suggerent qu'in vivo, la ck2 joue un role indispensable, aucun mode de regulation cellulaire n'a pu etre propose pour cette kinase a l'heure actuelle. Parmi les candidats susceptibles de porter une telle fonction regulatrice, les polyamines sont des composes vis-a-vis desquels de nombreuses etudes ont ete realisees. Presentes en concentrations submillimolaires, ces molecules sont capables de stimuler l'activite catalytique de la ck2 d'un facteur 10 a 20 dans des conditions de concentrations physiologiques d'ions magnesiums. Lors de ces travaux, les parametres cinetiques regissant l'activation de la ck2 en presence de polyamines, ont ete definis. L'effet de la spermine sur la ck2 a ete analyse par dichroisme circulaire. L'interaction spermine-ck2 a ete caracterisee en presence de molecules analogues de la spermine ainsi qu'en ce qui concerne le type et le nombre de liaisons mises en jeu. L'utilisation d'un analogue photoactivable de la spermine a permis d'identifier un site majeur de liaison des polyamines sur la sous-unite beta au niveau du peptide t72-m78. Deux autres sites ont ete detectes au niveau des residus h108 de cette sous-unite et l220 de la sous-unite alpha. La construction, l'expression et la purification de proteines de fusion impliquant le domaine d51-p110 de la sous-unite beta ou la sous-unite beta entiere ont permis de confirmer les resultats du marquage de photoaffinite d'une part et de definir ce domaine comme autonome et fonctionnel d'autre part. Son utilisation a conduit entre autre a la mise en evidence des domaines impliques dans l'interaction des proteines fgf-2 et ck2. Aujourd'hui, la poursuite de ce travail par une approche cellulaire pourrait permettre d'apporter des elements de reponse quant a l'existence d'un role de regulateurs physiologiques joue par les polyamines vis-a-vis de la ck2
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Bosc, Denis G. "The catalytic subunits of protein kinase CK2, expression, covalent modification, and regulatory interactions." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1999. http://www.collectionscanada.ca/obj/s4/f2/dsk2/ftp03/NQ35040.pdf.
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Prudent, Renaud. "Identification et caractérisation d’inhibiteur de la protéine-kinase CK2." Grenoble 1, 2009. http://www.theses.fr/2009GRE10260.
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De nombreuses observations établissent un lien entre la dérégulation de la protéine kinase CK2 et certains cancers. Une activité élevée de CK2 dans certains cancers est un marqueur de mauvais pronostique. CK2 favorise la progression tumorale en régulant de nombreux oncogènes et suppresseur de tumeurs, ainsi qu’en protégeant des protéines anti-apoptotiques du clivage par les caspases. En conséquence, CK2 a émergé comme une cible thérapeutique et son inhibition pharmacologique représente une stratégie prometteuse. A l’instar d’autres protéine-kinases, des inhibiteurs ATP-compétitifs de CK2 ont déjà été identifié. Cependant, leur efficacité est variable, ce qui a conduit au développement de stratégies alternatives pour inhiber cette enzyme multi-protéique. Par criblage de chimiothèques à l’aide de CK2a recombinante, j’ai identifié des composés ciblant le site de fixation de l’ATP de la kinase ou un exosite. Ces composés ont été caractérisé structuralement par radiocristallographie aux rayons X ou par diffusion de rayons X aux petits angles (SAXS). Ces composés ont également été testés dans un test rapporteur de l’activité CK2 cellulaire. Certains inhibiteurs sont actifs sur CK2 dans des cellules vivantes. Deux composés (un ATP-compétitif et un allostérique) démontrent une activité anti-tumorale dans des tests de régression de xénogreffes tumorales dans des modèles murins. Ainsi, ces travaux ont conduit à l’identification des premiers inhibiteurs allostériques de CK2. Ceci montre que cibler CK2 hors du site de fixation de l’ATP est une alternative viable qui permettra d’exploiter de nouveaux mécanismes d’action et d’envisager de nouvelles opportunités thérapeutiquesExperimental evidence supports the view that disregulated Protein kinase CK2 is linked to cancers. Elevated CK2 activity in human cancer is an unfavorable prognostic marker. CK2 enhances progression of oncogenesis by regulating various oncogenes, tumor suppressor proteins and protecting anti-apoptotic proteins from caspase-mediated cleavage. Consequently, CK2 has emerged as a therapeutic target and its pharmacological inhibition appears as a promising strategy. Similar to other protein kinases, numerous ATP competitive inhibitors have been identified. However, they display variable effectiveness. Recently, alternative strategies to inhibit this multi-subunit enzyme have been revealed. Screening of chemical libraries using recombinant CK2a could identify compounds that target either the ATP binding site or exosites. These compounds were structurally characterized by analyzing CK2a-inhibitor complexes by means of X-ray structure crystallography or Small-Angle X-ray Scattering (SAXS). These compounds were also evaluated in a novel CK2 cellular activity assay. Several chemically unrelated inhibitors were found to be potent CK2 inhibitor in living cells. Two compounds (ATP-competitive and allosteric, respectively) showed anti-tumor activity, when tested in murine tumor xenograft regression assays. Taken together, this work leads to the identification of the first allosteric inhibitors of CK2, highlighting a new mode of inhibition of CK2. It also demonstrates that targeting an exosite on CK2 is a viable alternative to ATP-competitive inhibitors. This promises new opportunities by exploiting these new mechanisms of action
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Lebrin, Franck. "Implication de la protéine kinase CK2 dans la prolifération cellulaire." Université Joseph Fourier (Grenoble), 2000. http://www.theses.fr/2000GRE10164.
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La proteine kinase ck2 est une serine/threonine kinase qui se presente classiquement apres purification sous la forme d'un heterotetramere, compose de deux sous-unites catalytiques ck2alpha et/ou ck2alpha' associees fortement a deux sous-unites dites regulatrices ck2beta. Il s'agit d'une enzyme ubiquiste et phylogenetiquement conservee de la levure a l'homme dont la regulation et la fonction biologique sont encore meconnues. L'objectif de ce travail a ete de determiner la place et le role eventuel de la sous-unite catalytique ck2alpha dans la signalisation mitogenique et d'etudier la fonction de la proteine kinase ck2 dans la proliferation cellulaire des lignees de fibroblastes nih3t3 et ccl39. Au cours de cette etude, j'ai pu montrer que ck2alpha forme un complexe moleculaire avec la proteine phosphatase 2a (pp2a) et cible l'activite phosphatase sur mek1, composant central de la voie erk/mapk, connue pour reguler la proliferation cellulaire. La surexpression de ck2alpha est capable d'inhiber l'activation de mek1 et de erk/mapk par le serum. J'ai egalement pu montrer que ce mecanisme de regulation etait specifique du module erk/mapk. L'etude des effets biologiques de ce complexe ck2alpha-pp2a indique que la surexpression de ck2alpha inhibe la transformation cellulaire des fibroblastes nih3t3 induite par ras oncogenique, mais n'affecte pas la proliferation cellulaire. La surexpression des differentes formes de ck2 n'a aucun effet detectable sur la proliferation cellulaire. Par contre, l'utilisation d'un mutant dominant inactif de la sous-unite catalytique ck2alpha : ck2alphak68a inhibe la progression en g1 du cycle cellulaire des fibroblastes nih3t3 et ccl39. J'ai ensuite analyse le mecanisme d'action de ce mutant dominant inactif. J'ai pu montrer que ck2alphak68a s'associe a la sous-unite regulatrice ck2beta endogene et qu'il est incapable, ni de moduler l'expression des sous-unites de ck2 endogenes, ni de generer de ck2alpha libre detectable. J'ai montre que l'effet observe sur la proliferation ne depend pas de la capture par ck2alphak68a d'une population de ck2beta libre presente dans la cellule. Le mecanisme d'action de ck2alphak68a semble plutot reposer sur l'inhibition de la phosphorylation de substrats endogenes. Ces derniers restent a caracteriser.
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Bestgen, Benoit [Verfasser], and Rolf [Akademischer Betreuer] Hartmann. "Selective modulation of the protein kinase CK2 : discovery, syntheses and characterizationof non-ATP site inhibitors of CK2 / Benoit Bestgen. Betreuer: Rolf Hartmann." Saarbrücken : Saarländische Universitäts- und Landesbibliothek, 2016. http://d-nb.info/109622075X/34.
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Mitchell, Sophie Lousie. "A fragment-based drug discovery approach for the development of selective inhibitors of protein kinase CK2." Thesis, University of Cambridge, 2018. https://www.repository.cam.ac.uk/handle/1810/278650.
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Over the last twenty years, fragment-based drug discovery (FBDD) has emerged as a highly successful way to provide lead compounds for subsequent optimisation into drug candidates. Initial hits usually exhibit lower potency than those identified by more traditional techniques, such as High-Throughput Screening (HTS), but the optimisation phase of FBDD is highly efficient, thus providing superior lead-like compounds. The recent application of FBDD in a variety of protein kinase campaigns has successfully led to the identification of novel binding sites and highly efficient chemical ligands. This demonstrates the utility of the FBDD strategy against well-established kinase targets, where selectivity is otherwise challenging due to significant conservation of the ATP-binding site. Protein kinase CK2 is a ubiquitously expressed and constitutively active regulator of cell growth, proliferation and apoptosis. Elevated levels of CK2 protein and activity have historically been involved in human cancer, including lung, cervical and head and neck cancer types, and its overexpression is associated with worse prognosis. A number of CK2 inhibitors are currently available displaying activity against multiple cancers in vitro and in the clinic, however the majority of these candidates target the ATP-binding site and thus display poor selectivity in kinase panel assays. Here we explore the application of FBDD towards the development of potent and selective inhibitors of the catalytic α-subunit of CK2. This project exploits a novel, conserved binding site, named the αD pocket, for the generation of allosteric inhibitor molecules. Following structure-based optimisation of a potent inhibitor series, and characterisation of a previously unreported binding mode, a fragment linking strategy between the lead αD-site fragment and a low-affinity pseudosubstrate peptide is investigated. This work validates the utility of FBDD towards the discovery of new binding modes, presents a first in class CK2α allosteric inhibitor series and provides the first X-ray crystal structure of protein kinase CK2 in complex with a ligand binding in the substrate-binding channel.
