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Добірка наукової літератури з теми "Rac1 protein"

Автор: Grafiati
Опубліковано: 11 березня 2023

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Статті в журналах з теми "Rac1 protein"

1

Dumontier, M., P. Hocht, U. Mintert, and J. Faix. "Rac1 GTPases control filopodia formation, cell motility, endocytosis, cytokinesis and development in Dictyostelium." Journal of Cell Science 113, no. 12 (June 15, 2000): 2253–65. http://dx.doi.org/10.1242/jcs.113.12.2253.

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Анотація:
The function of the highly homologous Rac1A, Rac1B, and Rac1C GTPases of the Dictyostelium Rac1 group was investigated. All three GTPases bound with an equal capacity to the IQGAP-related protein DGAP1, with a preference for the activated GTP-bound form. Strong overexpression of wild-type Rac1 GTPases N-terminally tagged with green fluorescent protein (GFP), predominantly induced the formation of numerous long filopodia. Remarkably, expression of the constitutively-activated GTPases resulted in dominant-negative phenotypes: these Rac1-V12 mutants completely lacked filopodia but formed numerous crown shaped structures resembling macropinosomes. Moreover, these mutants were severely impaired in cell motility, colony growth, phagocytosis, pinocytosis, cytokinesis and development. Transformants expressing constitutively-inactivated Rac1-N17 proteins were similar to wild-type cells, but displayed abundant and short filopodia and exhibited a moderate defect in cytokinesis. Taken together, our results indicate that the three GTPases play an identical role in signaling pathways and are key regulators of cellular activities that depend on the re-organization of the actin cytoskeleton in Dictyostelium.
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2

Seiz, Julia R., Johannes Klinke, Laura Scharlibbe, Dirk Lohfink, Marisa Heipel, Hendrik Ungefroren, Klaudia Giehl, and Andre Menke. "Different signaling and functionality of Rac1 and Rac1b in the progression of lung adenocarcinoma." Biological Chemistry 401, no. 4 (March 26, 2020): 517–31. http://dx.doi.org/10.1515/hsz-2019-0329.

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AbstractRac1 is a ubiquitously expressed Rho GTPase and an important regulator of the actin cytoskeleton. Its splice variant Rac1b exhibits a 19-amino acid (aa) in-frame insertion and is predominantly active. Both proteins were described in tumorigenesis or metastasis. We investigated the contribution of Rac1 and Rac1b to tumor progression of human non-small-cell lung adenocarcinoma (NSCLA). Rac1 protein was present in 8/8 NSCLA cell lines analyzed, whereas Rac1b was expressed in only 6/8. In wound-healing assays, enhanced green fluorescence protein (EGFP)-Rac1 slightly decreased cell migration, whereas proliferation was increased in both, Rac1- and Rac1b-expressing cells. In the in vivo chorioallantoic invasion model, EGFP-Rac1-expressing cells formed more invasive tumors compared to EGFP-Rac1b. This increased invasiveness correlated with enhanced phosphorylation of p38α, AKT and glycogen synthase kinase 3β (GSK3β), and activation of serum response- and Smad-dependent gene promoters by Rac1. In contrast, Rac1b solely activated the mitogen-activated protein kinase (MAPK) JNK2, together with TCF/LEF1- and nuclear factor kappa B (NFκB)-responsive gene reporters. Rac1b, as Rac1, phosphorylated p38α, AKT and GSK3β. Knockdown of the splicing factor epithelial splicing regulatory protein 1 (ESRP1), which mediates out-splicing of exon 3b from Rac1 pre-messenger RNA, resulted in increased Rac1b messenger RNA (mRNA) and suppression of the epithelial-mesenchymal transition (EMT)-associated transcription factor ZEB1. Our data demonstrate different signaling and functional activities of Rac1 and Rac1b and an important role for Rac1 in lung cancer metastasis.
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3

Nagata, Koh-ichi, Yukio Okano, and Yoshinori Nozawa. "Differential Expression of Low Mr GTP-binding Proteins in Human Megakaryoblastic Leukemia Cell Line, MEG-01, and their Possible Involvement in the Differentiation Process." Thrombosis and Haemostasis 77, no. 02 (1997): 368–75. http://dx.doi.org/10.1055/s-0038-1655970.

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Анотація:
SummaryThe expression of various low Mr GTP-binding proteins at various states of differentiation of a human megakaryoblastic leukemia cell line, MEG-01, was analyzed using thermocycle amplification of mRNA and immunoblotting. MEG-01 cells were found to express mRNAs of rap1A, rap1B, rap2B, ralA, rhoA, rac1, rac2, CDC42Hs, rab1, rab3B, rab6, ram and ran, but not rab4, and the proteins of Rap 1, Rap2, RhoA, Rac1, Rac2, Rab3B, Rab4, Rab6 and Rab8 were expressed. Differentiation of MEG-01 cells induced by 100 nM 12-O-tetradecanoylphorbol-13-acetate revealed the considerable increases in mRNA expression of rap1B, rab3B, rabA, ram and ran whereas the levels of rap2B, rhoA and rac1 decreased. During the differentiation process, significant changes in protein levels of Rap1, RhoA, Rac1, Rac2, Rab3B, Rab4 and Rab6 were observed among three subcellular (cytosol, Triton X-100-soluble membrane and -insoluble cytoskeleton) fractions. The present investigation may be useful for the study of the megakaryocyte differentiation.
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4

Hoppe, Adam D., and Joel A. Swanson. "Cdc42, Rac1, and Rac2 Display Distinct Patterns of Activation during Phagocytosis." Molecular Biology of the Cell 15, no. 8 (August 2004): 3509–19. http://dx.doi.org/10.1091/mbc.e03-11-0847.

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Анотація:
The small G proteins Cdc42, Rac1, and Rac2 regulate the rearrangements of actin and membrane necessary for Fcγ receptor-mediated phagocytosis by macrophages. Activated, GTP-bound Cdc42, Rac1, and Rac2 bind to the p21-binding domain (PBD) of PAK1, and this interaction provided a basis for microscopic methods to localize activation of these G proteins inside cells. Fluorescence resonance energy transfer-based stoichiometry of fluorescent chimeras of actin, PBD, Cdc42, Rac1, and Rac2 was used to quantify G protein activation relative to actin movements during phagocytosis of IgG-opsonized erythrocytes. The activation dynamics of endogenous G proteins, localized using yellow fluorescent protein-labeled PBD, was restricted to phagocytic cups, with a prominent spike of activation over an actin-poor region at the base of the cup. Refinements of fluorescence resonance energy transfer stoichiometry allowed calculation of the fractions of activated GTPases in forming phagosomes. Cdc42 activation was restricted to the leading margin of the cell, whereas Rac1 was active throughout the phagocytic cup. During phagosome closure, activation of Rac1 and Rac2 increased uniformly and transiently in the actin-poor region of phagosomal membrane. These distinct roles for Cdc42, Rac1, and Rac2 in the component activities of phagocytosis indicate mechanisms by which their differential regulation coordinates rearrangements of actin and membranes.
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5

Kalfa, Theodosia A., Suvarnamala Pushkaran, Narla Mohandas, John H. Hartwig, Velia M. Fowler, James F. Johnson, Clinton H. Joiner, David A. Williams, and Yi Zheng. "Rac GTPases regulate the morphology and deformability of the erythrocyte cytoskeleton." Blood 108, no. 12 (December 1, 2006): 3637–45. http://dx.doi.org/10.1182/blood-2006-03-005942.

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Анотація:
Abstract Actin oligomers are a significant structural component of the erythrocyte cytoskeleton. Rac1 and Rac2 GTPases regulate actin structures and have multiple overlapping as well as distinct roles in hematopoietic cells; therefore, we studied their role in red blood cells (RBCs). Conditional gene targeting with a loxP-flanked Rac1 gene allowed Crerecombinase–induced deletion of Rac1 on a Rac2 null genetic background. The Rac1–/–;Rac2–/– mice developed microcytic anemia with a hemoglobin drop of about 20% and significant anisocytosis and poikilocytosis. Reticulocytes increased more than 2-fold. Rac1–/–;Rac2–/– RBCs stained with rhodamine-phalloidin demonstrated F-actin meshwork gaps and aggregates under confocal microscopy. Transmission electron microscopy of the cytoskeleton demonstrated junctional aggregates and pronounced irregularity of the hexagonal spectrin scaffold. Ektacytometry confirmed that these cytoskeletal changes in Rac1–/–;Rac2–/– erythrocytes were associated with significantly decreased cellular deformability. The composition of the cytoskeletal proteins was altered with an increased actin-to-spectrin ratio and increased phosphorylation (Ser724) of adducin, an F-actin capping protein. Actin and phosphorylated adducin of Rac1–/–;Rac2–/– erythrocytes were more easily extractable by Triton X-100, indicating weaker association to the cytoskeleton. Thus, deficiency of Rac1 and Rac2 GTPases in mice alters actin assembly in RBCs and causes microcytic anemia with reticulocytosis, implicating Rac GTPases as dynamic regulators of the erythrocyte cytoskeleton organization.
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6

Thomas, Emily K., Jose A. Cancelas, Heedon Chae, Adrienne D. Cox, Patricia J. Keller, Danilo Perrotti, Paolo Neviani, et al. "Rac GTPases Are Potential Therapeutic Targets in p210-BCR-ABL-Induced Myeloproliferative Disease (MPD)." Blood 110, no. 11 (November 16, 2007): 465. http://dx.doi.org/10.1182/blood.v110.11.465.465.

