Literatura académica sobre el tema "CDC50 proteins"
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Artículos de revistas sobre el tema "CDC50 proteins"
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Furuta, Nobumichi, Konomi Fujimura-Kamada, Koji Saito, Takaharu Yamamoto y Kazuma Tanaka. "Endocytic Recycling in Yeast Is Regulated by Putative Phospholipid Translocases and the Ypt31p/32p–Rcy1p Pathway". Molecular Biology of the Cell 18, n.º 1 (enero de 2007): 295–312. http://dx.doi.org/10.1091/mbc.e06-05-0461.
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Phospholipid translocases (PLTs) have been implicated in the generation of phospholipid asymmetry in membrane bilayers. In budding yeast, putative PLTs are encoded by the DRS2 gene family of type 4 P-type ATPases. The homologous proteins Cdc50p, Lem3p, and Crf1p are potential noncatalytic subunits of Drs2p, Dnf1p and Dnf2p, and Dnf3p, respectively; these putative heteromeric PLTs share an essential function for cell growth. We constructed temperature-sensitive mutants of CDC50 in the lem3Δ crf1Δ background (cdc50-ts mutants). Screening for multicopy suppressors of cdc50-ts identified YPT31/32, two genes that encode Rab family small GTPases that are involved in both the exocytic and endocytic recycling pathways. The cdc50-ts mutants did not exhibit major defects in the exocytic pathways, but they did exhibit those in endocytic recycling; large membranous structures containing the vesicle-soluble N-ethylmaleimide-sensitive factor attachment protein receptor Snc1p intracellularly accumulated in these mutants. Genetic results suggested that the YPT31/32 effector RCY1 and CDC50 function in the same signaling pathway, and simultaneous overexpression of CDC50, DRS2, and GFP-SNC1 restored growth as well as the plasma membrane localization of GFP-Snc1p in the rcy1Δ mutant. In addition, Rcy1p coimmunoprecipitated with Cdc50p-Drs2p. We propose that the Ypt31p/32p–Rcy1p pathway regulates putative phospholipid translocases to promote formation of vesicles destined for the trans-Golgi network from early endosomes.
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Saito, Koji, Konomi Fujimura-Kamada, Nobumichi Furuta, Utako Kato, Masato Umeda y Kazuma Tanaka. "Cdc50p, a Protein Required for Polarized Growth, Associates with the Drs2p P-Type ATPase Implicated in Phospholipid Translocation in Saccharomyces cerevisiae". Molecular Biology of the Cell 15, n.º 7 (julio de 2004): 3418–32. http://dx.doi.org/10.1091/mbc.e03-11-0829.
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Cdc50p, a transmembrane protein localized to the late endosome, is required for polarized cell growth in yeast. Genetic studies suggest that CDC50 performs a function similar to DRS2, which encodes a P-type ATPase of the aminophospholipid translocase (APT) subfamily. At low temperatures, drs2Δ mutant cells exhibited depolarization of cortical actin patches and mislocalization of polarity regulators, such as Bni1p and Gic1p, in a manner similar to the cdc50Δ mutant. Both Cdc50p and Drs2p were localized to the trans-Golgi network and late endosome. Cdc50p was coimmunoprecipitated with Drs2p from membrane protein extracts. In cdc50Δ mutant cells, Drs2p resided on the endoplasmic reticulum (ER), whereas Cdc50p was found on the ER membrane in drs2Δ cells, suggesting that the association on the ER membrane is required for transport of the Cdc50p-Drs2p complex to the trans-Golgi network. Lem3/Ros3p, a homolog of Cdc50p, was coimmunoprecipitated with another APT, Dnf1p; Lem3p was required for exit of Dnf1p out of the ER. Both Cdc50p-Drs2p and Lem3p-Dnf1p were confined to the plasma membrane upon blockade of endocytosis, suggesting that these proteins cycle between the exocytic and endocytic pathways, likely performing redundant functions. Thus, phospholipid asymmetry plays an important role in the establishment of cell polarity; the Cdc50p/Lem3p family likely constitute potential subunits specific to unique P-type ATPases of the APT subfamily.
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García-Sánchez, Sebastián, María P. Sánchez-Cañete, Francisco Gamarro y Santiago Castanys. "Functional role of evolutionarily highly conserved residues, N-glycosylation level and domains of the Leishmania miltefosine transporter-Cdc50 subunit". Biochemical Journal 459, n.º 1 (14 de marzo de 2014): 83–94. http://dx.doi.org/10.1042/bj20131318.
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Cdc50 proteins are associated with phospholipid translocating P4-ATPases. Highly conserved and N-glycosylated residues of LiRos3, a Leishmania infantum Cdc50 protein, play a role in correct trafficking to the plasma membrane and in the activity of the miltefosine transporter.
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López-Marqués, Rosa L., Lisbeth R. Poulsen, Susanne Hanisch, Katharina Meffert, Morten J. Buch-Pedersen, Mia K. Jakobsen, Thomas Günther Pomorski y Michael G. Palmgren. "Intracellular Targeting Signals and Lipid Specificity Determinants of the ALA/ALIS P4-ATPase Complex Reside in the Catalytic ALA α-Subunit". Molecular Biology of the Cell 21, n.º 5 (marzo de 2010): 791–801. http://dx.doi.org/10.1091/mbc.e09-08-0656.