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Trott, Regina L. "Drosophila melanogaster protein kinase CK2 interacts with and phosphorylates the neurogenic repressor m8 resulting in the production of a novel eye phenotype." Morgantown, W. Va. : [West Virginia University Libraries], 2005. https://eidr.wvu.edu/etd/documentdata.eTD?documentid=4407.
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Thesis (M.S.)--West Virginia University, 2005.Title from document title page. Document formatted into pages; contains vi, 90 p. : ill. (some col.). Includes abstract. Includes bibliographical references (p. 79-90).
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Lau, Faye. "Muscular activity regulates the expression of ColQ subunit of acetylcholinesterase : a signaling pathway mediated by Ca2̳+̳/ calmodulin-dependent protein kinase II /." View abstract or full-text, 2007. http://library.ust.hk/cgi/db/thesis.pl?BIOL%202007%20LAU.
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Laudet, Béatrice. "Stratégies pour inhiber une interaction protéine-protéine de haute affinité : l'exemple de la protéine kinase CK2." Grenoble 1, 2007. http://www.theses.fr/2007GRE10172.
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Les nombreux arguments qui plaident en faveur du potentiel oncogénique de la protéine kinase CK2 en font une cible thérapeutique prometteuse en cancérologie. Cette protéine kinase se présente sous la forme d'un complexe de deux sous-unités catalytiques CK2α constitutivement actives et d'un dimère de sous-unités régulatrices CK2ß. Notre laboratoire a montré que l'interaction dynamique entre ces sous-unités dans la cellule est une composante essentielle dans la régulation de cette enzyme. Pour mieux comprendre cette régulation dans les processus cellulaires normaux et pathologiques, il apparaît nécessaire de développer des molécules capables de perturber cette interaction protéine-protéine. Pour cela, trois stratégies complémentaires ont été utilisées : 1) caractérisation des « hot spots » de l'interaction CK2α- CK2ß à partir de la structure cristallographique du tétramère ; 2) conception rationnelle du premier ligand antagoniste de cette interaction sous la forme d'un peptide cyclique mimétique (IC50 = 3 μM) ; 3) définition à partir de ce peptide d'un pharmacophore utilisable pour identifier des molécules chimiques analogues par une approche de criblage virtuel. Un cluster de molécules inhibitrices actives in vitro et in vivo a ainsi été identifié. Elles représentent les premiers inhibiteurs chimiques de cette interactionMany arguments in favour of oncogenic potential of CK2 protein kinase make it a promising therapeutic target in oncology. This protein kinase is composed of a tetrameric complex of two catalytic subunits CK2a constitutively active and a dimmer of two regulatory subunits CK2b. Our laboratory showed that dynamic interaction between these two subunits in cell is an essential component for this enzyme regulation. For better understanding this regulation in normal and pathologic processes, it seems necessary to develop compounds able to perturb this proteinprotein interaction. In this respect, three complementary strategies were used: 1) hot spots characterization for CK2a-CK2b interaction based on tetramer crystal structure. 2) rational conception of the first antagonist of this interaction as a mimetic cyclic peptide (IC50 = 3 mM). 3) pharmacophore definition based on this peptide allowing to identify chemical molecules analogs by virtual screening. A cluster of chemical compounds active as well in vitro as in vivo has been identified. They represent the first inhibitors for this interaction
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Alcaraz, Muñoz Estefania. "The role of Protein Kinase CK2 in pro-survival pathways in clear cell renal cell carcinoma (ccRCC) cells." Doctoral thesis, Universitat Autònoma de Barcelona, 2016. http://hdl.handle.net/10803/381067.
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La proteïna quinasa CK2 és una Serina/Treonina quinasa àmpliament expressada en tots els organismes eucariotes. Fins el dia d’avui, més de 300 substrats per aquesta quinasa han estat descoberts, essent molts d’ells essencials per la viabilitat cel·lular. Per aquest motiu, CK2 ha estat considerada com una quinasa decisiva per la viabilitat de totes les cèl·lules eucariotes.
CK2 juga un paper clau en la supervivència cel·lular, proliferació i mecanismes antiapoptòtics, observant-se que la seva desregulació es troba associada a diferents malignitats humanes. A més a més, els efectes pleiotròpics d’aquesta proteïna han connectat CK2 amb altres vies que són crucials en diferents processos involucrats en la tumorigenesis. Malgrat tot, poc es coneix sobre la implicació de CK2 i les vies de senyalització en el carcinoma renal de cèl·lules clares (ccRCC).
En aquest treball s’ha volgut estudiar la implicació de CK2 en les bases moleculars del ccRCC, analitzant l’efecte de la inhibició de CK2 sobre les vies de senyalització de Akt i ERK1/2 davant la resposta a heparin-binding EGF-like growth factor (HB-EGF), així com la connexió entre CK2 i ErbB4, proteïnes que s’han trobat disminuïdes en ccRCC. Per tal d’assolir aquest objectiu, l’activitat de CK2 ha estat inhibida mitjançant l’ús de diferents inhibidors farmacològics, i les subunitats reguladores i catalítiques de CK2 han estat silenciades de forma independent mitjançant short hairpin RNA (shRNA) en cèl·lules de túbul proximal derivades d’un ronyó sa (HK-2) i cèl·lules derivades d’un adenocarcinoma primari de cèl·lules clares (786-O).
El resultat més impactant és que la inhibició de CK2, tant per inhibidors químics com per shRNA, perjudica l’activació de Akt i ERK1/2 en resposta a HB-EGF, que són vies de senyalització decisives involucrades en el procés de mort cel·lular, proliferació i autofàgia. De la mateixa forma, la disminució de la subunitat reguladora CK2β ve acompanyada de canvis en l’expressió de marcadors de transició epiteli-mesènquima (EMT), tals com E-caderina i Snail1. És interessant assenyalar que els resultats d’aquests estudi suggereixen que la disminució de CK2β indueixen l’expressió de HIF-α i la fosforilació de STAT3, contribuint a la regulació de E-caderina i Snail1 en les cèl·lules derivades de ccRCC. Per altra banda, la sobre-expressió de ErbB4 en 786-O altera la proliferació cel·lular, així com potencia l’activació de Akt i ERK1/2 davant de l’estimulació amb HB-EGF. A més a més, la inhibició de CK2 per CX-4945 i el silenciament de CK2β per siRNAs causa una reducció significativa dels nivells de ErbB4. Els resultats d’aquesta cerca mostren que CK2 afecta a components claus de les vies de senyalització, tals com ErbB4, Akt, ERK1/2, HIF-α, Snail 1 i STAT3 en cèl·lules renals, recolzant la implicació de CK2 en el ccRCC.Protein kinase CK2 is a Serine/Threonine kinase widely expressed in all eukaryotic organisms. To date, more than 300 substrates for this kinase have been discovered, most of them essential for cell viability. For that reason, CK2 has been considered as a protein kinase decisive for the viability of all eukaryotic cells. CK2 plays pivotal roles in cell survival, proliferation and anti-apoptotic mechanisms, and its dysregulation is associated with human malignancies. Furthermore, the pleiotropic effects of this protein have connected CK2 with other pathways that are crucial in several processes involved in tumorigenesis. However, little is known on the potential cross-talk between CK2 and these signalling pathways in clear cell renal cell carcinoma (ccRCC) cells. The purpose of this work has been to study the involvement of CK2 in the molecular basis of ccRCC, analysing the effect of CK2 inhibition on Akt and ERK1/2 signalling pathways in response to heparin-binding EGF-like growth factor (HB-EGF), as well as the connection between CK2 and ErbB4, which has been found downregulated in ccRCC. In order to assess this gkoal, CK2 activity has been targeted by pharmacological inhibitors, and the regulatory and catalytic CK2 subunits have been independently silenced by short hairpin RNA (shRNA), in tubular proximal cells derived from normal kidney (HK-2), and cells derived from a primary clear cell adenocarcinoma (786-O). The most striking result is that CK2 inhibition, either by chemical inhibitors or shRNA downregulation, impairs the activation of Akt and ERK1/2 in response to HB-EGF, which are decisive signalling pathways involved in the process of cell death, proliferation and autophagy. Likewise, downregulation of regulatory subunit CK2β is accompanied by changes in the expression of Epithelial-Mesenchymal-Transition (EMT) markers, such as E-cadherin and Snail1. Interestingly, the results of this study suggest that CK2β downregulation induces HIF-α expression and STAT3 phosphorylation which may contribute to E-cadherin and Snail1 regulation in ccRCC cells. On the other hand, overexpression of ErbB4 in 786-O cells alters cell proliferation as well as enhances HB-EGF-induced Akt and ERK1/2 activation. In addition, CK2 inhibition by CX-4945 and downregulation of CK2β by siRNAs results in a significant reduction of ErbB4 levels.The results of this research show that CK2 affects key components of signalling pathways, such as ErbB4, Akt, ERK1/2, HIF-α, Snail 1 and STAT3 in renal cells, supporting the potential involvement of CK2 in ccRCC.