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Анотація:
Abstract The p210-BCR-ABL fusion protein is a constitutively active tyrosine kinase that is necessary and sufficient for the development of chronic myelogenous leukemia (CML). ABL-kinase inhibitors such as imatinib mesylate (Gleevec, STI571) potently block BCR-ABL activation, but the continued presence of leukemic stem cells and the emergence of imatinib-resistant BCR-ABL mutants suggest that ABL kinase inhibitors alone cannot completely eradicate disease. Rac GTPases have been implicated in BCR-ABL-mediated proliferation in cell lines and regulate many of the same signaling pathways as BCR-ABL, suggesting that these proteins could be additional therapeutic targets in CML. We have found that Rac1, Rac2, and, to a lesser extent, Rac3 were hyperactivated in CD34+ cells purified from the peripheral blood of two CML patients. To better study the role of Rac in BCR-ABL disease development, murine hematopoietic stem cells (HSC) genetically deficient in Rac1 and/or Rac2 were transduced with a retroviral vector expressing p210-BCR-ABL. Wild type (WT) and Rac1−/− mice experienced similar disease progression [median survival 23 ± 6 days (n=30) and 22 ± 4 days (n=8), respectively], Rac2−/− mice exhibited significantly attenuated development of BCR-ABL-mediated MPD [median survival 43 ± 27 days (n=18); p<0.001], and Rac1−/−;Rac2−/− animals showed markedly prolonged survival [median survival 92 ± 34 days (n=19); p<0.001]. p210-BCR-ABL WT, Rac1−/−, and Rac2−/− mice had elevated circulating myeloblasts 30 days post-transplant, while Rac1−/−;Rac2−/− mice had normal peripheral blood morphology. Attenuation of disease in Rac2- and Rac1/Rac2-deficient animals correlated with severely diminished activation of BCR-ABL-induced signaling pathways, including p44/42 and p38 ERK, JNK, CrkL, and Akt. The leukemogenesis impairment induced by Rac deficiency did not appear to be due to loss of p210-BCR-ABL vector integration, as clonal analysis of leukemic bone marrow from mice in each genotype by LAM-PCR showed similar, oligoclonal reconstitution of p210-BCR-ABL expressing cells. Interestingly, bone marrow cells obtained from Rac1/Rac2-deficient animals that developed late leukemia showed marked hyperactivation of Rac3 and initiated disease in recipients with a latency of three weeks, suggesting that leukemia-initiating cells were able to engraft, in spite of Rac1/Rac2 deficiency. Treatment of BCR-ABL-expressing murine HSC with NSC23766, a rationally-designed Rac-specific small molecule antagonist, potently inhibited cell proliferation in vitro and increased the survival of leukemic animals treated in vivo, compared to PBS control-treated animals (p<0.05). NSC23766 also inhibited the growth of an imatinib-resistant p210-BCR-ABL-T315I-expressing Ba/F3 leukemic cell line by 90%, compared to <5% by imatinib alone, blocked the growth of primary human chronic phase Rac-hyperactivated CML blast colonies by 80% in vitro, and inhibited survival of these cells in NOD-SCID mice. These results suggest that individual Rac proteins play both unique and combinatorial roles in stem cell transformation and may represent unique targets for therapy of BCR-ABL-persistent and imatinib-resistant CML.
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7

Chuang, T. H., X. Xu, L. A. Quilliam, and G. M. Bokoch. "SmgGDS stabilizes nucleotide-bound and -free forms of the Rac1 GTP-binding protein and stimulates GTP/GDP exchange through a substituted enzyme mechanism." Biochemical Journal 303, no. 3 (November 1, 1994): 761–67. http://dx.doi.org/10.1042/bj3030761.

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Анотація:
The Rac proteins, Rac1 and Rac2, are essential components of the NADPH oxidase system of phagocytes and regulate the actin assembly associated with membrane ruffling. These functions are controlled by the GTP-bound form of Rac. The biochemical interaction between Rac and its only known GDP-dissociation stimulator (termed smgGDS) was characterized. SmgGDS was able to stimulate the incorporation of guanosine 5′-[gamma-thio]-triphosphate GTP[gamma S] into the RhoA, Rac2, Rac1, Rap1A and CDC42Hs GTP-binding proteins, but the activity was greatest toward RhoA and Rac2. Isoprenoid modification of these proteins was not absolutely required for the interaction with smgGDS. Interestingly, the activity of smgGDS toward Rac1 could not be observed in a [3H]GDP/GTP exchange assay under conditions where it stimulated incorporation of GTP[gamma S] into Rac1. We determined that smgGDS prevented the loss of Rac1 activity during the [3H]GDP/GTP exchange assay by demonstrating the ability of smgGDS to inhibit the loss of Rac1 GTP[gamma S]-binding during incubations at 30 degrees C. This stabilizing effect was exactly counterbalanced by the ability of smgGDS to stimulate the release of [3H]GDP from Rac1, thereby producing no net observable effect in the exchange assay. SmgGDS was able to effectively stimulate the release of GDP but not GTP[gamma S] from Rac1. SmgGDS maintains Rac1 in a nucleotide-free form after release of GDP, indicating that the reaction between Rac1 and smgGDS involves a substituted enzyme mechanism.
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8

Engers, R., S. Ziegler, M. Mueller, A. Walter, R. Willers, and H. E. Gabbert. "Prognostic relevance of increased Rac GTPase expression in prostate carcinomas." Endocrine-Related Cancer 14, no. 2 (June 2007): 245–56. http://dx.doi.org/10.1677/erc-06-0036.

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Анотація:
Rac proteins of the Rho-like GTPase family, including the ubiquitous Rac1, the hematopoiesis-specific Rac2, and the least-characterized Rac3 play a major role in oncogenic transformation, tumor invasion and metastasis. However, the prognostic relevance of Rac expression in human tumors has not been investigated yet. In the present study, Rac protein expression was analyzed in benign secretory epithelium, high-grade prostatic intraepithelium neoplasia (HG-PIN), and prostate carcinomas of 60 R0-resected radical prostatectomy specimens by semiquantitative immunohistochemistry. Thus, Rac proteins were significantly strongly expressed in HG-PIN (P < 0.001) and prostate carcinomas (P < 0.001) when compared with benign secretory epithelium. Accordingly, all tumor tissues analyzed by isoform-specific real-time PCR (n = 7) exhibited significantly higher RNA expression levels of Rac (i.e. sum of Rac1 and Rac3 expression levels) than the respective benign counterparts (P = 0.018) and this appeared to result mainly from increased expression of the Rac3 isoform as verified by immunoblotting. Univariate analyses showed statistically significant associations of increased Rac protein expression in prostate cancer (P = 0.045), preoperative prostate-specific antigen levels (P = 0.044), pT stage (P = 0.002), and Gleason score (P = 0.001) with decreased disease-free survival (DFS). This prognostic effect of increased protein expression of Rac remained significant even in a multivariate analysis including all these four factors (relative risk = 3.22, 95% confidence interval = 1.04–10.00; P = 0.043). In conclusion, our data suggest that increased Rac protein expression in prostate cancer relative to the corresponding benign secretory epithelium is an independent predictor of decreased DFS and appears to result mainly from increased expression of the Rac3 isoform.
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9

Kuncewicz, Teresa, Priya Balakrishnan, Mark B. Snuggs, and Bruce C. Kone. "Specific association of nitric oxide synthase-2 with Rac isoforms in activated murine macrophages." American Journal of Physiology-Renal Physiology 281, no. 2 (August 1, 2001): F326—F336. http://dx.doi.org/10.1152/ajprenal.2001.281.2.f326.

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Анотація:
Nitric oxide synthase-2 (NOS2) is responsible for high-output nitric oxide production important in renal inflammation and injury. Using a yeast two-hybrid assay, we identified Rac2, a Rho GTPase member, as a NOS2-interacting protein. NOS2 and Rac2 proteins coimmunoprecipitated from activated RAW 264.7 macrophages. The two proteins colocalized in an intracellular compartment of these cells. Glutathione- S-transferase (GST) pull-down assays revealed that both Rac1 and Rac2 associated with GST-NOS2 and that the NOS2 oxygenase domain was necessary and sufficient for the interaction. [35S]methionine-labeled NOS2 interacted directly with GST-Rac2 in the absence of GTP, calmodulin, or NOS2 substrates or cofactors. Stable overexpression of Rac2 in RAW 264.7 cells augmented LPS-induced nitrite generation (∼60%) and NOS2 activity (∼45%) without measurably affecting NOS2 protein abundance and led to a redistribution of NOS2 to a high-speed Triton X-100-insoluble fraction. We conclude that Rac1 and Rac2 physically interact with NOS2 in activated macrophages and that the interaction with Rac2 correlates with a posttranslational stimulation of NOS2 activity and likely its spatial redistribution within the cell.
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10

Michaelson, David, Joseph Silletti, Gretchen Murphy, Peter D'Eustachio, Mark Rush, and Mark R. Philips. "Differential Localization of Rho Gtpases in Live Cells." Journal of Cell Biology 152, no. 1 (January 8, 2001): 111–26. http://dx.doi.org/10.1083/jcb.152.1.111.

