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Wong, Mei Wai Mie. "Functions of the golgin coiled-coil proteins of the Golgi apparatus". Thesis, University of Cambridge, 2014. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.708308.
Pełny tekst źródłaAu, Catherine. "Organellar proteomics of the Golgi apparatus and Golgi derived COPI vesicles". Thesis, McGill University, 2008. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=18742.
Pełny tekst źródłaLes techniques traditionnelles utilisées en biochimie, en histologie ainsi qu'en microscopie permettent la détermination d'un maximum de trois protéines à la fois dans l'étude d'une organelle. La mise en œuvre de la spectrométrie de masse en protéomique a complètement changé le panorama d'investigation des organelles. Pour la première fois, il est possible d'étudier le panel entier de protéines présent dans un compartiment sub-cellulaire. Dans cette étude, je démontre dans un premier temps que l'utilisation du dénombrement de peptides redondants permet la quantification des protéines et donc la capacité de comparer l'abondance relative de différentes protéines dans un échantillon complexe tel qu'une préparation d'organelle. Je présenterai par la suite le pipeline que nous utilisons pour isoler, caractériser et préparer les échantillons avant leur analyse par acquisition automatique par spectrométrie de masse laquelle est suivie par le traitement des données dont le résultat consiste à l'identification d'une liste de protéines. Un effort manuel important d'annotation est appliqué à cette liste préliminaire afin de générer un tableau final où sont assignées à la fois la fonction dans laquelle chaque protéine identifiée est impliquée, ainsi que l'attribution de la nomenclature la plus appropriée. Ce travail laborieux facilite par conséquent le dénombrement et l'attribution des peptides redondants aux protéines. Les organelles de la voie précoce de sécrétion sont analysées par le pipeline et après une vérification manuelle rigoureuse des données, les protéomes des microsomes rugueux, des microsomes lisses, de l'appareil de Golgi, et des vésicules dérivées du Golgi COPI GTP et COPI GTP?S sont déterminés. L'objectif de cette étude porte sur les protéomes du Golgi et des vésicules dérivées du Golgi dont les protéines les plus abondantes ainsi que leurs caractéristiques sont décrites en détail. L'hypoth
Dworkin, Joel. "Cell-free reconstitution of the Golgi apparatus". Thesis, McGill University, 1989. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=59884.
Pełny tekst źródłaHui, Hu. "Targeting and retention in the Golgi apparatus". Thesis, University College London (University of London), 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.263648.
Pełny tekst źródłaGilchrist, Annalyn. "Proteomics analysis of the endoplasmic reticulum and Golgi apparatus". Thesis, McGill University, 2008. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=18715.
Pełny tekst źródłaL'analyse par protéomique des microsomes du Réticulum Endoplasmique rugueux, des microsomes du Réticulum Endoplasmique lisse et de l'appareil de Golgi a permis l'identification de 1064 protéines par spectrométrie de masse. Par ailleurs, le fractionnement biochimique des microsomes lisses et rugueux par lavage avec une solution saline concentrée suivi d'un traitement au détergent Triton X-114 a permis l'identification de 598 nouvelles protéines. Les protéines furent quantifiées en fonction du nombre de peptides identifiés par spectrométrie de masse. La quantification des protéines a permis d'évaluer le degré de contamination croisée présent dans les fractions du Réticulum Endoplasmique, de l'appareil de Golgi et celui provenant des autres organelles. Les résultats de cette analyse ont révélé que la fraction de Golgi était contaminée jusqu'à un maximum de 20% par les protéines provenant du Réticulum Endoplasmique et que les mitochondries constituaient la source essentielle de contamination dans ces trois fractions. La clustérisation hiérarchique des protéines quantifiées a permis de dresser le profile de distribution des différentes protéines et ainsi de les assigner au sein des différents compartiments, à savoir aux microsomes du Réticulum Endoplasmique rugueux et/ou lisses, ou alors à l'appareil de Golgi. De ce fait, la protéine disulphide isomerase, ERp44, a été localisée dans l'appareil de Golgi. Ce résultat a été confirmé par immunolocalisation avec l'anticorps ERp44. Par ailleurs, cette même clustérisation hiérarchique a permis de localiser pour la première fois 176 protéines dans le Réticulum Endoplasmique correspondant ainsi à leur fonction putative prédite par bioinformatique. De plus, le fractionnement biochimique des microsomes lisses et rugueux a permis d'assigner les protéines dans les compartiments subcellulaires du Réticulum Endoplasmique : cytosol, membrane ou lumière. Ces résultats ont é
Rocchetti, Alessandra. "Interactions between the plant Golgi apparatus and the cytoskeleton". Thesis, Oxford Brookes University, 2016. https://radar.brookes.ac.uk/radar/items/e035b419-1acc-4031-aadd-2cfc1f9ed3c8/1/.
Pełny tekst źródłaShort, Benjamin. "Characterisation of rab-effector complexes at the Golgi apparatus". Thesis, University of Glasgow, 2003. http://theses.gla.ac.uk/31014/.