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Ulges, Alexander [Verfasser]. "Funktionelle Charakterisierung der Protein-Kinase CK2 in der Suppression Th2-vermittelter Immunantworten durch regulatorische T-Zellen / Alexander Ulges." Mainz : Universitätsbibliothek Mainz, 2014. http://d-nb.info/1058996835/34.
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Liu, Jin. "Increased CKIP-1 suppresses Smad-dependent BMP signaling to inhibit bone formation during aging." HKBU Institutional Repository, 2016. https://repository.hkbu.edu.hk/etd_oa/327.
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Emerging evidence indicates that the dysregulation of protein ubiquitination plays a crucial role in aging-associated diseases. Smad-dependent canonical BMP signaling pathway is indispensable for osteoblastic bone formation, which could be disrupted by the ubiquitination and subsequent proteasomal degradation of Smad1/5, the key molecules for BMP signaling transduction. However, whether the dysregulation of Smad1/5 ubiquitination and disrupted BMP signaling pathway are responsible for the age-related bone formation reduction is still underexplored. Casein kinase-2 interacting protein-1 (CKIP-1), also known as Pleckstrin homology domain-containing family O member 1 (PLEKHO1), is a previously identified ubiquitination-related molecule that could specifically target the linker region between the WW domains of Smurf1 to promote the ubiquitination of Smad1/5. Here, we found an age-related increase in the expression of CKIP-1 in bone specimens from either fractured patients or aging rodents, which was associated with the age-related reduction in Smad-dependent BMP signaling and bone formation. By genetic approach, we demonstrated that loss of Ckip-1 in osteoblasts could promote the Smad-dependent BMP signaling and alleviated the age-related bone formation reduction. In addition, osteoblast-specific Smad1 overexpression had beneficial effect on bone formation during aging, which could be counteracted after overexpressing Ckip-1 within osteoblasts. By pharmacological approach, we showed that osteoblast-targeted CKIP-1 siRNA treatment could enhance Smad-dependent BMP signaling and promote bone formation in aging rodents. Taken together, it suggests that the increased CKIP-1 could suppress Smad-dependent BMP signaling to inhibit bone formation during aging, indicating the translational potential of targeting CKIP-1 in osteoblast as a novel bone anabolic strategy for reversing established osteoporosis during aging.
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Mackenbach, Loue Petra. "Translocation nucléaire de la protéine kinase CK2 induite par les facteurs de croissance et surexpression d'une forme anormale de la kinase dans les tumeurs du sein." Université Joseph Fourier (Grenoble ; 1971-2015), 1997. http://www.theses.fr/1997GRE10164.
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La proteine kinase ck2 est une serine/threonine proteine kinase exprimee dans toutes les cellules eucaryotes. Elle est composee de deux sous-unites catalytiques (alpha et alpha') et deux sous-unites regulatrices beta. Des observations convergentes suggerent qu'elle doit jouer un role important dans le controle de la division cellulaire. Le but de notre travail etait de tenter de determiner la facon dont les facteurs de croissance peuvent reguler la ck2 dans la cellule. Dans un premier temps nous avons caracterise les differentes isoformes de la ck2 mais aucune distinction n'a ete detectee concernant leur distribution dans la cellule et leurs activites catalytiques respectives. Par des etudes en immunofluorescence, nous avons pu mettre en evidence que la translocation nucleaire de la ck2 est dependante de l'activite tyrosine kinase du recepteur a l'egf et de l'activite de ras et concerne donc avant tout la reponse cellulaire a une signalisation mitogene. La biosynthese de la ck2 semble egalement etre sensible aux facteurs de croissance. En effet, des la premiere heure de stimulation, une augmentation de biosynthese est detectable. L'ensemble de ces resultats montre que les facteurs de croissance regulent a la fois la biosynthese et la localisation de la ck2 dans la cellule. La poursuite de cette etude par une approche de biologie moleculaire pourrait permettre de determiner l'importance des differents domaines des sous-unites de la ck2 dans le controle de la localisation subcellulaire de la kinase. Dans une deuxieme partie nous avons etudie la ck2 dans les tumeurs du sein. Ce tissu exprime six fois plus d'activite ck2 que le tissu normal et cette situation est correlee avec une surexpression de la sous-unite catalytique. De plus, une sous-unite regulatrice anormale, associee a la ck2 alpha a ete mise en evidence : la proteine est deletee de ses deux derniers acides amines c-terminaux. L'existence de cette forme de ck2 dans les coupes de tumeurs nous permet de conclure que la presence de cette sous-unite tronquee est une realite dans les tumeurs. Le defi est maintenant de poursuivre de telles recherches par une approche cellulaire afin de determiner la relation entre la proteolyse de la ck2 beta et la tumorigenese. La quantification de la ck2 beta anormale dans les tumeurs devrait nous permettre de determiner si cette proteine est un nouveau marqueur tumoral.
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Silva, Simone Sandy da. "Influência da ck2 no processo de interação Leishmania donovani-macrófagos." Universidade do Estado do Rio de Janeiro, 2012. http://www.bdtd.uerj.br/tde_busca/arquivo.php?codArquivo=7180.
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O gênero Leishmania apresenta espécies capazes de desenvolver doenças de grande importância para a saúde pública, as leishmanioses, que apresentam prevalência mundial de 12 milhões de pessoas. Quando os parasitos entram em contato com o hospedeiro humano passam por um processo de metaciclogênese adquirindo capacidade de interagir com os macrófagos. Inúmeras atividades biológicas são desencadeadas pela ativação de sistemas de transdução de sinais, onde as proteínas cinases e fosfatases desempenham papel fundamental. A proteína cinase CK2 parece estar presente em todas as células eucarióticas (núcleo, citoplasma e superfície). É caracterizada como enzima serina/treonina cinase, embora também seja capaz de fosforilar resíduos de tirosina em suas proteínas-alvo. No presente trabalho, demonstramos que o principal inibidor da CK2, TBB, foi capaz de inibir o crescimento de formas promastigotas de L. donovani e mostrou um mecanismo de ação irreversível, entretanto não foi capaz de induzir apoptose nas formas promastigotas de L. donovani. O pré-tratamento dos parasitos e macrófagos, assim como a adição do TBB durante o processo de infecção induziram uma redução significativa no número de amastigotas por macrófagos possivelmente pelo mecanismo de morte celular programada demosntrada pela técnica do TUNEL. O tratamento de macrófagos com TBB não induziram o aumento de óxido nítrico. Ensaios de imunofluorescência demonstraram a presença de CK2α em promastigotas. Macrófagos não infectados demonstraram pouca marcação para CK2α. Após a interação, a enzima mostrou-se distribuída preferencialmente na periferia dos macrófagos. Os dados do trabalho sugerem que a CK2 é uma importante enzima para a atividade biológica da Leishmania donovani, tendo seu estudo importante relevância para a descoberta de novos alvos terapêuticos.The genus Leishmania contains multiple species that are responsible for a group of diseases, leishmaniasis. Current estimates suggest that 12 million people are infected. When parasites enter a human host, they undergo metacyclogenesis and acquire the ability to interact with macrophages. This interaction activites signal transduction pathways inducing numerous biological activities, including protein kinase CK2. CK2 has been observed in all eukaryotic cells residing in the nucleus, the cytoplasm and on the cell surface. It appears to have an essential function and recognizes serine/threonine or tyrosine residues in target proteins for phosphorylation. In the present study, we demonstrate that the treatment of L. donovani promastigotes with TBB resulted in inhibition of cell growth and showed an irreversible mechanism of action, however was not able to induce apoptosis in L. donovani promastigotes. The pre-treatment of promastigotes and macrophages, as well as the addition of TBB during infection induced a significant reduction in the number of amastigotes per macrophages, possibly via the mechanism of programmed cell death showed by TUNEL technique. Treatment of macrophages with TBB did not induce the increase of nitric oxide. Immunofluorescence assays demonstrated the presence of CK2α distributed throughout the surface the promastigotes. Uninfected macrophages showed little to no CK2α. After initiating the interaction of parasites with macrophages, CK2α was observed distributed preferentially in the nucleus of macrophages. Job data suggest that CK2 is an important enzyme for the biological activity of Leishmania donovani, and its study important relevance to the discovery of new therapeutic targets.
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Stark, Felix [Verfasser], and Thomas [Akademischer Betreuer] Raabe. "Funktionelle Untersuchungen zur Regulation der Protein Kinase CK2 durch Polyamine in Drosophila melanogaster und deren physiologische Bedeutung / Felix Stark. Betreuer: Thomas Raabe." Würzburg : Universitätsbibliothek der Universität Würzburg, 2011. http://d-nb.info/1013620003/34.
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Caldarella, Eleonora. "Functional implications of protein kinase CK2 in F-actin dynamics and in the cross-talk between auxin and salicylic acid signalling pathways in arabidopsis thaliana." Doctoral thesis, Universitat Autònoma de Barcelona, 2018. http://hdl.handle.net/10803/565670.