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Анотація:
Determinants of membrane targeting of Rho proteins were investigated in live cells with green fluorescent fusion proteins expressed with or without Rho-guanine nucleotide dissociation inhibitor (GDI)α. The hypervariable region determined to which membrane compartment each protein was targeted. Targeting was regulated by binding to RhoGDIα in the case of RhoA, Rac1, Rac2, and Cdc42hs but not RhoB or TC10. Although RhoB localized to the plasma membrane (PM), Golgi, and motile peri-Golgi vesicles, TC10 localized to PMs and endosomes. Inhibition of palmitoylation mislocalized H-Ras, RhoB, and TC10 to the endoplasmic reticulum. Although overexpressed Cdc42hs and Rac2 were observed predominantly on endomembrane, Rac1 was predominantly at the PM. RhoA was cytosolic even when expressed at levels in vast excess of RhoGDIα. Oncogenic Dbl stimulated translocation of green fluorescent protein (GFP)-Rac1, GFP-Cdc42hs, and GFP-RhoA to lamellipodia. RhoGDI binding to GFP-Cdc42hs was not affected by substituting farnesylation for geranylgeranylation. A palmitoylation site inserted into RhoA blocked RhoGDIα binding. Mutations that render RhoA, Cdc42hs, or Rac1, either constitutively active or dominant negative abrogated binding to RhoGDIα and redirected expression to both PMs and internal membranes. Thus, despite the common essential feature of the CAAX (prenylation, AAX tripeptide proteolysis, and carboxyl methylation) motif, the subcellular localizations of Rho GTPases, like their functions, are diverse and dynamic.
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Більше джерел

Дисертації з теми "Rac1 protein"

1

Schnelzer, Andreas Christof. "Untersuchungen zum kleinen G-Protein Rac1 beim Mammakarzinom." [S.l.] : [s.n.], 2001. http://deposit.ddb.de/cgi-bin/dokserv?idn=963997610.

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2

Wang, PiLin. "Roles for Small G Protein Rac1 in Amyloid Cascade Hypothesis." 京都大学 (Kyoto University), 2009. http://hdl.handle.net/2433/124063.

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3

Hope, Hannah. "Regulation of the Rho G protein, Rac1, in the pathogenic fungus Candida albicans." Nice, 2009. http://www.theses.fr/2009NICE4093.

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Анотація:
C. Albicans est un pathogène opportuniste de l’homme, en général présent dans la flore intestinale. Un des facteurs de virulence de cet organisme est sa capacité de dimorphisme: cette levure évolue entre une forme ovoïde et une forme filamenteuse invasive. Ce changement de forme requiert une réorganisation du cytosquelette, suggérant que les facteurs clés de cette réorganisation - les protéines G de type Rho et leurs régulateurs - jouent un rôle important dans la croissance filamenteuse et la virulence de cet organisme. A la différence de l’homme, il n’y a que six de ces protéines G chez la levure. L’objectif de ce travail est d’étudier le rôle et la régulation de l’une d’entre elles, Rac1, identifiée précédemment dans le laboratoire. Nous avons tout d’abord identifié l’activateur de Rac1, Dck1, et nos résultats suggèrent que la spécificité de Rac1 est en partie déterminée par cet activateur. Dck1 fait partie de la famille d’activateurs CDM, définie par C. Elegans Ced-5, H. Sapiens Dock180 et D. Melanogaster Myoblast city. Chez les mammifères, la protéine d’échaffaudage ELMO (engulfment and cell motility) est nécessaire, en plus de Dock180, pour l’activation optimale de Rac1. Une recherche BLAST du génome de C. Albicans a permis d’identifier un analogue d’ELMO1, appelé LMO1. L’étude de la régulation de Rac1 a été poursuivie en caractérisant le rôle de Lmo1, utilisant des approches génétiques et biochimiques. Nos résultats indiquent qu’un mutant de délétion lmo1 est déficient pour la croissance filamenteuse invasive, tout comme les mutants de délétion rac1 et dck1. La sur-expression spécifique de la protéine MAP Kinase Cek1 permet de restaurer la croissance filamenteuse invasive dans chacun de ces mutants de délétion. De plus, nos résultats montrent que Rac1, Dck1 et Lmo1 sont critiques pour l’intégrité cellulaire. D’autre part, nous avons pu déterminer que Lmo1 lie Dck1 et Rac1, soit ensemble ou soit separament. . Dans l’ensemble, nos résultats permettent de proposer que Rac1, Dck1 et Lmo1 fonctionnent en complexe, en amont de la MAP kinase Cek1, lors de la croissance filamenteuse invasive de C. Albicans. Cette étude montre pour la première fois la conservation d’un complexe fonctionnel Dock180/ELMO/Rac1 de la levure à l’homme
C. Albicans is a human opportunistic pathogen that can invade host tissues, causing infections ranging from superficial epithelial mycoses to life threatening systemic infections in immuno-compromised patients. C. Albicans is a dimorphic fungus that exists either as an oval budding yeast or as a filamentous form (hyphal or pseudohyphal). The morphological switch between these two states is critical for C. Albicans pathogenicity. Rho G-proteins are well-documented regulators of the actin cytoskeleton and cell polarity in all eukaryotic cells. In C. Albicans it has been previously shown that distinct Rho G-proteins are required for filamentous growth in response to specific stimuli. Cdc42 is essential for filamentous growth in response to serum whereas Rac1 is required for filamentous growth in an agar matrix. The aim of my thesis research was to determine how Rac1 is regulated during filamentous growth in C. Albicans. First, we identified Dck1, which belongs to the Ced-5, Dock180, myoblast city (CDM) family of guanine nucleotide exchange factors, which is required for invasive filamentous growth similar to Rac1. Subsequently we showed using in vitro binding and genetic suppression studies that Dck1 can activate Rac1 during invasive filamentous growth. From these results we concluded that the requirement for distinct G-proteins for filamentous growth in reponse to different stimuli is due to the specificity of their activators. In mammals, flies and worms the ELMO (engulfment and cell motility) protein functions as a scaffold which is necessary for Rac1 signalling. Based on similarity to the mammalian ELMO1 we identified Lmo1 and showed that it is required for invasive filamentous growth. Furthermore we demonstrated that Lmo1 interacts with both Dck1 and Rac1. Overexpression of the MAP kinase Cek1 restores invasive filamentous growth in each of these three deletion mutants. In addition we identified a role for these proteins in cell integrity. Together, our results indicate that Lmo1, Rac1 and Dck1 function as a complex upstream of the Cek1 MAP kinase during invasive filamentous growth. This work shows for the first time the conservation of the Dock180/ELMO/Rac1 signalling complex between fungi and mammals
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4

Schenck, Annette. "CYFIP, a protein family implicated in neuronal connectivity, links Rac1 GTPase signalling to the fragile X mental retardation protein." Université Louis Pasteur (Strasbourg) (1971-2008), 2003. http://www.theses.fr/2003STR13175.