Pełny tekst źródłaRivinoja, A. (Antti). "Golgi pH and glycosylation". Doctoral thesis, University of Oulu, 2009. http://urn.fi/urn:isbn:9789514292699.
Pełny tekst źródłaAlbariri, Areej [Verfasser], i Thomas [Akademischer Betreuer] Kuner. "Golgi apparatus and Golgi outposts in neurons studied by correlative microscopy / Areej Albariri ; Betreuer: Thomas Kuner". Heidelberg : Universitätsbibliothek Heidelberg, 2020. http://d-nb.info/1220608793/34.
Pełny tekst źródłaLock, John George. "Dynamic imaging of post-Golgi protein transport /". [St. Lucia, Qld.], 2005. http://www.library.uq.edu.au/pdfserve.php?image=thesisabs/absthe19397.pdf.
Pełny tekst źródłaKam, Chuen. "Functional study of PICK1-ICA69 complex in the golgi apparatus /". View abstract or full-text, 2008. http://library.ust.hk/cgi/db/thesis.pl?NSNT%202008%20KAM.
Pełny tekst źródłaLian, Yen-Ling. "Mechanisms of retrograde transport from Golgi apparatus to endoplasmic reticulum". Electronic Thesis or Diss., Université Paris sciences et lettres, 2022. http://www.theses.fr/2022UPSLS067.
Pełny tekst źródłaMammalian cells are characterized by the co-existence of multiple pathways, including anterograde and retrograde transport. The Golgi apparatus has a central role in processing and sorting cargos in the bi-directional trafficking. The Retention Using Selective Hooks (RUSH) system allows to synchronize the transport of cargos from the ER to downstream compartments and to systematically analyze the secretory routes. Owing to the interaction of streptavidin (Str) and streptavidin-binding peptide (SBP), the reporter protein can be retained in the ER and then released by the addition of biotin. However, biotin has a high affinity to streptavidin, impairing reversibility of the RUSH assay. With the Artificial Ligands of Streptavidin (ALiS), Golgi-to-ER retrograde transport can be monitored using RUSH upon their washout.The reversible RUSH assay was firstly set up to study two mechanisms of Golgi-to-ER retrograde transport, including the KDEL-mediated retrieval pathway and the glycosylation enzyme recycling pathway. Core streptavidin fused with the ER-retention signal (Str-KDEL), or with the invariant chain (Ii-Str) were used as hooks. Golgi-resident enzymes, ManII* (Mannosidase II)-SBP-EGFP or ST* (Sialyltransferase)-SBP-GFP served as Golgi-targeted RUSH reporters. Our kinetic analysis showed that the Golgi-to-ER transport of ManII* and ST* are both slower through the glycosylation enzyme recycling pathway than the KDEL-meditated retrieval pathway. To characterize the role of putative regulatory factors in KDEL-mediated retrograde transport, we performed RNAi experiments targeting to COG3 and Rab6. Our data showed that knockdown of COG3 resulted in delayed retrograde transport of ManII* and ST*, and the depletion of Rab6 led to impaired trafficking of ST*, consistent with the previous studies. Our data indicated that there are two distinct pathways regulating the retrograde transport of Golgi cargos, including KDEL-mediated ER retrieval and ER recycling of Golgi glycosylation enzymes.Lastly, we have generated endogenous tagged Golgi glycosylation enzymes using CRISPR-Cas9 or CRISPaint knock-in approaches and applied the reversible RUSH assays in the selected clones. The subcellular distribution of endogenous ManIIEN-SBP-mNeonGreen, GalNAc-T1EN-SBP-mNeonGreen and B4GalT1EN-SBP-EGFP were detected in Golgi, and we were able to synchronize the bidirectional transport through the expression of Str-KDEL
Miranda, Kevin Charles. "Post-Golgi trafficking in the mammalian secretory pathway /". [St. Lucia, Qld], 2004. http://www.library.uq.edu.au/pdfserve.php?image=thesisabs/absthe18194.pdf.
Pełny tekst źródłaPecot, Matthew Y. "A declaration of independence the Golgi apparatus is here to stay /". Connect to a 24 p. preview or request complete full text in PDF format. Access restricted to UC campuses, 2006. http://wwwlib.umi.com/cr/ucsd/fullcit?p3225895.
Pełny tekst źródłaTitle from first page of PDF file (viewed October 8, 2006). Available via ProQuest Digital Dissertations. Vita. Includes bibliographical references (p. 107-115).
Poe, Tyler M., i Francine Marciano-Cabral. "Illumination of the Golgi apparatus of Pathogenic and Nonpathogenic Naegleria species". VCU Scholars Compass, 2019. https://scholarscompass.vcu.edu/etd/6002.
Pełny tekst źródłaNawazi, Fazlullah Salar Khan. "Golgi specificity and development of autoreactive B cells". View the abstract Download the full-text PDF version, 2008. http://etd.utmem.edu/ABSTRACTS/2008-019-Nawazi-index.html.