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El objetivo de esta tesis era el estudio del papel de la proteina CK2 en tres
procesos biológicos de plantas, utilizando Arabidopsis thaliana como modelo:
fototropismos, regulación de las vías de señalización del ácido salicílico y de las auxinas,
y dinámica del citoesqueleto de actina. Un mutante de pérdida de función de CK2
(CK2mut) obtenido por otros miembros del laboratorio, que mostró rasgos fenotípicos
relacionados con alteraciones en procesos dependientes de auxinas, ha sido la
herramienta principal utilizada en esta tesis. Tras el tratamiento con luz azul, el mutante
mostró una pérdida de la respuesta fototrópica en hipocotilos y un aumento de la
curvatura fototrópica de la raíz. Por otra parte, la internalización de PIN2 en
compartimentos vacuolares, que normalmente ocurre cuando las plántulas se transfieren
a oscuridad, estaba ausente en las plántulas CK2mut, aunque el reciclaje de PIN2 a la
membrana plasmatica no se veía afectado.
Para comprender mejor la regulación de la señalización por auxinas en plantas
defectivas en CK2 estudiamos la estabilidad de las proteínas AUX/IAA, que son
represoras de las vías de señalización de auxinas y modulan las respuestas de auxinas.
Demostramos que la inhibición de CK2 dio como resultado un aumento de la estabilidad
del represor AXR3. Como las plántulas CK2mut contienen niveles de ácido salicílico (SA)
anormalmente elevados, estudiamos también la regulación cruzada (crosstalk) entre la
señalización por auxina y por SA, y demostramos que el aumento de estabilidad de AXR3
estaba mediada por SA.
Los estudios in vivo por microscopía confocal de la arquitectura de actina en
células de la raíz se realizaron utilizando el reporter de actina GFP-FABD2. Las células
del mutante de CK2 mostraron una fuerte desorganización de la estructura reticular y un
colapso de los haces de F-actina. Se realizaron estudios de turnover de actina que
confirmaron que la falta de actividad de CK2 afectaba fuertemente a la polimerización de
los filamentos de actina. El turnover de F-actina está regulado por proteínas de unión a
actina, entre ellas la familia de los factores de despolimerización de actina (ADF).
Utilizando herramientas de bioinformática, encontramos algunos sitios putativos de
fosforilación para CK2 en la secuencia aminoacídica de ADF4. Ello podría indicar que la
CK2 podría regular la actividad de ADF4, involucrada en el desensamblaje del filamento
de actina.The aim of this thesis was the study of the role of protein kinase CK2 in three plant biological processes, using Arabidopsis thaliana as a model: plant phototropisms, regulation of salicylic acid and auxin signalling pathways, and dynamic of actin cytoskeleton. A loss-of-function mutant of CK2 (CK2mut), obtained by former members of the laboratory and that showed phenotype traits linked to alterations in auxin dependent processes, was used as the main tool for our studies. In the present work we show that CK2mut seedlings exhibit loss of the hypocotyl phototropic response and increase of the root phototropic curvature under blue light treatments. Moreover, PIN2 internalization into vacuolar compartments after transferring CK2mut seedlings to darkness did not occur when CK2 activity was depleted, although PIN2 recycling to the plasma membrane was not affected. In order to better understand auxin-signalling regulation in CK2-defective plants we studied the stability of AUX/IAA proteins, which are repressors of auxin signalling pathways and modulate responses to auxin. We performed histochemical and fluorimetric assays and we demonstrated that inhibition of CK2 resulted in over-stabilization of the AXR3 repressor. As CK2mut seedlings contain high levels of salicylic acid (SA), we then studied the cross-talk between auxin and SA pathways and we demonstrated that the over-stabilization of AXR3 in CK2- defective plants is mediated by SA. In vivo confocal imaging of actin architecture was performed by using the actin reporter GFP-FABD2. CK2-depleted seedlings showed strong disorganization of the actin network and collapse of F-actin bundles. Studies of actin turnover in Arabidopsis seedlings confirmed that the lack of CK2 activity strongly affects polymerisation of actin filaments. F-actin turnover is regulated by actin binding proteins, among them the family of actin depolymerisation factors (ADFs). In our study, we found some putative predicted phosphorylation sites for CK2 in ADF4. Thus, we can speculate that CK2 might be involved in the regulation of the ADF4 actin filament disassembly activity.
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Alchab, Faten. "Synthèse et évaluation de dérivés de l'indéno[1,2-b]indole comme inhibiteurs potentiels de la protéine kinase humaine CK2." Thesis, Lyon 1, 2013. http://www.theses.fr/2013LYO10162.
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La protéine kinase caséine kinase 2 (CK2) est une sérine/thréonine kinase hautement pléiotrope dont la liste des substrats est supérieure à 500 protéines, lesquelles sont impliquées dans un large éventail de fonctions cellulaires. Les sous-unités catalytiques de CK2 (alpha et/ou alpha') sont constitutivement actives soit seules soit en combinaison avec les sous-unités régulatrices béta pour former une protéine hétérotétramérique (holoenzyme). Une troisième isoforme de la sous-unité catalytique, désignée CK2α'', a été découverte plus récemment et peu d'informations sont actuellement disponibles. L'activité hautement constitutive de CK2 est suspectée de contribuer au phénomène de néoplasie. Une stratégie de conception d'inhibiteurs tétracycliques ciblant le site ATP de la CK2 a permis l'élaboration de trois séries de composés comportant le motif indéno[1,2-b]indole. Un procédé multi-étapes de synthèse a permis de fonctionnaliser précisément le cycle D du noyau indéno[1,2-b]indole et de générer une première chimiothèque de molécules originales. Toutes les molécules finales ont été testées sur la protéine kinase humaine CK2 (Muenster) et certaines ont présentées des CI50 de l'ordre du submicromolaire. L'analyse des Relations Structure-Activité (SAR) et la construction d'un modèle 3D-QSAR (Duesseldorf) a contribué à affiner le choix des substituants introduits sur le châssis moléculaire développé. Les indéno[1,2-b]indoles fonctionnalisés les plus prometteurs ont été également testés sur d'autres cibles biologiques comme la phosphatase CDC25A (Metz) et la kinase DYRK1B (Saarbruecken). Des études de modélisation moléculaire (Duesseldorf) utilisant les données cristallographiques disponibles de l'enzyme ont permis d'analyser les interactions ligand-protéine. Les inhibiteurs les plus puissants in vitro ont été testés sur quatre lignées cellulaires normales afin d'établir leur profil cytotoxique (Centre de Recherche en Cancérologie de Lyon)Synthesis and evaluation of indéno[1,2-b]indole derivatives as potential inhibitors of human protein kinase CK2 Protein kinase casein kinase 2 (CK2) is a serine/threonine kinase highly pleiotropic listed substrates it is greater than 500 proteins, which are involved in a wide range of cellular functions. The catalytic subunits of CK2 (α and/or α') are constitutively active either alone or in combination with the regulatory subunits to form a hetero- beta protein holoenzyme). A third isoform of the catalytic subunit, designated CK2 α', was discovered more recently and little information is currently available. The high constitutive activity of CK2 is suspected of contributing to the phenomenal of neoplasia. A design strategy tetracyclic inhibitors targeting the ATP site of CK2 resulted in the development of three series of compounds containing the motif indeno[1,2-b]indole. A multi-step synthesis process has specifically functionalize the D ring of the core indeno[1,2-b]indole and generate a first combinatorial library of original molecules. All final compounds were tested on human protein kinase CK2 (Muenster), and some have reported IC50 of the order of sub-micromolar. Analysis of Structure-Activity Relationships (SAR) and the construction of a 3D-QSAR model (Duesseldorf) helped to refine the choice of substituents introduced into the moleculair frame developed. The indeno[1,2-b]indole the most promising functionalized indoles were also tested on other biological targets such as phosphatase CDC25 A (Metz) and kinase DYRK1B (Saarbruecken). Of molecular modeling studies (Duesseldorf) using the crystallographic data of the enzyme were used to analyze protein-ligand interactions. The most potent in vitro inhibitor were tested on four normal cell lines to determine their cytotoxic profile (Cancer Research Center of Lyon)
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Valero, Emmanuelle. "Mise en évidence, caractérisation, et étude des relations structure-fonction des différentes formes moléculaires de la protéine kinase CK2." Grenoble 1, 1996. http://www.theses.fr/1996GRE10216.
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La proteine kinase ck2 est une serine/threonine kinase ubiquiste chez les eucaryotes. Historiquement, la structure heterotetramerique (alpha#2beta#2) de l'enzyme a ete determinee dans des conditions de haute force ionique, car a faible force ionique, la kinase s'agrege. J'ai montre qu'il ne s'agit pas d'une agregation mais d'un veritable phenomene de polymerisation reversible engendrant trois formes moleculaires differentes: le protomere, alpha#2beta#2, l'anneau, (alpha#2beta#2)4, le filament fin, alpha#2beta#2)n, le filament epais, (alpha#2beta#2)4)#n. La force ionique ainsi que la concentration de l'enzyme sont les deux parametres essentiels qui conditionnent la structure quaternaire de la pkck2. Or, dans les conditions de force ionique et de concentration en pkck2 physiologiques, l'enzyme se presente sous la forme d'un melange de protomeres et d'anneaux. Ce resultat remet en question la structure quaternaire native reelle de la pkck2 in vivo. L'activite de la kinase varie avec sa concentration, et donc avec son niveau de polymerisation: les differentes formes moleculaires portent donc des activites specifiques differentes. L'anneau est la structure quaternaire que l'enzyme adopte dans des conditions optimales de catalyse. De plus, les stimulateurs de l'activite catalytique de la kinase stabilisent la forme moleculaire anneau, alors que les inhibiteurs favorisent les structures protomeriques ou filamentaires. Ceci suggere fortement que l'anneau represente la forme active de la kinase, les autres formes moleculaires correspondant a une enzyme faiblement active. L'ensemble de ces travaux met donc en evidence une regulation possible de l'activite catalytique de la pkck2 via sa structure quaternaire. La proteine kinase ck2 doit donc etre consideree comme une enzyme dissociante, sa structure quaternaire est un nouveau parametre fondamental a prendre en compte lors de toute etude concernant cette proteine kinase
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Salmonowicz, Daniel J. "Creation of a Unique GST-FAK Plasmid for Protein Expression." Ohio Dominican University Honors Theses / OhioLINK, 2020. http://rave.ohiolink.edu/etdc/view?acc_num=oduhonors1588678705964411.