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Анотація:
Le syndrome de l'X Fragile, qui constitue la forme la plus fréquente de retard mental héréditaire, est du à l'absence de FMRP, une protéine de liaison à l'ARN qui régulerait la traduction au niveau synaptique. Afin de mieux comprendre le role de FMRP, nous avons réalisé un criblage double-hybride dans la levure pour identifier certains de ses interacteurs. Deux protéines ont été isolées, CYFIP1 et CYFIP2 (Cytoplasmic FMRP Interacting Proteins 1/2). Ces deux protéines cytoplasmiques sont hautement homologues mais interagissent différement avec les deux autres protéines de la famille FXR, FXR1P et FXR2P. Le site de liaison à CYFIP recouvre le site d'homo- et d'hétérodimérisation de FMRP, suggérant que la liaison à CYFIP pourrait moduler l'activité de FMRP. D'autre part, l'interaction entre CYFIP1 et Rac1 a été démontrée précédemment. Rac1, une protéine de la famille des Rho GTPase, est un des principaux régulateurs de la réorganisation du cytosquelette d'actine et joue un role clé dans la maturation et la maintenance des épines dendritiques, structures synaptiques riches en actine anormalement développées chez les patients X-Fragile et dans les souris invalidées pour FMRP. De nombreux gènes des voies Rho/Rac étant impliqués dans des retards mentaux, Rac1, CYFIP1 et FMRP pourraient participer à une voie commune contrôlant la morphogénèse synaptique et le fonctionnement cognitif. Pour valider cette hypothèse in vivo, nous avons choisi Drosophila melanogaster comme organisme modèle. Nous avons découvert que CYFIP y est très exprimé dans le système nerveux embryonnaire, et s'accumule notamment dans les axones centraux et aux jonctions neuro-musculaires (JNM). Des mutations de CYFIP causent des défauts de croissance, de ramification et de connexion des axones et conduisent à une morphologie anormale des synapses des JNM. Ainsi, l'absence de CYFIP provoque des défauts similaires à ceux précedemment décrits chez les mutants dFMR1 et/ou dRac1. L'analyse des interactions génétiques et biochimiques entre ces trois protéines suggère que, lorsque la voie est activée, dRac1 inhibe CYFIP qui régule à son tour négativement dFMR1
Fragile X Syndrome is the most frequent form of hereditary mental retardation and caused by the absence of FMRP, an RNA binding protein that seems to regulate local protein translation at synapses. To better understand the physiological function of FMRP, we conducted a yeast two-hybrid screen to determine interacting proteins. We identified CYFIP1 and CYFIP2 (Cytoplasmic FMRP Interacting Proteins 1/2), two highly homologous cytoplasmic proteins, which show a different pattern of interaction with the two FMRP-related proteins FXR1P and FXR2P. The CYFIP binding site of FMRP overlaps with its homo- and heteromerisation domain, suggesting that binding to CYFIP may modulate FMRP function. Importantly, CYFIP1 has been previously reported to interact with Rac1. Rac1, a Rho GTPase, is a key regulator of actin cytoskeleton remodelling with a well-established role in maturation and maintenance of dendritic spines, which are actin-rich synaptic structures that are abnormally developed in Fragile X patients and FMRP null mice. Since several genes of Rac/Rho signalling pathways are implicated in mental retardation, our work suggested that Rac1, CYFIP and FMRP work in a common pathway determining synapse morphogenesis and cognitive function. To address this hypothesis in vivo, we have chosen the fruitfly Drosophila melanogaster as a genetic model organism. Drosophila CYFIP, a previously undescribed gene, is highly expressed in the embryonic nervous system, where it strongly accumulates in central axons and at the neuromuscular junction (NMJ). CYFIP mutations induce defects in axon growth, branching and pathfinding and result in abnormal synapse morphology at the neuromuscular junction. Hence, loss of CYFIP involves defects that have been previously described in dFMR1 and/or dRac1 mutants. Analyses of biochemical and genetic interactions amongst these three proteins suggest that upon activation, dRac1 acts antagonistically on CYFIP, which in turn negatively regulates dFMR1
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Mehta, Harshini. "Role of adaptor protein SLAT in Fc[gamma]R mediated phagocytosis in macrophages." Oklahoma City : [s.n.], 2009.

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Attias, Ortal. "The role of Rac1 in mouse podocyte cellular process formation and differentiation /." Thesis, McGill University, 2008. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=111588.

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The role of podocytes in glomerular permselectivity is tightly associated with their intricate morphology, featuring interdigitating foot processes from adjacent cells. The actin cytoskeleton is an integral component of podocyte foot processes and is regulated by a number of proteins expressed in podocytes. Rho-family of small GTPases are known key regulators of the actin cytoskeleton. This study, investigated the role of Rac1 in podocytes, using conditionally immortalized mouse podocytes (MPs). We studied Rho-GTPase activities and morphology/cytoskeleton of differentiating mouse podocytes stably expressing nephrin. We also studied the impact of transfection of various Rho-GTPase mutants and IQGAP1 mutants. We demonstrated that nephrin expression potentiates and sustains Rac1 activity during the differentiation process. We showed that Rac1 contributes to process formation in differentiating MPs and may have a similar role in vivo when podocytes are recovering from injuries.
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Saito, Kelly Cristina. "Envolvimento de Rac1 na excitotoxicidade induzida por NMDA na retina de ratos." Universidade de São Paulo, 2011. http://www.teses.usp.br/teses/disponiveis/42/42134/tde-10022012-132908/.

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A ativação excessiva dos receptores NMDA tem sido descrita no disparo da morte neuronal que ocorre em doenças, como o glaucoma. É possível que a combinação de subunidades (NR2A-D) possa ativar vias de sinalização intracelulares que resultam na morte ou sobrevivência. Nosso objetivo foi investigar o envolvimento de subunidades NR2 e Rac1, membro da família Rho GTPase, na morte de neurônios da retina. A morte induzida por glutamato in vitro foi reduzida após a inibição de Rac1 e bloqueio de NR2B, mas não das subunidades NR2C/D. Resultados semelhantes foram obtidos in vivo após injeção intravítrea NMDA, e a detecção de Rac1 ativo, principalmente, nos processos de glia de Müller foi inibida pelo bloqueio NR2B. Além disso, a produção de TNF-α após a injeção de NMDA foi reduzida pelo bloqueio de NR2B e Rac1. Assim, nossos resultados sugerem que a excitotoxicidade via receptores NR2B/NMDA ativa Rac1 em células da glia de Müller, que por sua vez controla a produção de TNF-α possível responsável pela morte de células ganglionares da retina.
Overactivation of NMDA receptors has been described to trigger neuronal death that occurs in diseases such as glaucoma. It is possible that the combination of subunits (NR2A-D) activate intracellular signaling pathways that result in death or survival. Our aim was to investigate the involvement of NR2 subunits and Rac1, a member of Rho GTPase family, in retinal neuronal death. Glutamate-induced neuronal death in vitro was reduced after Rac1 inhibition and by NR2B blocking, but not NR2C/D subunits. Similar results were obtained in vivo after NMDA intravitreous injection, although active Rac1 was mainly detected in Müller glia processes, and it was inhibited by NR2B blockade. In addition, TNF-α level after NMDA injection were reduced by NR2B blocking and Rac1. Thus, our results suggest that excitotoxicity via NR2B/NMDA receptors activate Rac1 in Müller glia cells, which in turn controls the TNF-α production that triggers retinal ganglion cell death.
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8

Pokorny, Morgan R. "The role of Y-box binding protein 1 in prostate cancer." Thesis, Queensland University of Technology, 2013. https://eprints.qut.edu.au/65556/1/Morgan_Pokorny_Thesis.pdf.

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This thesis examined the possible role of Y-box binding protein 1 (YBX1) in prostate cancer aggression and spread. Novel roles were uncovered for YBX1 in the regulation of several genes previously implicated in prostate cancer, as well as showing an effect for YBX1 in increasing tumour cell invasion and movement and reciprocal regulation of androgen-regulated gene networks. In addition, it was found that Y-box 1 regulated several other well-known cancer genes implicated in breast and other cancers. The work performed in this thesis has strengthened the foundations for pursuing YBX1 as a possible central target molecule in prostate cancer therapeutics.
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Newcombe, Anthony Richard. "The biochemical role of the small G protein Rac1 in cell signalling pathways : interaction with RhoGDI and the phagocyte NADPH oxidase component, p67'p'h'o'x." Thesis, University College London (University of London), 2000. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.342224.

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10

Arbuckle, Janeen Lynnae. "Identification and characterization of domains in non-core RAG1." Oklahoma City : [s.n.], 2007.

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Книги з теми "Rac1 protein"

1

Bebchuk, Lucian A. Federalism and takeover law: The race to protect managers from takeovers. Cambridge, MA: National Bureau of Economic Research, 1999.

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Martha, Baer, ed. Safe: The race to protect ourselves in a newly dangerous world. New York: HarperCollins, 2005.

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3

Love Canal revisited : race, class, and gender in environmental activism. Lawrence, Kan: University Press of Kansas, 2008.

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4

Alcalde, Emerson. Antifa. Sao Paulo (SP)]: Autonomia Literaria, 2019.

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5

Hutchinson, Earl Ofari. Betrayed: A history of presidential failure to protect Black lives. Boulder, Colo: Westview Press, 1996.

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6

Patterson, Anita Haya. From Emerson to King: Democracy, race, and the politics of protest. New York: Oxford University Press, 1997.

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7

Smith, Laverne Byrd. Poems of indignation: Revisiting 20th century civil rights and Black awareness movements. Richmond, Va: NorthLight Publishing, 2005.

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8

Black protest and the great migration: A brief history with documents. Boston: Bedford/St. Martin's, 2003.

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9

Hīkoi: Forty years of Māori protest. Wellington, N.Z: Huia Publishers, 2004.

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10

Health, United States Congress House Committee on Energy and Commerce Subcommittee on. A review of efforts to protect jockeys and horses in horseracing: Hearing before the Subcommittee on Health of the Committee on Energy and Commerce, House of Representatives, One Hundred Twelfth Congress, second session, April 30, 2012. Washington: U.S. Government Printing Office, 2013.

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Частини книг з теми "Rac1 protein"

1

Buscemi, Nina, Amanda Doherty-Kirby, Mark A. Sussman, Gilles Lajoie, and Jennifer E. Van Eyk. "Proteomic analysis of Rac1 transgenic mice displaying dilated cardiomyopathy reveals an increase in creatine kinase M-chain protein abundance." In Biochemistry of Hypertrophy and Heart Failure, 145–51. Boston, MA: Springer US, 2003. http://dx.doi.org/10.1007/978-1-4419-9238-3_20.