Pełny tekst źródłaTitle from title page screen (viewed on September 9, 2008). Research advisor: Marko Z. Radic, Ph.D. Document formatted into pages (xi,111 p. : ill.). Vita. Abstract. Includes bibliographical references (p. 91-111).
Zamzow, Daniel R. "Signaling capabilities of a novel H-Ras mutant from the Golgi apparatus". [Ames, Iowa : Iowa State University], 2006.
Znajdź pełny tekst źródłaWiggins, Christine Anne Ruth. "Identification and characterisation of proteins from the Golgi apparatus of S. cerevisiae". Thesis, University of Cambridge, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.624979.
Pełny tekst źródłaMarotta, D. "Defining the role of the Golgi apparatus in juvenile NCL (Batten disease)". Thesis, University College London (University of London), 2015. http://discovery.ucl.ac.uk/1460387/.
Pełny tekst źródłaHowe, Jonathon David. "Antiviral mechanisms of small molecules targeting the endoplasmic reticulum and Golgi apparatus". Thesis, University of Oxford, 2014. http://ora.ox.ac.uk/objects/uuid:04368b4b-2fd3-4fc7-8f89-ec39cd87e37d.
Pełny tekst źródłaKong, Anne Mandy 1973. "Cloning and characterisation of a novel 72 kDa inositol polyphosphate 5-phosphatase". Monash University, Dept. of Biochemistry and Molecular Biology, 2001. http://arrow.monash.edu.au/hdl/1959.1/9036.
Pełny tekst źródłaGuet, David. "Architecture of the Golgi apparatus and membrane trafficking probed by intracellular optical micromanipulation". Paris 6, 2012. http://www.theses.fr/2012PA066203.
Pełny tekst źródłaMembrane transport is based on the formation of tubulo-vesicular intermediates traveling from one compartment of the cell to another along cytoskeletal tracks. In vitro studies have shown that physical parameters, such as membrane curvature, tension and composition, influence the budding and fission of transport intermediates. Recent studies in cells have highlighted the central role of the actin cytoskeleton in the fission of transport intermediates from the Golgi apparatus. Here I investigate the role of a mechanical stress on intracellular transport in cellulo. I focus on the mechanics of Golgi membranes and the formation of transport intermediates from the Golgi apparatus. Using confocal microscopy, I visualize the deformation of Rab6-positive and COPI-positive Golgi membranes applied by an internalized microsphere trapped in an optical tweezers, and simultaneously measure the corresponding forces. My results show that the force necessary to deform Golgi membranes drops when the actin cytoskeleton is depolymerized, suggesting that actin strongly contributes to the local rigidity of the Golgi apparatus. We also show that the applied stress has a long-range effect on Golgi membranes and induces a sharp decrease in the formation of vesicles and the formation of tubular structures from the Golgi apparatus
Starr, Tregei Nicole. "Quantitative Studies of Intracellular Trafficking of Two Classes of Resident Golgi Apparatus Proteins". Diss., Virginia Tech, 2006. http://hdl.handle.net/10919/27217.
Pełny tekst źródłaPh. D.
Fuchs, Evelyn. "Regulation of Membrane Traffic at the Golgi Apparatus by Rab GTPases and their GAPs". Diss., lmu, 2008. http://nbn-resolving.de/urn:nbn:de:bvb:19-82832.
Pełny tekst źródłaAli, Zahabia Shameem. "Investigating the role of AMPK in carbohydrate sensing and localisation at the golgi apparatus". Thesis, Imperial College London, 2009. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.501761.
Pełny tekst źródłaFuchs, Evelyn. "Regulation of membrane traffic at the golgi apparatus by rab GTPases and their GAPs". kostenfrei, 2007. http://edoc.ub.uni-muenchen.de/8283/.
Pełny tekst źródłaDeng, Yuping. "Studies of intraorganelle dynamics : the lysosome, the pre-lysosomal compartment, and the golgi apparatus /". Diss., This resource online, 1991. http://scholar.lib.vt.edu/theses/available/etd-07282008-134815/.
Pełny tekst źródłaDahan, Sophie. "Intracellular proteinmembrane trafficking : evaluation of the Golgi and endosomal apparatus by cryoimmune electron microscopy". Thesis, McGill University, 1995. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=29387.
Pełny tekst źródłaWithin the secretory pathway, the hepatic Golgi apparatus was a site of protein concentration as evaluated by the gold labeling density of another major secretory protein of liver hepatocytes, albumin, which was concentrated $ sim$10-fold in the Golgi apparatus relative to the ER. Sorting of this secretory protein within pre-Golgi compartments was not observed. Within the Golgi apparatus, apoE was concentrated within Golgi saccular distensions while being predominantly absent from flattened saccular components; apoB was similarly segregated within peripheral distensions. In contrast albumin, as well as two other monomeric proteins, transferrin (Tf) and the polymeric immunoglobulin receptor (pIg-R) were distributed homogeneously throughout Golgi stacks. In an attempt to assess a key prediction of the vesicular transport hypothesis, small 60-90 nm vesicles in the immediate vicinity of Golgi apparatus, postulated to mediate intersaccular transport were examined for their content of cargo secretory or plasma membrane proteins. Lack of immunoreactive apoE, apoB, albumin, Tf, or pIgR, within small vasicular profiles suggests limits to current models of vesicle-mediated intra-Golgi transport.