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Ciocea, Alieta. "A casein kinase 2 inhibitor is a potent anti-cancer drug candidate." Cleveland, Ohio : Cleveland State University, 2008. http://rave.ohiolink.edu/etdc/view?acc_num=csu1210258333.
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Thesis (Ph.D.)--Cleveland State University, 2008.Abstract. Title from PDF t.p. (viewed on Oct. 8, 2008). Includes bibliographical references. Available online via the OhioLINK ETD Center. Also available in print.
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Khan, Dilshad Hussain. "Role of histone deacetylases in gene expression and RNA splicing." Informa UK Limited, 2012. http://hdl.handle.net/1993/22163.
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Histone deacetylases (HDAC) 1 and 2 play crucial role in chromatin remodeling and gene expression regimes, as part of multiprotein corepressor complexes. Protein kinase CK2-driven phosphorylation of HDAC1 and 2 regulates their catalytic activities and is required to form the corepressor complexes. Phosphorylation-mediated differential distributions of HDAC1 and 2 complexes in regulatory and coding regions of transcribed genes catalyze the dynamic protein acetylation of histones and other proteins, thereby influence gene expression.
During mitosis, highly phosphorylated HDAC1 and 2 heterodimers dissociate and displace from mitotic chromosomes. Our goal was to identify the kinase involved in mitotic phosphorylation of HDAC1 and 2. We postulated that CK2-mediated increased phosphorylation of HDAC1 and 2 leads to dissociation of the heterodimers, and, the mitotic chromosomal exclusions of HDAC1 and 2 are largely due to the displacement of HDAC-associated proteins and transcription factors, which recruit HDACs, from chromosomes during mitosis. We further explored the role of un- or monomodified HDAC1 and 2 complexes in immediate-early genes (IEGs), FOSL1 (FOS-like antigen-1) and MCL1 (Myeloid cell leukemia-1), regulation. Dynamic histone acetylation is an important regulator of these genes that are overexpressed in a number of diseases and cancers. We hypothesized that transcription dependent recruitment of HDAC1 and 2 complexes over the gene body regions plays a regulatory role in transcription and splicing regulation of these genes.
We present evidence that CK2-catalyzed increased phosphorylation of HDAC1 and 2 regulates the formation of distinct corepressor complexes containing either HDAC1 or HDAC2 homodimers during mitosis, which might target cellular factors. Furthermore, the exclusion of HDAC-recruiting proteins is the major factor for their displacement from mitotic chromosomes. We further demonstrated that un- or monophosphorylated HDAC1 and 2 are associated with gene body of FOSL1 in a transcription dependent manner. However, HDAC inhibitors prevented FOSL1 activation independently of the nucleosome response pathway, which is required for IEG induction. Interestingly, our mass spectrometry results revealed that HDAC1 and 2 interact with a number of splicing proteins, in particular, with serine/arginine-rich splicing factor 1 (SRSF1). HDAC1 and 2 are co-occupied with SRSF1 over gene body regions of FOSL1 and MCL1, regardless of underlying splicing mechanisms. Using siRNA-mediated knockdown approaches and HDAC inhibitors, we demonstrated that alternative splicing of MCL1 is regulated by RNA-directed localized changes in the histone acetylation levels at the alternative exon. The change in histone acetylation levels correlates with the increased transcription elongation and results in change in MCL1 splicing by exon skipping mechanism.
Taken together, our results contribute to further understanding of how the multi-faceted HDAC1 and 2 complexes can be regulated and function in various processes, including, but not limited to, transcription regulation and alternative splicing. This can be an exciting area of future research for therapeutic interventions.
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Bourayne, Marie de. "Rôle de la CK2 dans l’activation de la réponse immunitaire induite par les molécules allergisantes et son lien avec Nrf2." Thesis, Paris 11, 2015. http://www.theses.fr/2015PA114823/document.
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L’eczéma allergique de contact (EAC) est une réaction inflammatoire aiguë médiée par les lymphocytes T (LT), survenant suite à l’exposition répétée de la peau avec une molécule allergisante présente dans l’environnement quotidien ou professionnel. Les molécules allergisantes sont des composés de faible poids moléculaire, appelés haptènes, qui activent les cellules dendritiques (DC). Les DC jouent un rôle essentiel dans la mise en place d’un EAC : elles acquièrent un phénotype mature, contrôlé par la voie des MAPK et la voie NF-B, leur permettant de présenter l’haptène aux LT afin d’initier ainsi une réponse immunitaire spécifique.Nous avons identifié au sein de la DC une nouvelle kinase, la protéine kinase CK2, indispensable à l’acquisition d’un phénotype mature et à la sécrétion de cytokines pro-inflammatoires clés dans l’orientation d’une réponse immunitaire. L’activité de la CK2 dans la DC est nécessaire à la génération d’une réponse Th1 en contrôlant la sécrétion d’IFN par les LT, et maintient une réponse Th17 préexistante. De plus, la CK2 permet à la DC de contrôler l’induction d’une réponse Th2 spontanée. Par ailleurs, la CK2 contrôle l’expression des gènes cibles de Nrf2, un facteur de transcription majeur dans la lutte contre le stress chimique induit par les haptènes. L’activation de Nrf2 met en jeu de nombreuses voies de signalisation, et nous avons mis en évidence c-Jun, facteur de transcription activé par les molécules allergisantes, comme un potentiel partenaire transcriptionnel de Nrf2Allergic contact dermatitis represents a severe health problem with increasing worldwide prevalence. It is a T cell-mediated inflammatory skin disease caused by chemicals present in daily or professional environment. Contact sensitizers are low molecular weight compounds termed haptens. These molecules are known to induce an up-regulation of phenotypic markers and cytokine secretion in dendritic cells (DCs), professional antigen-presenting cells, leading to the generation of effector T lymphocytes (LT).We identified a new kinase, termed CK2 (formerly casein kinase 2), as a key kinase in DCs in the acquisition of a mature phenotype and in the production of pro-inflammatory cytokines, involved in T cell polarization in response to contact sensitizers. CK2 activity in DC is necessary to induce a Th1 polarization by controlling the secretion of IFN- by LT, and maintains a pre-existing Th17 response. Moreover, CK2 in DC negatively controls a spontaneous Th2 response.Finally, CK2 controls the expression of Nrf2 target genes mRNA. Nrf2 is a protective transcription factor playing a major role in detoxification, oxidative stress and allergic inflammation generated by contact sensitizers. Nrf2 activation involves different kinases and we highlight that c-Jun could be bound to Nrf2 to generate an active transcriptional complex in response to chemicals
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Livecchi, Marion. "Synthèse pallado-catalysée de 5-azaindoles et évaluation de leur activité inhibitrice sur les protéines kinases CK2 et Pim-1." Thesis, Paris 5, 2013. http://www.theses.fr/2013PA05P616.
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L’inhibition de protéines kinases constitue une voie pleine de promesses pour la découverte de nouvelles thérapies ciblées contre le cancer. En 2003, le criblage de la chimiothèque de l’Institut Curie/CNRS a permis de mettre en évidence une famille de composés actifs sur deux de ces enzymes : CK2 et Pim-1. L’objectif de cette thèse était de synthétiser des analogues des « hits » de la chimiothèque possédant le squelette 5-azaindole afin d’en améliorer les propriétés biologiques. La préparation de tels composés étant peu décrite dans la littérature, trois voies de synthèse flexibles et efficaces ont été développées. L’élaboration de 5-azaindoles diarylés symétriques a tout d’abord été mise au point par hétéroannélation pallado-catalysée à partir de dérivés de la 4-aminopyridine. Les composés monosubstitués en position 2 ont ensuite été obtenus par réaction domino sila-Sonogashira/cyclisation 5-endo. Enfin, un procédé one-pot couplage de Sonogashira/aminopalladation/élimination réductrice a permis d’accéder aux molécules diarylées non symétriques avec une régiosélectivité contrôlée. L’application de ces méthodologies a conduit à la préparation de 70 composés fonctionnalisés dont la cytotoxicité et l’activité inhibitrice sur CK2 ont été évaluées. Une étude structure-activité a été réalisée et les fragments d’intérêt que doit posséder une molécule de type 5-azaindole pour inhiber efficacement la kinase ont ainsi été identifiésProtein kinases represent promising targets for anti-cancer drug design. In 2003, inhibitors of two of these enzymes, CK2 and Pim-1, were identified by the screening of the Curie Institute/CNRS small-molecule library. The aim of this thesis was to synthesize derivatives of these hits with a 5-azaindole scaffold in order to optimize their biological activity. As the synthesis of such molecules was not reported in the literature, efficient and flexible procedures were developed to access to these structures. Diarylated symmetrical 5-azaindoles were thus prepared by palladium-catalyzed heteroannulation from 4-aminopyridines derivatives. The methodology was subsequently extended to silylalkynes and led to monoarylated products through domino sila-Sonogashira/5-endo cyclization. Finally, a one-pot Sonogashira coupling/aminopalladation/reductive elimination afforded unsymmetrical compounds with a total control of the regioselectivity. Using these methodologies, 70 functionalized molecules were easily prepared. Their cytotoxicity and biological activity as CK2 inhibitors were then evaluated. A structure-activity relationship study was performed, which led to the identification of two key structural elements for the CK2 inhibitory potency of 5-azaindoles
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Aparicio, Siegmund Samadhi [Verfasser], Jürgen [Akademischer Betreuer] Scheller, and Lutz [Akademischer Betreuer] Schmitt. "Rezeptor cross-talk und die Rolle der Protein Kinase CK2 in der Signaltransduktion von Zytokinen der Interleukin (IL)-6/IL-12-Familie / Samadhi Aparicio Siegmund. Gutachter: Jürgen Scheller ; Lutz Schmitt." Düsseldorf : Universitäts- und Landesbibliothek der Heinrich-Heine-Universität Düsseldorf, 2015. http://d-nb.info/1067229175/34.