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Stolarov, Javor P., and Michael H. Wigler. "in vivo Analysis of C-Raf1 — 14-3-3 Interaction." In Interacting Protein Domains, 143–46. Berlin, Heidelberg: Springer Berlin Heidelberg, 1997. http://dx.doi.org/10.1007/978-3-642-60848-3_22.

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3

Sorek, Nadav, and Shaul Yalovsky. "Protein–Lipid Modifications and Targeting of ROP/RAC and Heterotrimeric G Proteins." In Integrated G Proteins Signaling in Plants, 71–90. Berlin, Heidelberg: Springer Berlin Heidelberg, 2009. http://dx.doi.org/10.1007/978-3-642-03524-1_4.

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Lumry, Rufus. "Protein Conformations, “Rack” Mechanisms and Water." In Advances in Chemical Physics, 567–80. Hoboken, NJ, USA: John Wiley & Sons, Inc., 2007. http://dx.doi.org/10.1002/9780470143698.ch39.

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Harvey, Robert S., and Susan Gonzowitz. "Just Another Day: Talk About the Everydayness of Race." In Teaching as Protest, 113–26. New York: Routledge, 2022. http://dx.doi.org/10.4324/9781003183365-6.

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Nieman, David C. "Ultra Marathon Race Competition and Immune Function." In Heat Shock Proteins and Whole Body Physiology, 267–83. Dordrecht: Springer Netherlands, 2009. http://dx.doi.org/10.1007/978-90-481-3381-9_16.

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Malmström, Bo G. "Rack-induced bonding in blue-copper proteins." In EJB Reviews 1994, 157–64. Berlin, Heidelberg: Springer Berlin Heidelberg, 1994. http://dx.doi.org/10.1007/978-3-642-79502-2_12.

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Minto, John. "Race, Rugby and Political Protest in New Zealand: A Personal Account." In Sport, Protest and Globalisation, 131–50. London: Palgrave Macmillan UK, 2016. http://dx.doi.org/10.1057/978-1-137-46492-7_7.

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Wu, Hen-ming, Christine Y. Chen, Qiaohong Duan, Daniel Kita, Candida Nibau, Li-zhen Tao, Yan-jiao Zou, and Alice Y. Cheung. "RAC/ROP GTPases in the Regulation of Polarity and Polar Cell Growth." In Integrated G Proteins Signaling in Plants, 105–22. Berlin, Heidelberg: Springer Berlin Heidelberg, 2009. http://dx.doi.org/10.1007/978-3-642-03524-1_6.

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Fasano, Ottavio, Jean Bernard Crechet, Emmanuele De Vendittis, Regina Zahn, Georg Feger, Alessandra Vitelli, and Andrea Parmeggiani. "Mutations at the RAS2 Locus that, in a ras1- Background, Impair the Growth of Yeast on Nonfermentable Carbon Sources." In The Guanine — Nucleotide Binding Proteins, 251–56. Boston, MA: Springer US, 1989. http://dx.doi.org/10.1007/978-1-4757-2037-2_24.

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Тези доповідей конференцій з теми "Rac1 protein"

1

Rotenberg, Susan A., Xin Zhao, and Shatarupa De. "Abstract 57: Substrates of protein kinase C drive cell rac1-dependent motility." In Proceedings: AACR 106th Annual Meeting 2015; April 18-22, 2015; Philadelphia, PA. American Association for Cancer Research, 2015. http://dx.doi.org/10.1158/1538-7445.am2015-57.

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Hua, Kuo-Tai, Ching-Ting Tan, Chi-Kuan Chen, Min-Wei Chen, Michael Hsiao та Min-Liang Kuo. "Abstract 3422: N-α-acetyltransferase 10 protein suppresses cancer cell metastasis by binding PIX proteins and inhibiting Cdc42/Rac1 activity". У Proceedings: AACR 103rd Annual Meeting 2012‐‐ Mar 31‐Apr 4, 2012; Chicago, IL. American Association for Cancer Research, 2012. http://dx.doi.org/10.1158/1538-7445.am2012-3422.

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Kang, Jia, Rathiga Velaithan, Jayshree L. Hirpara, Thomas Loh, Catherine Brenner, Marie-Veronique Clement, and Shazib Pervaiz. "Abstract 1025: Rac1 is a novel binding partner of the anti-apoptotic protein Bcl-2." In Proceedings: AACR 101st Annual Meeting 2010‐‐ Apr 17‐21, 2010; Washington, DC. American Association for Cancer Research, 2010. http://dx.doi.org/10.1158/1538-7445.am10-1025.

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Sosa‐García, Bernadette, Viviana Vázquez‐Rivera, Julie Dutil, William D. Cress, and Pedro Santiago‐Cardona. "Abstract A68: A functional link between the retinoblastoma protein and the small Rho GTPase Rac1." In Abstracts: First AACR International Conference on Frontiers in Basic Cancer Research--Oct 8–11, 2009; Boston MA. American Association for Cancer Research, 2009. http://dx.doi.org/10.1158/0008-5472.fbcr09-a68.

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Hua, Kuo-Tai, Michael Hsiao, and Min-Liang Kuo. "Abstract 3376: Human arrest defective 1 protein suppresses cancer cell metastasis by binding PIX/Cool proteins and inhibiting Cdc42/Rac1 activity." In Proceedings: AACR 101st Annual Meeting 2010‐‐ Apr 17‐21, 2010; Washington, DC. American Association for Cancer Research, 2010. http://dx.doi.org/10.1158/1538-7445.am10-3376.

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Ajumal, Shani, Jia Kang, Thomas Joseph, Shazib Pervaiz, and Kunchithapadam Swaminathan. "Abstract 3097: Structural and biophysical characterization of anti-apoptotic protein Bcl-2 and GTPase Rac1 interaction." In Proceedings: AACR 107th Annual Meeting 2016; April 16-20, 2016; New Orleans, LA. American Association for Cancer Research, 2016. http://dx.doi.org/10.1158/1538-7445.am2016-3097.

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Tanaka, T., M. Terada, K. Morimoto, and K. Ariyoshi. "MCP-1/CCL2 (Monocyte Chemoattractant Protein-1/CC Chemokine Ligand-2) Enhance Apoptotic Cells Removal (Efferocytosis) through Activation of Rac1." In American Thoracic Society 2009 International Conference, May 15-20, 2009 • San Diego, California. American Thoracic Society, 2009. http://dx.doi.org/10.1164/ajrccm-conference.2009.179.1_meetingabstracts.a3724.

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Palanivel, Chitra, Bailey M. Gabler, Ying Yan, Surinder K. Batra, and Michel M. Ouellette. "Abstract A32: The small GTPase Rac1 controls the stability of Yes-Associated Protein (YAP) independently of the LATS1/2 kinases." In Abstracts: AACR Special Conference on the Hippo Pathway: Signaling, Cancer, and Beyond; May 8-11, 2019; San Diego, CA. American Association for Cancer Research, 2020. http://dx.doi.org/10.1158/1557-3125.hippo19-a32.

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Palanivel, Chitra, Bailey Gabler, Ying Yan, Surinder K. Batra та Michel M. Ouellette. "Abstract 4366: The small GTPase Rac1 controls the stability of the Yes-associated protein 1 (YAP1) independently of the LATS1/2 kinases and SCF-βTRCP ubiquitin ligase". У Proceedings: AACR Annual Meeting 2018; April 14-18, 2018; Chicago, IL. American Association for Cancer Research, 2018. http://dx.doi.org/10.1158/1538-7445.am2018-4366.

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DeLucas, Lawrence J., William Crysel, Terry Bray, Marianna M. Long, Karen M. Moore, and Lance Weise. "Protein Crystal Growth in Space, Past and Future." In ASME 2001 International Mechanical Engineering Congress and Exposition. American Society of Mechanical Engineers, 2001. http://dx.doi.org/10.1115/imece2001/ts-23407.

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Abstract The Center for Biophysical Sciences and Engineering (CBSE) at the University of Alabama at Birmingham has performed protein crystal growth experiments on more than 37 U.S. space shuttle missions. Results from these experiments have clearly demonstrated that the microgravity environment is beneficial in that a number of proteins crystallized were larger and of higher quality than their earth-grown counterparts. Improvement in crystal quality is judged by analysis of ultimate diffraction resolution, individual peak mosaicity, and electron density maps. There are now a number of protein crystals that exhibited resolution improvements of 0.5Å to 1.5Å. Mosaicity studies revealed dramatic decreases in peak widths for the microgravity-grown crystals. These microgravity results plus data from a variety of other investigators have stimulated various space agencies to support fundamental studies in macromolecular crystal growth processes. The CBSE has devoted substantial effort toward the development of dynamically-controlled crystal growth systems which allow scientists to optimize crystallization parameters on Earth or in space. These systems enable monitoring and control of the approach to nucleation and post-nucleation growth phases, thereby dramatically improving the crystal size and x-ray diffraction characteristics. The CBSE is currently designing a complete crystallographic laboratory for the International Space Station including: a crystal growth rack, which will support a variety of crystallization hardware systems; an x-ray diffraction rack for crystal characterization or a complete x-ray data set collection; and robotically-controlled crystal harvesting/cryopreservation systems that can be operated with minimal crew time via telerobotic and/or robotic procedures. Key elements of the x-ray system include unique x-ray focusing technology combined with a lightweight, low power source. The x-ray detection system is based on commercial CCD-based technology. This paper will describe the x-ray facility envisioned for the International Space Station.
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Звіти організацій з теми "Rac1 protein"

1

Delmer, Deborah P., Douglas Johnson, and Alex Levine. The Role of Small Signal Transducing Gtpases in the Regulation of Cell Wall Deposition Patterns in Plants. United States Department of Agriculture, August 1995. http://dx.doi.org/10.32747/1995.7570571.bard.