Along the endocytic pathway, at the cell surface, apoE and pIgR were dispersely distributed along the sinusoidal microvilli. Quantitative analysis of the immunolabeling distribution of these proteins did not reveal concentration within plasma membrane pits. These findings which were confirmed by observations of cell surface labeling of two other ligands, Tf and apoB, are consistent with receptors and ligands gaining access to the endocytic machinery likely without receptor/ligand preclustering or prolonged clustering events within plasma membrane pits. Intracellularly, apoE was concentrated within endocytic structures which were double-labeled for apoE and internalized HRP. Large endocytic vesicles closely juxtaposed to Golgi stacks also revealed a high content of apoE. Together the endosomal labeling distribution of apoE as well as morphological features of endosomal components are consistent with the maturation model for endosomes.
Capitanio, Paola. "Effects of familial Alzheimer's disease-linked presenilin 2 mutants on Ca2+ homeostasis of Golgi Apparatus sub-compartments". Doctoral thesis, Università degli studi di Padova, 2013. http://hdl.handle.net/11577/3423438.
Pełny tekst źródłaLa malattia di Alzheimer's (AD) è un disordine neurodegenerativo e la forma più comune di demenza senile. La caratteristica istopatologica di AD è la presenza di depositi neurofibrillari intracellulari e di placche amiloidi, costituite da aggregati di peptide amiloide (Aß), che si depositano nella matrice extracellulare del cervello. I peptidi Aß sono il risultato di due tagli sequenziali della Proteina Precursore dell'Amiloide (APP); Aß viene poi rilasciato dall'enzima α-secretasi. Le più abbondanti specie peptidiche di Aß, prodotte anche fisiologicamente per tutta la vita, sono Aß40 e Aß42, quest'ultimo più insolubile e più incline all'aggregazione. Sebbene la maggior parte dei casi di AD siano sporadici, una piccola percentuale di pazienti è affetta dalla forma ereditaria di Alzheimer (malattia familiare di Alzheimer, FAD), causata da mutazioni dominanti in uno dei geni codificanti per APP, presenilina-1 (PS1) e presenilina-2 (PS2); le PSs sono le subunità catalitiche del complesso enzimatico della α-secretasi ma funzionano anche in maniera indipendente da tale attività enzimatica. Le mutazioni in PSs legate a FAD portano ad un aumento nel rapporto Aß42/Aß40, che promuove la deposizione di placche amiloidi. Oltre a questo effetto, è stato ampiamente dimostrato che molte mutazioni in PS1 e PS2 provocano alterazioni della omeostasi del Ca2+ intracellulare, rendendo così i neuroni più sensibili agli stimoli eccitotossici e apoptotici. L'apparato di Golgi (GA) rappresenta, insieme al reticolo endoplasmatico (ER), il principale deposito intracellulare di Ca2+, IP3 sensibile, e la sua funzionalità è fondamentale per il controllo delle risposte citosoliche di Ca2+. Sempre maggiori evidenze suggeriscono che il GA sia un organello eterogeneo in termini di Ca2+ handling, essendo dotato di un diverso toolkit molecolare per il Ca2+ rispetto a quello espresso nell' ER. Ad esempio, come meccanismi di uptake per il Ca2+, il GA esprime la classica pompa SERCA (Sarco-Endoplasmic Reticulum Ca2+ ATPase) ma anche un ulteriore pompa, detta SPCA1 (Secretory Pathway Ca2+ ATPase1). L'utilizzo di uno specifico sensore per il Ca2+ specificatamente indirizzato al trans-Golgi, ci ha precedentemente permesso di dimostrare direttamente la eterogeneità funzionale del GA, mostrando il comportamento distinto di questo sub-compartimento: i meccanismi di uptake di Ca2+ sono mediati esclusivamente dalla SPCA1 (e non dalla SERCA); non rilascia Ca2+ in risposta alla generazione IP3, ma piuttosto si accumula il catione come conseguenza dell'aumento di Ca2+ citoplasmatico. Per quanto riguarda gli altri sub-compartimenti del GA, abbiamo generato un nuovo indicatore per il Ca2+ fuso alla sequenza di indirizzamento dell'enzima 1,6 N-acetylglucosaminyltransferasi (C2gnT) residente del cis/medial-Golgi. La nuova sonda co-localizza con il marcatore di cis/medial-Golgi Giantina e quindi è stata utilizzata per studiare le dinamiche di Ca2+ in questo sub-compartimento a livello di singola cellula. Complessivamente i dati ottenuti suggeriscono che il GA sia unico in termini di omeostasi del Ca2+, con tali sub-compartimenti separati da pochi micron, e in equilibrio molto rapido tra loro, ma comunque in grado di mantenere differenze consistenti in termini di concentrazione dello ione e risposta a stimoli esterni . Le differenze tra i due sub-compartimenti del GA sono confermate dall'effetto specifico sulla omeostasi del Ca2+ dell'espressione della forma mutata di PS2T122R legata alla malattia familiare di Alzheimer. Le cellule che esprimono tale proteina mostrano una diminuzione del contenuto di Ca2+ nel cis/medial-Golgi ma nessun effetto sull'omeostasi del Ca2+ nel trans-Golgi. PS2T122R sembra inibire l'assorbimento di Ca2+ nel cis/medial-Golgi, inibendo l'attività della pompa SERCA, mentre non influenza l'assorbimento di Ca2+, mediato dalla SPCA1, nel trans-Golgi. Il GA sembra quindi giocare un ruolo importante nella patogenesi di AD e comprendere il contributo di tale organello nella patogenesi di AD e la sua base fisiopatologica potrà avere un forte impatto sulla possibilità di sviluppare terapie più efficaci per AD.