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Apopa, Patrick L. "Molecular mechanisms of nuclear factor-erythroid-2 related factor 2 (Nrf2) regulation phosphorylation by casein kinase 2 (CK2) and interaction with proto-oncogene N-Myc in neuroblastoma cells /." Morgantown, W. Va. : [West Virginia University Libraries], 2007. https://eidr.wvu.edu/etd/documentdata.eTD?documentid=5146.
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Thesis (Ph. D.)--West Virginia University, 2007.Title from document title page. Document formatted into pages; contains vi, 130 p. : ill. (some col.). Includes abstract. Includes bibliographical references.
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Assrir, Nadine. "Analyse des intéractions moléculaires entre deux protéines liant des histones, HIRA et HIRIP3 (HIRA-interacting protein 3) et leur partenaires respectifs, la protéine chaperon d'histones Asf1 et la sérine-thréonine kinase CK2." Paris 11, 2004. http://www.theses.fr/2004PA11T048.
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Wright, David C. "The role of PLC, cPKC, L-type calcium channels and CAMKII in insulin stimulated glucose transport in skeletal muscle." Virtual Press, 2002. http://liblink.bsu.edu/uhtbin/catkey/1233206.
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Julien, Mathéau A. "Mechanical Strain-Mediated Syndecan Regulation and Its Effects on Adhesion of Vascular Smooth Muscle Cells." Diss., Georgia Institute of Technology, 2005. http://hdl.handle.net/1853/7007.
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An injured vascular system has a substantial impact on an individuals overall health, and an understanding of the mechanisms that underlie blood vessel pathophysiology is required for the development of rational and effective treatment strategies. The phenotypic modulation of smooth muscle cells (SMC) during vascular injury, characterized by altered adhesion, migration and synthetic behavior, plays an important role in the eventual outcome. Specifically, the ability of SMCs to adhere to and remodel their extracellular environment via regulation of the syndecan class of cell adhesion molecules dictates the response of the vascular wall to local injury. The effect of in vitro syndecan-4 regulation on SMC adhesion was investigated through the use of a glass microsphere centrifugation assay, and an antisense-mediated reduction in gene expression was found to correlate with decreased adhesive strength. Regulation of syndecan-1, syndecan-2, and syndecan-4 gene expression was observed experimentally by mechanical strain of SMCs. Using real-time polymerase chain reaction (PCR), the kinetics of both static and cyclic mechanical strain were found to modify the gene expression in a time and strain magnitude-dependent manner unique to each syndecan. In particular, the responses of syndecan-4 were acute, but transient, while the evolution of syndecan-1 and syndecan-2 regulation was delayed by comparison. Mechanical strain also modulated syndecan-4 protein expression and ectodomain shedding, as measured by Western immunoblotting, and this effect was found, through selective inhibition, to be at least in part dependent on mitogen-activated protein (MAP) kinase signaling. In particular, intact extracellular signal-regulated MAP kinase (ERK) 1/2 and c-Jun NH2-terminal kinase / stress-activated protein kinase (JNK/SAPK) signaling pathways were found to be required for the observed strain-induced shedding. These findings offer a better understanding of syndecan function in response to mechanical strain and suggest potential new mechanisms by which physical forces may modulate vascular SMC behavior and regulation during normal physiology, pathologic conditions, and engineered arterial substitute development.
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Callaway, Kari-Kristin Anderson. "Mechanism and regulation of the protein kinase ERK2." Thesis, 2006. http://hdl.handle.net/2152/2470.
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Sayed, Mohamed Rabi. "Regulation and function of protein Kinase CK2 in cancer cells." Thesis, 2002. http://hdl.handle.net/2429/13322.
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Protein kinase CK2 is a ubiquitous eukaryotic protein serine/threonine protein
kinase present in both the nucleus and cytoplasm. In most cells, the holoenzyme form of
CK2 is a constitutively active heterotetramer of two catalytic (aand a') and two noncatalytic
P-subunits, but CK2(3-free pools of CK2a also exist. The activity of CK2 is
increased in rapidly growing tissues, tumor cells and human cancers. Despite the
identification of more than 160 substrates, many of which are important in cell division
and differentiation, the signaling pathways that regulate this enzyme have been
enigmatic. Here I provide evidence that protein kinase CK2 is regulated by the stressactivated
p38 MAP kinase signaling pathway. I found that p38 MAP kinase, in a
phosphorylation-dependent manner, can directly interact with and activate CK2
apparently through an allosteric mechanism.
The tumor suppressor protein p53 is one of the key molecules involved in
maintaining the stability of the genome, through both cell cycle arrest and apoptosis,
especially under circumstances of genotoxic stress. p53 has previously been shown to be
associated with protein kinase CK2, and undergo phosphorylation on Ser-392. However,
the physiological regulation and the functional consequences of this event remain
unknown. I found that this phosphorylation is dependent upon the stepwise activation of
p38 MAP kinase and CK2 in response to cellular stress. Furthermore, this activation was
demonstrated to be responsible for the biochemical regulation of the cyclin-dependent
kinase-1 (CDK-1), as both 5,6-dichloro-l-((3-D-ribofuranosyl)-benzimidazole (DRB; a
semi-specific inhibitor of CK2)) and antisense depletion of CK2, as well as SB203580 (a
specific inhibitor of p38 MAP kinase) were each associated with an inhibition of its
activation in response to nocodazole (microtubule inhibitor). I observed that depletion or
inhibition of the catalytic subunits of CK2, in the presence of microtubule inhibitor
(nocodazole), resulted in a compromise of the spindle assembly checkpoint.
Furthermore, the CK2-depleted cells exposed to nocodazole underwent reduced apoptosis
as well as abnormal duplication and alignment of centrosomes. These oncogenic
properties were observed in both human cervical carcinoma cells (i.e. HeLa cells) and
human colon tumor cells (HCT116). I also showed that this effect was dependent on the
presence of functional wild-type p53, as this phenomenon was not apparent in HCT116
p53"/_ cells. My results support a novel role for CK2 in concert with p53, to maintain the
stability of the genome. These findings may provide for an improved understanding of
abnormalities of the spindle checkpoint in human cancers, which is a prerequisite for
defining future therapies.
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Fahimdanesh, Kian. "Mining mutations for protein kinase CK2 across several cancer types." Thesis, 2019. https://hdl.handle.net/2144/36680.
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While the death toll of cancer has declined worldwide, the incidence is at an all-time high. Protein Casein kinase II (CKII) is a protein serine/threonine kinase that is essential to many cellular and biological functions in the mammalian organism. CK2 works as a heterotetramer protein consisting of two kinase and two regulatory subunits. Mutations in CK2 genes that lead to increased CK2 activity are associated with several human cancer types. To further our understanding of CK2’s relationship with cancer, we set to create and analyze a mutational profile for CK2 genes using a publicly available ATCG dataset via cBioPortal. For this, a collection of genomic data from patients with different cancer types was pooled from cBioPortal, allowing singular analysis of CK2 genes throughout different cancer types. Additionally, Kaplan-Meier plots assessed the effect of overall alterations in each CK2 gene and combinations of them. We found amino acid residue mutational hotspots in diverse cancer types. We also found some correlation between overall CK2 gene mutation and cancer patient disease-free survival and disease progression. Although our analysis is limited in its scope and specifics, however, it begs further investigation into the role of CK2 mutations in human malignancies.2025-06-14T00:00:00Z
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Shah, Anil Ajit. "Exploratory role of protein kinase CK2 synergy in treatment of breast cancer." Thesis, 2014. https://hdl.handle.net/2144/14684.
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Breast cancer is the second most common type of cancer among women. The serine-threonine protein kinase CK2 is overexpressed in many cancers, including lung, prostate, hematologic cancers, and breast (Pinna, 2013). Here, we examined the potential of CK2 inhibition alone and in combination with chemotherapy to treat breast cancer. We performed cell viability assays on five breast cancer cell lines treated with CK2 chemical inhibitors or small interfering RNAs, and chemotherapeutic drugs, to test if a synergistic effect could be attained. We also tested if CK2 inhibition would change the stem-like phenotype, epithelial-to-mesenchymal (EMT) marker expression, and CK2 subunit gene expression in the HS578T cell line. We concluded that in the five cell lines utilized, CK2 inhibition had no synergistic effect with chemotherapeutic drugs. CK2 inhibition had no effect on the stem-like phenotype of HS578T cells. However, CK2 inhibition did show a pattern of inhibition of EMT marker expression. Finally, we found that CK2 inhibition appears to activate a compensatory feedback loop for the transcription of the alpha subunit. This may explain the lack of synergy, and bears further investigation in future studies.