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The combined research of the groups of Delmer, Levine and Johnson has led to a number of interesting findings with respect to the function of the small GTPase Rac in plants and also opened up new leads for future research. The results have shown: 1) The Rac13 protein undergoes geranylgeranlyation and is also translocated to the plasma membrane as found for Rac in mammals; 2) When cotton Rac13 is highly- expressed in yeast, it leads to an aberrant phenotype reminiscent of mutants impaired in actin function, supporting a role for Rac13 in cytoskeletal organization; 3) From our searches, there is no strong evidence that plants contain homologs of the related CDC42 genes found in yeast and mammals; 4) We have identified a rather unique Rac gene in Arabidopsis that has unusual extensions at both the N- and C-terminal portions of the protein; 5) New evidence was obtained that an oxidative burst characterized by substantial and sustained production of H202 occurs coincident with the onset of secondary wall synthesis in cotton fibers. Further work indicates that the H202 produced may be a signal for the onset of this phase of development and also strongly suggests that Rac plays an important role in signaling for event. Since the secondary walls of plants that contain high levels of lignin and cellulose are the major source of biomass on earth, understanding what signals control this process may well in the future have important implications for manipulating the timing and extent of secondary wall deposition. 6) When the cotton Rac13 promoter is fused to the reporter gene GUS, expression patterns in Arabidopsis indicate very strong and specific expression in developing trichomes and in developing xyelm. Since both of these cell types are engaged in secondary wall synthesis, this further supports a role for Rac in signaling for onset of this process. Since cotton fibers are anatomically defined as trichomes, these data may also be quite useful for future studies in which the trichomes of Arabidopsis may serve as a model for cotton fiber development; the Rac promoter can therefore be useful to drive expression of other genes proposed to affect fiber development and study the effects on the process; 7) The Rac promoter has also been shown to be the best so far tested for use in development of a system for transient transformation of developing cotton fibers, a technique that should have many applications in the field of cotton biotechnology; 8) One candidate protein that may interact with Rac13 to be characterized further in the future is a protein kinase that may be analogous to the PAK kinase that is known to interact with Rac in mammals.
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Yalovsky, Shaul, and Julian Schroeder. The function of protein farnesylation in early events of ABA signal transduction in stomatal guard cells of Arabidopsis. United States Department of Agriculture, January 2002. http://dx.doi.org/10.32747/2002.7695873.bard.

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Loss of function mutations in the farnesyltransferase β subunit gene ERA1 (enhanced response to abscisic acid), cause abscisic acid hypersensitivity in seedlings and in guard cells. This results in slowed water loss of plants in response to drought. Farnesyltransferase (PFT) catalyses the attachment of the 15-carbon isoprenoid farnesyl to conserved cysteine residues located in a conserved C-terminal domain designated CaaX box. PFT is a heterodimeric protein comprised of an a and b sununits. The a subunit is shared between PFT and geranylgeranyltransferase-I (PGGTI) which catalyses the attachemt of the 20-carbon isoprenoid geranylgeranyl to CaaX box proteins in which the last amino acid is almost always leucine and in addition have a polybasic domain proximal to the CaaL box. Preliminary data presented in the proposal showed that increased cytoplasmic Ca2+ concentration in stomal guard cells in response to non-inductive ABA treatements. The goals set in the proposal were to characterize better how PFT (ERA1) affects ABA induced Ca2+ concentrations in guard cells and to identify putative CaaX box proteins which function as negative regulators of ABA signaling and which function is compromised in era1 mutant plants. To achieve these goals we proposed to use camelion Ca2+ sensor protein, high throughput genomic to identify the guard cell transcriptome and test prenylation of candidate proteins. We also proposed to focus our efforts of RAC small GTPases which are prenylated proteins which function in signaling. Our results show that farnesyltransferaseprenylates protein/s that act between the points of ABA perception and the activation of plasma membrane calcium influx channels. A RAC protein designated AtRAC8/AtRop10 also acts in negative regulation of ABA signaling. However, we discovered that this protein is palmitoylated and not prenylated although it contains a C-terminal CXXX motif. We further discovered a unique C-terminal sequence motif required for membrane targeting of palmitoylatedRACs and showed that their function is prenylation independent. A GC/MS based method for expression in plants, purification and analysis of prenyl group was developed. This method would allow highly reliable identification of prenylated protein. Mutants in the shared α subunit of PFT and PGGT-I was identified and characterized and was shown to be ABA hypersensitive but less than era1. This suggested that PFT and PGGT-I have opposing functions in ABA signaling. Our results enhanced the understanding of the role of protein prenylation in ABA signaling and drought resistance in plants with the implications of developing drought resistant plants. The results of our studies were published 4 papers which acknowledge support from BARD.
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3

Watad, Abed A., Paul Michael Hasegawa, Ray A. Bressan, Alexander Vainstein, and Yigal Elad. Osmotin and Osmotin-Like Proteins as a Novel Source for Phytopathogenic Fungal Resistance in Transgenic Carnation and Tomato Plants. United States Department of Agriculture, January 2000. http://dx.doi.org/10.32747/2000.7573992.bard.

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The goal of this project is to enhance fungal resistance of carnation and tomato through the ectopic expression of osmotin and other pathogenesis-related (PR) proteins. The research objectives were to evaluate in vitro antifungal activity of osmotin and osmotin and other PR protein combinations against phytopathogens (including Fusarium oxysporum, Verticillium dahliae, Botrytus cinerea or Phytophthora infestans), develop protocols for efficient transformation of carnation and tomato, express PR proteins in transgenic carnation and tomato and evaluate fungal resistance of transgenic plants. Protocols for microprojectile bombardment and Agrobacterium-mediated transformation of carnation were developed that are applicable for the biotechnology of numerous commercial cultivars. Research established an efficient organogenetic regeneration system, optimized gene delivery and transgene expression and defined parameters requisite to the high frequency recovery of transgenic plants. Additionally, an efficient Agrobacterium-mediated transformation protocol was developed for tomato that is applicable for use with numerous commercial varieties. Rigorous selection and reducing the cytokinin level in medium immediately after shoot induction resulted in substantially greater frequency of adventitious shoots that developed defined stems suitable for rooting and reconstitution of transgenic plants. Transformation vectors were constructed for co-expression of genes encoding osmotin and tobacco chitinase Ia or PR-1b. Expression of osmotin, PR-1 and/or chitinase in transgenic carnation mediated a high level resistance of cv. White Sim (susceptible variety) to F. oxysporum f. sp. dianthi, race 2 in greenhouse assays. These plants are being evaluated in field tests. Comprehensive analysis (12 to 17 experiments) indicated that germination of B. cinerea conidia was unaffected by PR protein expression but germ tube elongation was reduced substantially. The disease severity was significantly attenuated by PR protein expression. Constitutive expression of osmotin in transgenic tomato increased resistance to B. cinerea, and P. infestans. Grey mold and late blight resistance was stable through the third selfed generation. The research accomplished in this project will have profound effects on the use of biotechnology to improve carnation and tomato. Transformation protocols that are applicable for efficient stable gene transfer to numerous commercial varieties of carnation and tomato are the foundation for the capacity to bioengineer these crops. The research further establishes that PR proteins provide a measure of enhanced disease resistance. However, considerations of PR protein combinations and conditional regulation and targeting are likely required to achieve; sustained level of resistance.
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4

Spiegel, Yitzhak, Michael McClure, Itzhak Kahane, and B. M. Zuckerman. Characterization of the Phytophagous Nematode Surface Coat to Provide New Strategies for Biocontrol. United States Department of Agriculture, November 1995. http://dx.doi.org/10.32747/1995.7613015.bard.