Fossati, M. "MECHANISMS OF PROTEIN TRANSPORT AT THE ER-GOLGI INTERFACE". Doctoral thesis, Università degli Studi di Milano, 2013. http://hdl.handle.net/2434/214981.
Pełny tekst źródłaFoote, Christopher. "The role of the AP-1 adaptor complex in trafficking between the trans-Golgi Network and endosomal system". Diss., Columbia, Mo. : University of Missouri-Columbia, 2005. http://hdl.handle.net/10355/4172.
Pełny tekst źródłaThe entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Title from title screen of research.pdf file viewed on (November 7, 2006) Vita. Includes bibliographical references.
Compton, Shannon Leigh. "Functional analysis of PRAF1 and its effect on corticotrophic ACTH secretion". Auburn, Ala., 2007. http://repo.lib.auburn.edu/07M%20Dissertations/COMPTON_SHANNON_44.pdf.
Pełny tekst źródłaWeigert, Roberto. "Biochemical analysis of the factors controlling the process of membrane tubule formation from the Golgi complex". Thesis, Open University, 2000. http://oro.open.ac.uk/58143/.
Pełny tekst źródłaYoung, Robin Elizabeth. "Secretion of plant cell wall polysaccharides by the Golgi apparatus in Arabidopsis thaliana seed coat cells". Thesis, University of British Columbia, 2009. http://hdl.handle.net/2429/11573.
Pełny tekst źródłaMillarte, Valentina [Verfasser]. "Signaling at the Golgi Apparatus During Cell Migration and Implication for Cancer Cell Metastasis / Valentina Millarte". Konstanz : Bibliothek der Universität Konstanz, 2015. http://d-nb.info/1080908919/34.
Pełny tekst źródłaHiding, Johan. "Functional characterization of the secretory pathway and the role of COPI vesicles /". Göteborg : Göteborg University, Institute of Biomedicine, Department of Medical Genetics, Sahlgrenska Academy, Göteborg University, 2007. http://hdl.handle.net/2077/8502.
Pełny tekst źródłaSzul, Tomasz J. "The role of GBF1 in Golgi biogenesis and secretory traffic". Thesis, Birmingham, Ala. : University of Alabama at Birmingham, 2009. https://www.mhsl.uab.edu/dt/2009p/szul.pdf.
Pełny tekst źródłaGrabski, Robert. "Using RNA interference to study the function of the tethering protein p115 in ER-Golgi traffic". Thesis, Birmingham, Ala. : University of Alabama at Birmingham, 2008. https://www.mhsl.uab.edu/dt/2008p/grabski.pdf.
Pełny tekst źródłaBellouze, Sarah. "Mécanismes moléculaires de la fragmentation de l' appareil de Golgi dans les maladies du neurone moteur". Thesis, Aix-Marseille, 2012. http://www.theses.fr/2012AIXM4080.
Pełny tekst źródłaFragmentation of the Golgi apparatus represents one of the earliest and most constant pathological changes in neurodegenerative diseases. To understand the molecular mechanisms of these changes I investigated two experimental models of motor neuron diseases. 1. pmn mice with progressive motor neuronopathy. The pmn mice were chosen since they suffer from a very aggressive form of motor neuron degeneration and since their molecular defects represents a missense mutation in a Golgi-localized tubulin chaperone TBCE, as shown by previous (Martin et al 2002, Schäfer et al 2007). In the last years, we identified severe Golgi abnormalities in motor neurons of pmn mice and dissected out their functional relevance and molecular mechanisms. According to immunolabelings and 3D membrane modelings, Golgi fragmentation and atrophy in lumbar pmn motor neurons resembled those reported in human ALS and proceeded with similar kinetics. Electron microscopy illustrated that Golgi cisternae were progressively transformed into small vesicles. Biochemical analyses revealed : 1/ a cytosolic redistribution of tethering factor such as GM130, 2/ a decrease in β-COP protein level and 3/ a massive increase in the Golgi v-SNARE proteins GS15 and GS28 controlling vesicle fusion. These pathological changes were due to loss of TBCE expression since they could be rescued by transgenic expression of wildtype TBCE but not mimicked by sciatic nerve axotomy. They involved defective dynamics of Golgi-derived microtubules rather than accumulation of misfolded tubulins as shown by the differential effects of TBCE-depletion, Nocodazole and a folding-incompetent tubulin mutant
Heymann, Julia. "Chlamydia infection impairs host cell motility via CPAF-mediated Golgi fragmentation". Doctoral thesis, Humboldt-Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, 2012. http://dx.doi.org/10.18452/16564.