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Bosc, Denis G. "The catalytic subunits of protein kinase CK2, expression, covalent modification, and regulatory interactions." 1998. http://hdl.handle.net/1993/1578.
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Biochemical and genetic studies have demonstrated that protein kinase CK2 (CK2) is one component of the regulatory protein kinase network implicated in cell proliferation and cell cycle progression. Its ubiquitous distribution, lethal effects of gene disruption and high degree of conservation suggest an important role for CK2 in eukaryotic cells, although its exact role in cells remains poorly understood. CK2 is a protein serine/threonine kinase which is composed of two catalytic subunits (CK2_ and/or CK2_') and two regulatory subunits (CK2B). The CK2_ and CK2_' catalytic isozymes are encoded by different genes, are highly conserved between many species of animals and are structurally similar, but differ dramatically at their respective carboxyl terminal domains (CTDs). This observation, coupled with reports that CK2_ and CK2_' display different cell cycle dependent intracellular distributions and phosphorylation has led us to hypothesize that functional differences exist between CK2_ and CK2_'. In order totest this hypothesis we have: (i) examined the expression levels of these isozymes during cell cycle progression, (ii) used synthetic peptides and fusion proteins to identify the mitotic sites of phosphorylation within CK2_, and (iii) used the yeast two-hybrid system to identify potential regulatory protein partners. We have found that, although expression levels of CK2_ and CK2_ ' are relatively similar in cells progressing through the cell cycle, there is a dramatic increase in protein levels of both isozymes after stimulation of cells to enter the cell cycle. Moreover, there are increased levels of CK2B in mitotically arrested cells. These results suggest that egulation of the protein levels of the subunits of CK2 within cells is important for entry and exit of the cell cycle. We have identified Thr344, Thr360, Ser362, and Ser370 as the mitotic sites of phosphorylation on CK2_. Identification of these sites is an important step in defining the role of CKII and its phosphorylation during cell division. We have also identified a novel protein, Phu, which interacts specifically with CK2_, but not with CK2_' . The work presented in this thesis demonstrates that regulated protein levels of CK2_ and CK2_' are important for cell cycle entry, and that differences between CK2_ and CK2_ ' are reflected in isoform specific post-translational modifications and protein:protein interactions.
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張家銘. "The anti-apoptotic mechanisms of protein kinase CK2 in the brain of rat." Thesis, 2011. http://ndltd.ncl.edu.tw/handle/66053262671135584539.
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碩士國立政治大學
神經科學研究所
99
Protein kinase CK2 is a multifunctional serine/threonine protein kinase with many protein substrares and is ubiquitously expressed in mammalian cells to play an important role in cell cycle progression, transcription, and anti-apoptosis. In the nervous system, CK2 is shown to protect neurons against injury, but the cellular mechanisms are not well studies. In the present studies, we investigate which cellular mechanism might involve in the CK2 protection effects. The serum response factor (SRF) is a mammalian transcription factor which mediates some gene transcriptions relevent to promote the cell survival. The Myeloid cell leukemin 1 (Mcl-1) is one of the anti-apoptotic Bcl-2 family members and is involved in promoting cell viability. Previous studied have revealed that the SRF phosphorylation by CK2 can enhance its DNA-binding activity. The regulation of Mcl-1 by SRF has also been reported in other studies. In the first part of the present studies, we investigate whether the Mcl-1 expression is regulated by CK2 through SRF mediated pathway. The results from wildtype CK2α plasmid DNA transfection revealed that the phosphorylated SRF were increased in hippocampus CA1 region, whereas transfection of the catalytically inactive CK2αA156 mutant plasmid DNA decreased phosphorylated SRF. Further, wildtype CK2α increased, whereas CK2αA156 mutant decreased the mRNA and protein levels of Mcl-1. Moreover, transfection of the mutant SRF99A also decreased the mRNA and protein levels of Mcl-1. Furthermore, the mutant SRF99A antagonized the upregulatory effects of wildtype CK2α on Mcl-1 protein level in the co-transfection experiments. In the other side, DARPP-32 (dopamine- and cAMP-regulated phosphoprotein of Mr 32 kDa) is a signal transduction molecule that regulates the efficacy of dopamine signaling in neostriatal neurons. Previous studies have revealed that DARPP-32 might involve in the anti-apoptosis and its Ser102 residue is phosphorylated by CK2. Therefore, in the second part of this study, we investigate whether one of the anti-apoptotic effects of CK2 is through DARPP-32 phosphorylation by CK2 in the present study. The results revealed that the phosphorylated DARPP-32 is increased in stratum by wildtype CK2α transfection and decreased by catalytically inactive CK2αA156 mutant transfection. Further, transfection of CK2α siRNA can inhibit endogenous CK2 expression and also decrease phosphorylation of DARPP-32 as well as the anti-apoptotic protein, Bcl-xL. These results together suggest that CK2α-mediated anti-apoptotic effects are partially through SRF mediated or DARPP-32 mediated signaling to regulate Mcl-1 or Bcl-xL expression, respectively.
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Hamacher, Rainer W. [Verfasser]. "Die Protein-Kinase CK2 reguliert ein anti-apoptotisches Programm in Pankreaskarzinomzellen / Rainer W. Hamacher." 2008. http://d-nb.info/988564572/34.
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Dennis, Michael Don 1980. "Phosphorylation of plant translation initiation factors by CK2." Thesis, 2008. http://hdl.handle.net/2152/3868.
Full textAbstract:
Protein kinase CK2 phosphorylates wheat eIF2, eIF3, eIF4B, eIF5 and three 60S ribosomal proteins. The substrate specificity of CK2[alpha] toward various plant initiation factor substrates was altered in vitro through holoenzyme formation in the presence of regulatory [beta]-subunits. This presents a potential mechanism through which the differential expression and sub-cellular distribution of CK2 [beta]-subunits could regulate phosphorylation of various CK2 substrates in plants. Our analysis of initiation factor phosphopeptides produced by in vitro phosphorylation identified 20 CK2 phosphorylation sites in eIF2[alpha], eIF2[beta], eIF3c, eIF4B, and eIF5. Native wheat eIF5 was prepared in the presence of phosphatase inhibitors and analyzed by mass spectrometry. Native wheat eIF5 was determined to be a phosphoprotein containing at least 3 phosphorylation sites. The C-terminal CK2 site (S451) of native eIF5 was completely phosphorylated, and tryptic fragments containing the other in vitro CK2 two sites (S209, T240) also appear to be partially phosphorylated. Many of the CK2 phosphorylation sites identified are in conserved binding domains of the yeast multifactor complex (eIF1/eIF3/eIF5/eIF2/GTP/Met-tRNAi[superscript Met). This observation lead to the hypothesis that CK2 phosphorylation may regulate the formation of plant multifactor complexes. The results presented here suggest that plant initiation factors are capable of forming complexes similar to those previously reported in yeast. The in vitro interaction of initiation factors within these complexes appears to be enhanced by phosphorylation of eIF2, eIF3c, and eIF5 by CK2. Site-directed mutagenesis of eIF5 to remove CK2 phosphorylation sites not only prevents the CK2 mediated increase in interaction with eIF1, but also resulted in reduced stimulation of translation initiation in vitro.text
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Stark, Felix. "Funktionelle Untersuchungen zur Regulation der Protein Kinase CK2 durch Polyamine in Drosophila melanogaster und deren physiologische Bedeutung." Doctoral thesis, 2011. https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-57522.