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Chemical composition and biological role of the surface coat (SC) of the root-knot nematodes, Meloidogyne spp. are described. SC proteins of M. incognita race 3 infective juveniles (J2) were characterized by electrophoresis and western blotting of extracts from radioiodine and biotin-labelled nematodes. J2 labelled with radioiodine and biotin released 125I and biotin-labelled molecules into water after 20 hours incubation, indicating that SC proteins may be loosely attached to the nematode. Antiserum to the principal protein reacted with the surface of live J2 and with surface proteins previously separated by electrophoresis. Human red blood cells (HRBC) adhered to J2 of several tylenchid nematodes over the entire nematode body. HRBC adhered also to nylon fibers coated with SC extracted from M. javanica J2; binding was Ca++/Mg++ dependent, and decreased when the nylon fibers were coated with bovine serum albumin, or pre-incubated with fucose and mannose. These experiments support a working hypothesis that RBC adhesion involves carbohydrate moieties of HRBC and carbohydrate-recognition domain(s) (CRD) distributed on the nematode surface. To our knowledge, this is the first report of a surface CRD i the phylum Nematoda. Gold-conjugated lectins and neoglycoproteins combined with silver enhancement have been used for the detection of carbohydrates and CRD, respectively, on the SC of M. javanica J2. Biotin reagents were used to trace surface proteins, specifically, on live J2. The labile and transitory nature of the SC was demonstrated by the dynamics of HRBC adherence to detergent-treated J2, J2 at different ages or fresh-hatched J2 held at various temperatures. SC recovery was demonstrated also by a SDS-PAGE profile. Monoclonal antibodies developed to a cuticular protein of M. incognita J2 gave a slight, but significant reduction in attachment of Pasteuria penetrans spores. Spore attachment as affected by several enzymes was inconsistent: alcian blue, which specifically blocks sulfyl groups, had no afffect on spore attachment. Treatment with cationized ferritin alone or catonized ferritin following monoclonal antibody caused significant decreases in spore attachment. Those results suggest a role in attachment by negatively charged groups.
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5

Bebchuk, Lucian Arye, and Allen Ferrell. Federalism and Takeover Law: The Race to Protect Managers from Takeovers. Cambridge, MA: National Bureau of Economic Research, June 2000. http://dx.doi.org/10.3386/w7232.

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6

Smith, Margaret, Nurit Katzir, Susan McCouch, and Yaakov Tadmor. Discovery and Transfer of Genes from Wild Zea Germplasm to Improve Grain Oil and Protein Composition of Temperate Maize. United States Department of Agriculture, 1998. http://dx.doi.org/10.32747/1998.7580683.bard.

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Project Objectives 1. Develop and amplify two interspecific populations (annual and perennial teosintes x elite maize inbred) as the basis for genetic analysis of grain quality. 2. Identify quantitative trait loci (QTLs) from teosinte that improve oil, protein, and essential amino acid composition of maize grain. 3. Develop near isogenic lines (NILs) to quantify QTL contributions to grain quality and as a resource for future breeding and gene cloning efforts. 4. Analyze the contribution of these QTLs to hybrid performance in both the US and Israel. 5. Measure the yield potential of improved grain quality hybrids. (NOTE: Yield potential could not be evaluated due to environmentally-caused failure of the breeding nursery where seed was produced for this evaluation.) Background: Maize is a significant agricultural commodity worldwide. As an open pollinated crop, variation within the species is large and, in most cases, sufficient to supply the demand for modem varieties and for new environments. In recent years there is a growing demand for maize varieties with special quality attributes. While domesticated sources of genetic variation for high oil and protein content are limited, useful alleles for these traits may remain in maize's wild relative, teosinte. We utilized advanced backcross (AB) analysis to search for QTLs contributing to oil and protein content from two teosinte accessions: Zea mays ssp. mexicana Race Chalco, an annual teosinte (referred to as Chalco), and Z diploperennis Race San Miguel, a perennial teosinte (referred to as Diplo). Major Conclusions and Achievements Two NILs targeting a Diplo introgression in bin 1.04 showed a significant increase in oil content in homozygous sib-pollinated seed when compared to sibbed seed of their counterpart non-introgressed controls. These BC4S2 NILs, referred to as D-RD29 and D-RD30, carry the Diplo allele in bin 1.04 and the introgression extends partially into bins 1.03 and 1.05. These NILs remain heterozygous in bins 4.01 and 8.02, but otherwise are homozygous for the recurrent parent (RD6502) alleles. NILs were developed also for the Chalco introgression in bin 1.04 but these do not show any improvement in oil content, suggesting that the Chalco alleles differ from the Diplo alleles in this region. Testcross Fl seed and sibbed grain from these Fl plants did not show any effect on oil content from this introgression, suggesting that it would need to be present in both parents of a maize hybrid to have an effect on oil content. Implications, both Scientific and Agricultural The Diplo region identified increases oil content by 12.5% (from 4.8% to 5.4% oil in the seed). Although this absolute difference is not large in agronomic terms, this locus could provide additive increases to oil content in combination with other maize-derived loci for high oil. To our knowledge, this is the first confirmed report of a QTL from teosinte for improved grain oil content in maize. It suggests that further research on grain quality alleles from maize wild relatives would be of both scientific and agricultural interest.
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7

Smith, Margaret, Nurit Katzir, Susan McCouch, and Yaakov Tadmor. Discovery and Transfer of Genes from Wild Zea Germplasm to Improve Grain Oil and Protein Composition of Temperate Maize. United States Department of Agriculture, October 2002. http://dx.doi.org/10.32747/2002.7695846.bard.

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Анотація:
Project Objectives 1. Develop and amplify two interspecific populations (annual and perennial teosintes x elite maize inbred) as the basis for genetic analysis of grain quality. 2. Identify quantitative trait loci (QTLs) from teosinte that improve oil, protein, and essential amino acid composition of maize grain. 3. Develop near isogenic lines (NILs) to quantify QTL contributions to grain quality and as a resource for future breeding and gene cloning efforts. 4. Analyze the contribution of these QTLs to hybrid performance in both the US and Israel. 5. Measure the yield potential of improved grain quality hybrids. (NOTE: Yield potential could not be evaluated due to environmentally-caused failure of the breeding nursery where seed was produced for this evaluation.) Background: Maize is a significant agricultural commodity worldwide. As an open pollinated crop, variation within the species is large and, in most cases, sufficient to supply the demand for modem varieties and for new environments. In recent years there is a growing demand for maize varieties with special quality attributes. While domesticated sources of genetic variation for high oil and protein content are limited, useful alleles for these traits may remain in maize's wild relative, teosinte. We utilized advanced backcross (AB) analysis to search for QTLs contributing to oil and protein content from two teosinte accessions: Zea mays ssp. mexicana Race Chalco, an annual teosinte (referred to as Chalco), and Z diploperennis Race San Miguel, a perennial teosinte (referred to as Diplo). Major Conclusions and Achievements Two NILs targeting a Diplo introgression in bin 1.04 showed a significant increase in oil content in homozygous sib-pollinated seed when compared to sibbed seed of their counterpart non-introgressed controls. These BC4S2 NILs, referred to as D-RD29 and D-RD30, carry the Diplo allele in bin 1.04 and the introgression extends partially into bins 1.03 and 1.05. These NILs remain heterozygous in bins 4.01 and 8.02, but otherwise are homozygous for the recurrent parent (RD6502) alleles. NILs were developed also for the Chalco introgression in bin 1.04 but these do not show any improvement in oil content, suggesting that the Chalco alleles differ from the Diplo alleles in this region. Testcross Fl seed and sibbed grain from these Fl plants did not show any effect on oil content from this introgression, suggesting that it would need to be present in both parents of a maize hybrid to have an effect on oil content. Implications, both Scientific and Agricultural The Diplo region identified increases oil content by 12.5% (from 4.8% to 5.4% oil in the seed). Although this absolute difference is not large in agronomic terms, this locus could provide additive increases to oil content in combination with other maize-derived loci for high oil. To our knowledge, this is the first confirmed report of a QTL from teosinte for improved grain oil content in maize. It suggests that further research on grain quality alleles from maize wild relatives would be of both scientific and agricultural interest.
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8

Dubcovsky, Jorge, Tzion Fahima, and Ann Blechl. Molecular characterization and deployment of the high-temperature adult plant stripe rust resistance gene Yr36 from wheat. United States Department of Agriculture, November 2013. http://dx.doi.org/10.32747/2013.7699860.bard.