Pełny tekst źródłaChlamydia are obligate intracellular human pathogens that proliferate inside a membrane-bound compartment called the inclusion. In infected cells, the Golgi apparatus is fragmented into small ministacks that are aligned around the inclusion. This facilitates uptake of host cell sphingolipids and is essential for chlamydial development. Infection-induced Golgi fragmentation happens after processing of the Golgi matrix protein golgin-84. This work could, via comparison with well-known substrates and inhibitor studies, identify the chlamydial protease CPAF (Chlamydia protease-like activity factor) as the enzyme accountable for this cleavage. Golgi Fragmentation depended on two Rab proteins, Rab6 and Rab11, which control vesicle transport and are recruited to the Chlamydia inclusion. As a consequence of Golgi fragmentation, cells lost the capacity to reorient the Golgi apparatus during polarization after a migratory stimulus. Both infected and golgin-84 depleted cells with a permanently fragmented Golgi apparatus displayed decelerated and furthermore randomized migration in a motility assay. Relocalization of the Golgi apparatus could be restored via stabilizing WEHD treatment or Rab depletion which partly rescued cell motility. Moreover, it could be shown that migration signaling via small GTPases was influenced by Chlamydia infection. Infected cells exhibited activation of the small polarity GTPase Cdc42. Numerous interactions with downstream effectors were strongly altered in infected cells according to quantitative mass spectrometry. Particularly, the binding of Cdc42 to migration-associated effectors was decreased. The results of this work show that CPAF, by processing of golgin-84, induces Golgi fragmentation which is vitally important for Chlamydia. This disturbs host cell motility because the Golgi apparatus cannot be reoriented during polarization and, additionally, via the modulation of protein-protein-interactions of Cdc42.
Bailly, Anne-Laure. "Rôle de GRASP-55 dans la spermatogenèse et la différenciation hématopoïétique". Thesis, Aix-Marseille, 2016. http://www.theses.fr/2016AIXM4112.
Pełny tekst źródłaThe junctional adhesion molecules JAM-B and JAM-C form a receptor / ligand pair involved in regulation of many biological mechanisms including inflammation, hematopoiesis and spermatogenesis. In the bone marrow, the interaction between JAM-C and JAM-B, expressed by hematopoietic stem cells (HSC) and stromal cells respectively, is involved in HSC retention and quiescence. Similarly, in the testis, JAM-C participates in the polarization of differentiated spermatids by interacting with JAM-B expressed by Sertoli cells. GRASP55 (Golgi ReAssembly and Stacking Protein of 55 kDa), identified in our laboratory as a new intracellular interactor of JAM, is a Golgi apparatus protein involved in Golgi architecture and dynamics as well as unconventional secretion.The aim of my thesis was to study the role of GRASP55 in vivo by genetic and pharmacological approaches. We demonstrate that GRASP55 expression by round spermatid allows polarized localization of JAM-C and the correct course of the spermatogenesis. In contrast, GRASP55 is not essential for hematopoiesis in basal or stress conditions. However, deletion of GRASP-55 in leukemic cells decreases the progression of the pathology in vivo. These results show a non-redundant role of GRASP55 in the spermatogenesis and proliferation of leukemic cells and allow us to consider GRASP55 as a potential target in hematology
Panić, Bojana. "The small GTPases Arl1p/Arl1 and Arl3p/ARFRP1 act in a pathway for targeting proteins to the Golgi apparatus". Thesis, University of Cambridge, 2004. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.616124.
Pełny tekst źródłaGreen, Harold G. "Ultrastructural and cytochemical characteristics of the Golgi apparatus in epithelial principal cells of the initial segment of the rat epididymis". Thesis, McGill University, 1990. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=59246.
Pełny tekst źródłaThe Golgi apparatus of these cells is composed of numerous saccular and tubular elements. On the cis face of the Golgi there is an osmiophilic tubular network. Of the numerous subjacent saccules, the first is dilated and unreactive for phosphatases, the next 3 show NADPase activity and the last 4 are TPPase positive. On the trans aspect of the stack there are 3 or 4 CMPase positive trans Golgi networks, one showing a peeling off configuration while the other 2 or 3 are completely separated from the stack. The TGN's often contain nodular distensions suggestive of secretion granule formation and occasionally present a shrivelled appearance.
Cisternae of ER showing buds are present on the cis face of the stacks.