Full textAbstract:
Die heterotetramere Proteinkinase CK2 nimmt aufgrund der großen Anzahl und Diversität ihrer Substrate, sowie aufgrund ihrer Eigenschaft Signalwege miteinander zu vernetzen eine Sonderstellung innerhalb der Kinasen ein. CK2 beeinflusst Proliferation, Differenzierung und Apoptose, Prozesse an denen auch Polyamine und der MAPK-Signalweg beteiligt sind. Eine vor kurzem durchgeführte Arbeit beschreibt die Bindung von CK2 an das Gerüstprotein KSR und die Verstärkung des MAPK-Signalwegs durch Phosphorylierung von Raf-Proteinen in Vertebraten. In dieser Arbeit konnte gezeigt werden, dass CK2 auch in Drosophila mit KSR interagiert und das einzige in Drosophila vorhandene Raf-Potein (DRaf) in vitro phosphoryliert. Im Gegensatz zur Phosphorylierung der humanen B-Raf und C-Raf Proteine an Serin 446 bzw. Serin 338 innerhalb der „negative charge regulatory region“ (N-Region), führten Kinasereaktionen und Massenspektrometrische Untersuchungen zur Identifizierung von Serin 11 als CK2 Phosphorylierungsstelle in DRaf, während ein zu Serin 446 in B-Raf äquivalentes Serin in der N-Region in Drosophila nicht durch CK2 phosphoryliert wird. Durch Überexpression von DRaf sowie von zwei DRaf-Varianten bei denen Serin 11 durch Alanin oder Aspartat substituiert wurde (DRafS11A und DRafS11D) konnte in Zellkulturexperimenten gezeigt werden, dass die Ladung an der Aminosäureposition 11 die Funktion von DRaf beeinflusst, wobei eine negative Ladung an dieser Stelle zur Phosphorylierung und Aktivierung der Effektorkinase Erk führt. Die Phosphorylierung durch CK2 ist unabhängig von regulatorischen Botenstoffen ("second messengers"), wird aber durch Bindung von Polyaminen moduliert. Intrazelluläre Polyamine entstammen zum grossen Teil dem zellulären Aminosäurekatabolismus und beeinflussen die Phosphorylierung von DRaf durch CK2 in vitro, wobei Spermin ein effizienter Inhibitor der Reaktion ist, während die Effekte von Putrescin und Spermidin gering sind. Auch in Drosophila Schneider S2 Zellen und in adulten weiblichen Fliegen hat Spermin einen inhibitorischen, CK2-abhängigen Effekt auf die Aktivierung von Erk. Ausserdem konnte gezeigt werden, dass Putrescin und Spermidin in der Lage sind die Aktivierung von Erk, im Vergleich zu Zellen die nur mit Spermin behandelt wurden, zu erhöhen. Das spricht dafür, dass die Phosphorylierung von DRaf und die davon abhängige Aktivierung von Erk durch CK2 von der Menge und Relation der verschiedenen Polyamine zueinander abhängt. Die Ergebnisse dieser Arbeit lassen den Schluss zu, dass der Polyaminmetabolismus über CK2 mit dem MAPK-Signalweg verknüpft ist. Nachdem Polyamine durch Aminosäurekatabolismus enstehen, kann auf diese Weise der MAPK-Signalweg in Abhängigkeit der Verfügbarkeit zellulärer Aminosäuren reguliert werden. Vorversuche zeigten eine Beeinflussung von Proliferation und Apoptose durch CK2 und Polyamine. Weitere Untersuchungen sind aber nötig um spezifische Einflüsse von Polyaminen und CK2 auf zelluläre Prozesse wie Proliferation, Differenzierung und Apoptose aufzudeckenBecause of its high number and diversity of substrates, as well as its ability to cross-link signalling pathways, the heterotetrameric protein kinase CK2 has an exceptional position within kinases. CK2 influences proliferation, differentiation and apoptosis, processes in which also polyamines and the MAPK-signalling pathway are involved. A recent publication delineates binding of CK2 to the scaffold protein KSR and the enhancement of the MAPK-signalling pathway by phosphorylation of Raf-proteins in vertebrates. In my thesis I could show that CK2 also interacts with KSR in Drosophila and phosphorylates the only existing Raf protein in Drosophila (DRaf) in vitro. In contrast to the phosphorylation of human B-Raf- and C-Raf-proteins on serine 446 respectively serine 338 within the "negative charge regulatory region" (N-region), kinase reactions and mass spectrometric analyses led to the identification of serine 11 as phosphorylation site in DRaf, whereas a serine in the N-region, which corresponds to serine 446 of B-Raf, is not phosphorylated by CK2 in Drosophila. In cell culture experiments overexpression of DRaf and two DRaf-variants, in which serine 11 was substituted by alanine or aspartate (DRafS11A and DRafS11D), revealed the charge at amino acid position 11 to affect the function of DRaf, with a negative charge leading to phosphorylation and activation of the effector kinase Erk. Phosphorylation by CK2 is independent of second messengers, whereas it is modified by binding of polyamines. Intracellular polyamines mainly derive from cellular amino acid catabolism and modulate the phosphorylation of DRaf by CK2 in vitro with spermine being an efficient inhibitor of the reaction, whereas the effects of putrescine and spermidine are minor. In Drosophila Schneider S2 cells and adult flies spermine inhibits the activation of Erk in a CK2-dependent way. Furthermore administration of putrescine and spermidine in combination with spermine leads to enhanced Erk activation in cells compared to cells that are treated with spermine. These results suggest that phosphorylation of DRaf and the subsequent activation of Erk by CK2 are dependent on the amount and relative concentrations of polyamines. Altogether the results of this work demonstrate a role for CK2 in linking polyamine metabolism to the MAPK-signalling pathway. Since polyamines derive from amino acid catabolism, the MAPK-signalling pathway can be regulated dependent on the availability of cellular amino acids. Preliminary experiments point to CK2- and polyamine-dependent effects on proliferation and apoptosis. Further investigations are necessary to reveal specific effects of polyamines and CK2 on cellular processes like proliferation, differentiation and apoptosis
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Lee, Hsiao Yi, and 李曉怡. "DARPP-32 phosphorylation by protein kinase CK2 mediates the anti-apoptotic effects in PC12 cells." Thesis, 2011. http://ndltd.ncl.edu.tw/handle/18623672044646569880.
Full textAbstract:
碩士國立政治大學
神經科學研究所
99
Protein kinase CK2 is a multifunctional serine/threonine protein kinase with many protein substrares and is ubiquitously expressed in mammalian cells. Many studies have shown that CK2 is involved in many neuronal functions including neuroprotection, but its cellular mechanisms are not well-studied. DARPP-32 (Dopamine- and cAMP-regulated phosphoprotein, Mr 32 kDa) is highly enriched in striatal medium-size spiny GABA neurons and is a prominent mediator of dopamine signalling which relates with drug abuse. Beside its well-known function in drug abuse, recent studies also reveal that DARPP-32 may be involved in the anti-apoptotic effects. Although the Ser102 residue of DARPP-32 is a phosphorylation site for CK2, this phosphorylation-mediated CK2 signaling has not been studied yet. The bcl-x gene, one member of the Bcl-2 family, encodes two isoform proteins Bcl-xL and Bcl-xS by the pre-mRNA alternative splicing. The former increases cell survival and the later enhances cell apoptosis. Our previous study found that CK2 can increase Bcl-xL expression by BDNF treatment. In the present study, we investigate whether DARPP-32 ser102 phosphorylation also mediates the CK2 signaling for cell survival. Our results revealed that DARPP-32 Ser102 phosphorylation, Bcl-xL protein level and Bcl-xL/Bcl-xS mRNA ratio were all increased by wild-type CK2α plasmid DNA transfection. Meanwhile, CK2 inhibitor TBB treatment or CK2α siRNA transfection decreased DARPP-32 Ser102 phosphorylation, Bcl-xL protein level and Bcl-xL/Bcl-xS mRNA ratio. On the other hand, DARPP-32 siRNA transfection decreased Bcl-xL protein level. Furthermore, transfection of DARPP-32 S102D, which mimics the constitutive phosphorylation form, increased whereas transfection of mutant S102A decreased the Bcl-xL protein level and Bcl-xL/Bcl-xS mRNA ratio. Further, the mutant DARPP-32 S102A antagonized the up-regulatory effects of wild-type CK2α on Bcl-xL protein level in the co-transfection experiments. From the results of H2O2-induced oxidative stress experiments, we also found that prior knock-down of CK2 or DARPP-32 can aggravate the decrease in DARPP-32 Ser102 phosphorylation, Bcl-xL protein level and Bcl-xL/Bcl-xS mRNA ratio by H2O2 treatment. These results together suggest that DARPP-32 mediates CK2α signaling in regulating Bcl-xL/Bcl-xS expression and this signaling pathway might be involved in cell survival under oxidative stress.
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Yang, Shu Ping, and 楊淑萍. "Neurotrophic factor BDNF up-regulates SRE-mediated gene transcription through protein kinase CK2 in PC12 cells." Thesis, 2010. http://ndltd.ncl.edu.tw/handle/37113819545025079395.
Full textAbstract:
碩士國立政治大學
生命科學研究所
98
The neurotrophins play an important role in cell differentiation and survival of the nervous system. Among them, the neuroprotective effects of brain-derived neurotrophic factor (BDNF) is showed to be mediated by extracellular signal-regulated kinase (ERK) and phosphatidylinositol-3 kinase (PI3K) signaling pathway in the recent studies. However, other cellular signaling pathways might be involved in these effects of BDNF. Protein kinase CK2 (casein kinase 2) is a ubiquitous and highly conserved serine/threonine protein kinase and is indicated as a vital cellular role. In recent years, evidences have been mounted in support of the importance of CK2 in the suppression of apoptosis. Serum response factor (SRF) is a transcription factor binding to a consensus DNA sequence SRE (known as a CArG box) which was found in the promoters of some immediately early genes (such as c-fos, Egr) and anti-apoptotic Mcl-1 gene. The activations of SRF-regulated genes were associated with cell proliferation, cell survival and perception of synaptic activity. However, the regulatory mechanism of SRE-mediated genes is not well studied. The SRE-mediated transcription activity through CK2 signaling by BDNF treatment was studied in the PC12 cells in the present study. Results revealed that BDNF significantly increased the SRE promoter activity by luciferase report assay. The SRE-mediated transcription activity was increased by overexpression of CK2α, and the inhibition of endogenous CK2α by small interfering RNA was also shown to reduce this transcription activity. Furthermore, CK2α siRNA treatment antagonized the up-regulation effects of BDNF on SRE-mediated transcription activity. The co-transfection of CK2 and mutant SRF S99A plasmids significantly diminished up-regulatory effects of CK2 on SRE promoter activity. To test this CK2 induction in SRE-mediated transcription plays a role in neuroprotecion, we determined whether over-expression CK2 protects PC12 cells against rotenone-induced apoptosis. The results revealed that the over-expression of CK2α protected cells against rotenone-induced apoptosis and rescued the SRE-mediated transcription activity. Further, these effects of CK2α were blocked by co-transfection of mutant SRF S99A. These above results demonstrate that the up-regulation of BDNF on SRE-mediated genes is through CK2 signaling pathway.