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Stripe rust, caused by Puccinia striiformis f. sp. tritici is one of the most destructive fungal diseases of wheat. Virulent races that appeared within the last decade caused drastic cuts in yields. The incorporation of genetic resistance against this pathogen is the most cost-effective and environmentally friendly solution to this problem. However, race specific seedling resistance genes provide only a temporary solution because fungal populations rapidly evolve to overcome this type of resistance. In contrast, high temperature adult plant (HTAP) resistance genes provide a broad spectrum resistance that is partial and more durable. The cloning of the first wheat HTAP stripe rust resistance gene Yr36 (Science 2009, 323:1357), funded by our previous (2007-2010) BARD grant, provided us for the first time with an entry point for understanding the mechanism of broad spectrum resistance. Two paralogous copies of this gene are tightly linked at the Yr36 locus (WKS1 and WKS2). The main objectives of the current study were to characterize the Yr36 (WKS) resistance mechanism and to identify and characterize alternative WKSgenes in wheat and wild relatives. We report here that the protein coded by Yr36, designated WKS1, that has a novel architecture with a functional kinase and a lipid binding START domain, is localized to chloroplast. Our results suggest that the presence of the START domain may affect the kinase activity. We have found that the WKS1 was over-expressed on leaf necrosis in wheat transgenic plants. When the isolated WKS1.1 splice variant transcript was transformed into susceptible wheat it conferred resistance to stripe rust, but the truncated variant WKS1.2 did not confer resistance. WKS1.1 and WKS1.2 showed different lipid binding profiling. WKS1.1 enters the chloroplast membrane, while WKS1.2 is only attached outside of the chloroplast membrane. The ascorbate peroxidase (APX) activity of the recombinant protein of TmtAPXwas found to be reduced by WKS1.1 protein in vitro. The WKS1.1 mature protein in the chloroplast is able to phosphorylate TmtAPXprotein in vivo. WKS1.1 induced cell death by suppressing APX activity and reducing the ability of the cell to detoxify reactive oxygen. The decrease of APX activity reduces the ability of the plant to detoxify the reactive H2O2 and is the possible mechanism underlying the accelerated cell death observed in the transgenic plants overexpressing WKS1.1 and in the regions surrounding a stripe rust infection in the wheat plants carrying the natural WKS1.1 gene. WKS2 is a nonfunctional paralog of WKS1 in wild emmer wheat, probably due to a retrotransposon insertion close to the alternative splicing site. In some other wild relatives of wheat, such as Aegilops comosa, there is only one copy of this gene, highly similar to WKS2, which is lucking the retrotransposon insertion. WKS2 gene present in wheat and WKS2-Ae from A. showed a different pattern of alternative splice variants, regardless of the presence of the retrotransposon insertion. Susceptible Bobwhite transformed with WKS2-Ae (without retrotansposon insertion in intron10), which derived from Aegilops comosaconferred resistance to stripe rust in wheat. The expression of WKS2-Ae in transgenic plants is up-regulated by temperature and pathogen infection. Combination of WKS1 and WKS2-Ae shows improved stripe rust resistance in WKS1×WKS2-Ae F1 hybrid plants. The obtained results show that WKS1 protein is accelerating programmed cell death observed in the regions surrounding a stripe rust infection in the wheat plants carrying the natural or transgenic WKS1 gene. Furthermore, characterization of the epistatic interactions of Yr36 and Yr18 demonstrated that these two genes have additive effects and can therefore be combined to increase partial resistance to this devastating pathogen of wheat. These achievements may have a broad impact on wheat breeding efforts attempting to protect wheat yields against one of the most devastating wheat pathogen.
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9

Joel, Daniel M., Steven J. Knapp, and Yaakov Tadmor. Genomic Approaches for Understanding Virulence and Resistance in the Sunflower-Orobanche Host-Parasite Interaction. United States Department of Agriculture, August 2011. http://dx.doi.org/10.32747/2011.7592655.bard.

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Oroginal Objectives: (i) identify DNA markers linked to the avirulence (Avr) locus and locate the Avr locus through genetic mapping with an inter-race Orobanche cumana population; (ii) develop high-throughput fingerprint DNA markers for genotypingO. cumana races; (iii) identify nucleotide binding domain leucine rich repeat (NB-LRR) genes encoding R proteins conferring resistance to O. cumana in sunflower; (iv) increase the resolution of the chromosomal segment harboring Or₅ and related R genes through genetic and physical mapping in previously and newly developed mapping populations of sunflower; and (v) develop high-throughput DNA markers for rapidly and efficiently identifying and transferring sunflower R genes through marker-assisted selection. Revisions made during the course of project: Following changes in O. cumana race distribution in Israel, the newly arrived virulent race H was chosen for further analysis. HA412-HO, which was primarily chosen as a susceptible sunflower cultivar, was more resistant to the new parasite populations than var. Shemesh, thus we shifted sunflower research into analyzing the resistance of HA412-HO. We exceeded the deliverables for Objectives #3-5 by securing funding for complete physical and high-density genetic mapping of the sunflower genome, in addition to producing a complete draft sequence of the sunflower genome. We discovered limited diversity between the parents of the O. cumana population developed for the mapping study. Hence, the developed DNA marker resources were insufficient to support genetic map construction. This objective was beyond the scale and scope of the funding. This objective is challenging enough to be the entire focus of follow up studies. Background to the topic: O. cumana, an obligate parasitic weed, is one of the most economically important and damaging diseases of sunflower, causes significant yield losses in susceptible genotypes, and threatens production in Israel and many other countries. Breeding for resistance has been crucial for protecting sunflower from O. cumana, and problematic because new races of the pathogen continually emerge, necessitating discovery and deployment of new R genes. The process is challenging because of the uncertainty in identifying races in a genetically diverse parasite. Major conclusions, solutions, achievements: We developed a small collection of SSR markers for genetic mapping in O. cumana and completed a diversity study to lay the ground for objective #1. Because DNA sequencing and SNPgenotyping technology dramatically advanced during the course of the study, we recommend shifting future work to SNP discovery and mapping using array-based approaches, instead of SSR markers. We completed a pilot study using a 96-SNP array, but it was not large enough to support genetic mapping in O.cumana. The development of further SNPs was beyond the scope of the grant. However, the collection of SSR markers was ideal for genetic diversity analysis, which indicated that O. cumanapopulations in Israel considerably differ frompopulations in other Mediterranean countries. We supplied physical and genetic mapping resources for identifying R-genes in sunflower responsible for resistance to O. cumana. Several thousand mapped SNP markers and a complete draft of the sunflower genome sequence are powerful tools for identifying additional candidate genes and understanding the genomic architecture of O. cumana-resistanceanddisease-resistance genes. Implications: The OrobancheSSR markers have utility in sunflower breeding and genetics programs, as well as a tool for understanding the heterogeneity of races in the field and for geographically mapping of pathotypes.The segregating populations of both Orobanche and sunflower hybrids are now available for QTL analyses.
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10

Woods, Rachel, Alison Zhong, and Madelyn Vincent. Factors Associated with Influenza & Tdap Vaccine Uptake in Pregnant Patients at the UT Family Medicine Clinic in Memphis. University of Tennessee Health Science Center, 2021. http://dx.doi.org/10.21007/com.lsp.2020.0003.

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INTRODUCTION: Given the increased risk for infections among pregnant patients and newborns, vaccination against influenza (>50,000,000 annual US cases affecting all ages) and pertussis (>15,000 annual US cases disproportionately affecting newborns) are recommended among pregnant patients in order to protect them and their babies via passive immunity to cover a newborn’s window of vaccine ineligibility. Though flu and Tdap vaccination rates among pregnant patients have been trending upwards nationally, there is still room for improvement to achieve optimal rates. OBJECTIVES: The primary objectives were to study factors that affect the vaccination rates at the University of Tennessee Family Medicine Clinic at Memphis (UTFMC-M), compare those rates with national pregnancy flu/Tdap vaccination rates, and to generate recommendations based off observed factors associated with vaccine uptake to improve flu/Tdap vaccination rates in UTFMC-M pregnant patients. METHODS: This was a retrospective chart review of UTFMC-M patients who were pregnant from September 1, 2019-April 24, 2020 (included 2019-2020 flu season) (n=465). Variables studied included demographic data (race, age, insurance), immunization history (vaccine status, history of physician encouragement), and prenatal history (parity, number of prenatal visits, trimester at first visit, high risk clinic (HRC) admittance status). Vaccination status was based on ACIP recommendations (Flu shot eligible = any gestational age; Tdap eligible = ≥27 weeks). Positive HRC admittance was noted for patients with ≥2 visits to the UTFMC-M HRC, a clinic that specializes in high risk pregnant patient care. RESULTS: The patient sample was predominantly black (84.3%) and insured by Medicaid programs (88%). Among eligible UTFMC-M pregnant patients, 50.1% were flu-vaccinated (n=465); 73.8% were Tdap-vaccinated (n=317); and 52.1% were Flu+Tdap-vaccinated (n=317). No significant associations were found between vaccine uptake and HRC status, parity, and age. However, statistically significant relationships were found between vaccine uptake and physician encouragement (positive relationship with flu shot: X2(1, N = 465) =131, p < 0.001, Tdap: X2 (6, N = 465) =476, p < 0.001), number of prenatal visits (flu shot group median 8 visits, Tdap group median 9 visits vs. unvaccinated group median 4 visits; p < 0.001), and early trimester age at first prenatal visit (X2(6, N = 465) =47.635 , p CONCLUSION: 2019-2020 UTFMC-M vaccination rates were on par with 2018-2019 US flu vaccine rates and higher than 2018-2019 US Tdap and Flu+Tdap rates. There were statistically significant relationships between vaccine uptake at UTFMC-M and physician encouragement, number of prenatal visits, and early trimester age at first prenatal visit but no significant relationships with UTFMC-M HRC admittance, parity, or age. Recommendations following from our observations to address further vaccine rate improvement include: continue vaccine encouragement, continue booking multiple visits (8 for flu, 9 for Tdap), prioritize Tdap vaccine higher for late trimester intake patients, and focus on flu vaccine encouragement and education.
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