Bruinsma, Paul. "The role of the yeast COG3, VPS35, and YDR141C proteins in membrane trafficking /". free to MU campus, to others for purchase, 2002. http://wwwlib.umi.com/cr/mo/fullcit?p3074381.
Pełny tekst źródłaMassarweh, Ahmad. "Dolichol linked Oligosaccharide Diphosphatase : a potential regulator of dolichol linked oligosaccharides". Thesis, Paris 6, 2016. http://www.theses.fr/2016PA066447.
Pełny tekst źródłaBACKGROUND: Type I congenital disorders of glycosylation (CDG-I) are caused by genetic defects in the biosynthetic pathway for the dolichol-linked oligosaccharide (DLO) that is required for protein N-glycosylation. These mutations result in the accumulation of truncated DLO and protein hypoglycosylation. Although protein hypoglycosylation is thought to be the main pathogenic factor in CDG-I, the role of truncated DLO intermediates in cellular homeostasis is not clear. Truncated DLO intermediates are known to give rise to cytoplasmic oligosaccharyl phosphates (OSP) by an uncharacterized mechanism. To understand this DLO editing process biochemical and molecular characterization of the activity that generate OSP is needed.RESULTS: I biochemically characterized a DLO diphosphatase (DLODP) that generates OSP and dolichol phosphate from DLO. Subcellular fractionation of mouse liver homogenates demonstrated a microsomal activity that co-distributes with a Golgi apparatus (GA) marker but not with endoplasmic reticulum (ER)-situated dolichol phosphate utilizing enzymes. This unexpected localization of DLODP prompted me to study OSP generation in cells using brefeldin A (BFA), which fuses the GA with the ER. BFA did not affect the levels of truncated DLO or cytoplasmic OSP, present in a cellular model of CDG-I. However, in these, and control cells, BFA caused striking increases of OSP within the endomembrane system. CONCLUSION: the identification of topologically distinct, independently modulated, OSP pools indicates multiple mechanisms for OSP generation and suggest that the GA-situated DLODP may not be the enzyme responsible for OSP generation in CDG-I
Serra, Peinado Carla. "Papel del citoesqueleto de actina en la regulación de la H+-ATPasa vacuolar de complejo de Golgi". Doctoral thesis, Universitat de Barcelona, 2014. http://hdl.handle.net/10803/663846.
Pełny tekst źródłaWe previously reported that agents that depolymerize actin filaments promote the alkalization of the Golgi stack and the trans-Golgi network. Vacuolar-type H-translocating ATPase (V-ATPase) is responsible of proton translocation and acidification of Golgi lumen. V-ATPase is a multisubunit complex composed of two domains (V1 and V0). Moreover, two subunits of V1 domain contain actin binding sides, subunit B and C. In this work we hypothesize that actin filaments could have a role in the maintaining of V1 and V0 domain association. We have generated a GFPtagged subunit B2 construct that is incorporated into the V1 domain, this construct localizes at distal Golgi compartments and translocate to cytosol upon actin depolymerization. Several biochemical assays confirmed that microfilaments distruption induces dissociation of V1-V0 domains. Moreover, we detected interaction between subunits B-C and actin filaments. Finally, V-ATPase is localized in lipid raft domains of plasma membrane and actin filaments participate in organization of these domains. We observed that lipid raft disorganization promotes an increase of intra-Golgi pH. Overall, we conclude that actin regulates the Golgi pH homeostasis maintaining the coupling of V1-V0 domains of V-ATPase through the binding of microfilaments to subunits B and C and preserving the integrity of lipid raft.
Ross, Kyla Turpin. "Quantitative Analysis of Feedback During Locomotion". Diss., Georgia Institute of Technology, 2006. http://hdl.handle.net/1853/14110.
Pełny tekst źródłaMarchini, Claudia Maria Meirelles. "O papel funcional da enzima fosfolipase D2 (PLD2) nas células da linhagem de mastócitos RBL-2H3". Universidade de São Paulo, 2008. http://www.teses.usp.br/teses/disponiveis/17/17136/tde-04122008-073156/.
Pełny tekst źródłaMast cells are components of the immune system that liberate a wide variety of pharmacologically active mediators. The principle method of activating mast cells is through the high affinity receptor for IgE (FcRI). This activation then culminates with the release of mediators. Phospholipase D (PLD) acts on phospholipids, hydrolyzing phosphatidylcholine to phosphatidic acid (PA) and choline. PLD is activated following stimulation via FcRI and plays an important role in signal transduction in mast cells. PLD has two isoforms, PLD1 and PLD2, which are differentially expressed depending on the cell type where none, one or both may be expressed. RBL-2H3 cells, a mast cell line, transfected to super express catalytically active (CA) and inactive (CI) forms of PLD2 were used in the present study. The role of PLD2 was examined in these cells in order to clarify the action of PLD2 in the secretory process. Although the CA and CI cells posses a greater total -hexosaminidase activity, when stimulated these cells release less -hexosaminidase than cells transfected with empty vector or wild type RBL-2H3 cells. In all cell lines, PLD2 was dispersed throughout the cytoplasm with a concentration in the juxtanuclear region suggesting an association of PLD2 with the Golgi apparatus. Double labeling with anti-PLD2 and mAb AA4, which recognizes gangliosides derived from GD1b on the plasma membrane, showed that PLD2 was not associated with the plasma membrane. When the cells were double labeled with anti-PLD2 and anti-GM130, which labels the cis-Golgi saccules, PLD2 does colocalize with the Golgi apparatus, especially in CI cells. Labeling with anti-GM130 alone as well as experiments employing transmission electron microscopy revealed that the Golgi apparatus is well organized in the CA cells, but is disorganized and dispersed in the cytoplasm in the CI cells. By Western Blotting, the CI cells also expressed less GM130 than the other cell lines. When the production of PA by PLD2 was inhibited by 1-Butanol, the Golgi apparatus of the CA cells presented the same phenotypic characteristics as that of the CI cells. Conversely, incubation of the CI cells with PA resulted in the reorganization of the Golgi apparatus. The structural maintenance of the Golgi apparatus is also related to microtubules. In the CI cells, the microtubule organizing center was difficult to identify and the microtubules were disorganized in the cytoplasm as compared to the other cell lines. These results show that the production of PA by PLD2 is important in the arrangement of the microtubules and in maintaining the structure of the Golgi apparatus. Alterations in the distribution of the microtubules and the structure of the Golgi apparatus in the CI cells affect the secretory process in these cells, and such alterations may affect the secretory process in other cell types as well. The findings presented here may lead to new therapeutic strategies to control the production and release of mediators during allergic and inflammatory processes.
Marais, Claire-Line. "Rôle de la SNARE Memb11 comme « récepteur » de la GTPase Arf1 à l’appareil de Golgi chez Arabidopsis thaliana". Thesis, Bordeaux 2, 2013. http://www.theses.fr/2013BOR22105/document.
Pełny tekst źródłaThe SNARE proteins (Soluble N-ethylmaleimide-sensitive factor Attachment protein REceptor) are critical for membrane fusion in the secretory pathway. I have studied the Golgi SNARE Memb11 in Arabidopsis thaliana cells. Memb11 is involved at the ER-Golgi interface. In mammalian cells, the ortholog of Memb11 (Membrin) is the potential “receptor” of the GTPase Arf1 in the Golgi membrane. This protein is involved for the recruitment of the COPI machinery, required for retrograde transport from the Golgi to the ER. The aim of this work was to determine whether Memb11 can interact with Arf1 in plant cells. Antibodies against the cytosolic part of Memb11 were obtained and were applied on plant tissues to perform immunolabeling by transmission electron microscopy and immunoprecipitation (IP) studies. It has been shown that Memb11 is located at the cis-Golgi and that it co-immunoprecipated with Arf1, suggesting that Arf1 may interact with Memb11. I confirmed the interaction of Memb11 and Arf1 at the Golgi by in vivo BiFC (Bimolecular Fluorescence Complementation) experiments. This interaction was specific since neither Memb12 (90% identity with Memb11) nor Sec22 interacted with ARF1. Thanks to a structural bioinformatic approach, I determined the regions in Memb11 (different from Memb12) that could be critical for the interaction and started to test corresponding mutants in vivo by BiFC. In addition, IP experiments with recombinant proteins produced in vitro suggest that the GTP-bound form of ARF1 interacts with Memb11
Despres, Julie. "Structure et fonctions de l'appareil de Golgi chez les fibroblastes dermiques humains lors du vieillissement : vers une stratégie innovante de criblaged'actifs dermo-cosmétiques à effets anti-age?" Thesis, Normandie, 2017. http://www.theses.fr/2017NORMR080.
Pełny tekst źródłaSkin is an important organ of the human body representing a protective structure in direct contactwith the external environment. During aging, skin undergoes dramatic changes including alteration ofdermal cells and components. Among these, fibroblasts synthetize and secrete a large variety ofcomponents and degrading enzymes involved in the modulation of dermal structure and functions. It isestablished that modification of the secreted components and enzymes during aging is related to dermisdegradation. This work aims to characterize aging-related alteration in fibroblasts. For this purpose, three aged human dermal primary fibroblast models have been developed. A transcriptomic study, using real-time quantitative PCR, has also been undertaken and has shown modifications in the expression of genesencoding dermal proteins. Using these results and in order to develop “anti-aging” cosmetic ingredients, extracts from polysaccharides-rich plant and microbial cells have been prepared and their efficiency evaluated on skin explants.As the Golgi apparatus is a major organelle of the secretory pathway, its structural organization has been investigated in fibroblasts using microscopy. The data show a marked alteration of trans-Golgi network morphology in aged cells. In contrast to its small and compact structure in young cells, the trans-Golgi network displays a large and expanded configuration in senescent cells. In addition, a transcriptomic analysis reveals that the expression of some genes, related to Golgi shape and/or function, is significantly modified in senescent cells. These genes could be then, used as innovating targets for the screening of novel dermo-cosmetic products with anti-aging activity