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Members of the P4 subfamily of P-type ATPases are believed to catalyze flipping of phospholipids across cellular membranes, in this way contributing to vesicle biogenesis in the secretory and endocytic pathways. P4-ATPases form heteromeric complexes with Cdc50-like proteins, and it has been suggested that these act as β-subunits in the P4-ATPase transport machinery. In this work, we investigated the role of Cdc50-like β-subunits of P4-ATPases for targeting and function of P4-ATPase catalytic α-subunits. We show that the Arabidopsis P4-ATPases ALA2 and ALA3 gain functionality when coexpressed with any of three different ALIS Cdc50-like β-subunits. However, the final cellular destination of P4-ATPases as well as their lipid substrate specificity are independent of the nature of the ALIS β-subunit they were allowed to interact with.
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Hanadate, Yuki, Yumiko Saito-Nakano, Kumiko Nakada-Tsukui y Tomoyoshi Nozaki. "Identification and Characterization of the Entamoeba Histolytica Rab8a Binding Protein: A Cdc50 Homolog". International Journal of Molecular Sciences 19, n.º 12 (30 de noviembre de 2018): 3831. http://dx.doi.org/10.3390/ijms19123831.
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Membrane traffic plays a pivotal role in virulence in the enteric protozoan parasite Entamoeba histolytica. EhRab8A small GTPase is a key regulator of membrane traffic at the endoplasmic reticulum (ER) of this protist and is involved in the transport of plasma membrane proteins. Here we identified the binding proteins of EhRab8A. The Cdc50 homolog, a non-catalytic subunit of lipid flippase, was identified as an EhRab8A binding protein candidate by affinity coimmunoprecipitation. Binding of EhRab8A to EhCdc50 was also confirmed by reciprocal immunoprecipitation and blue-native polyacrylamide gel electrophoresis, the latter of which revealed an 87 kDa complex. Indirect immunofluorescence imaging with and without Triton X100 showed that endogenous EhCdc50 localized on the surface in the absence of permeabilizing agent but was observed on the intracellular structures and overlapped with the ER marker Bip when Triton X100 was used. Overexpression of N-terminal HA-tagged EhCdc50 impaired its translocation to the plasma membrane and caused its accumulation in the ER. As reported previously in other organisms, overexpression and accumulation of Cdc50 in the ER likely inhibited surface transport and function of the plasma membrane lipid flippase P4-ATPase. Interestingly, HA-EhCdc50-expressing trophozoites gained resistance to miltefosine, which is consistent with the prediction that HA-EhCdc50 overexpression caused its accumulation in the ER and mislocalization of the unidentified lipid flippase. Similarly, EhRab8A gene silenced trophozoites showed increased resistance to miltefosine, supporting EhRab8A-dependent transport of EhCdc50. This study demonstrated for the first time that EhRab8A mediates the transport of EhCdc50 and lipid flippase P4-ATPase from the ER to the plasma membrane.
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Bryde, Susanne, Hanka Hennrich, Patricia M. Verhulst, Philippe F. Devaux, Guillaume Lenoir y Joost C. M. Holthuis. "CDC50 Proteins Are Critical Components of the Human Class-1 P4-ATPase Transport Machinery". Journal of Biological Chemistry 285, n.º 52 (20 de octubre de 2010): 40562–72. http://dx.doi.org/10.1074/jbc.m110.139543.
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van der Velden, Lieke M., Catharina G. K. Wichers, Adriana E. D. van Breevoort, Jonathan A. Coleman, Robert S. Molday, Ruud Berger, Leo W. J. Klomp y Stan F. J. van de Graaf. "Heteromeric Interactions Required for Abundance and Subcellular Localization of Human CDC50 Proteins and Class 1 P4-ATPases". Journal of Biological Chemistry 285, n.º 51 (14 de octubre de 2010): 40088–96. http://dx.doi.org/10.1074/jbc.m110.139006.
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Misu, Kenjiro, Konomi Fujimura-Kamada, Takashi Ueda, Akihiko Nakano, Hiroyuki Katoh y Kazuma Tanaka. "Cdc50p, a Conserved Endosomal Membrane Protein, Controls Polarized Growth in Saccharomyces cerevisiae". Molecular Biology of the Cell 14, n.º 2 (febrero de 2003): 730–47. http://dx.doi.org/10.1091/mbc.e02-06-0314.
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During the cell cycle of the yeast Saccharomyces cerevisiae, the actin cytoskeleton and the growth of cell surface are polarized, mediating bud emergence, bud growth, and cytokinesis. We identified CDC50 as a multicopy suppressor of the myo3 myo5-360 temperature-sensitive mutant, which is defective in organization of cortical actin patches. The cdc50 null mutant showed cold-sensitive cell cycle arrest with a small bud as reported previously. Cortical actin patches and Myo5p, which are normally localized to polarization sites, were depolarized in the cdc50 mutant. Furthermore, actin cables disappeared, and Bni1p and Gic1p, effectors of the Cdc42p small GTPase, were mislocalized in the cdc50 mutant. As predicted by its amino acid sequence, Cdc50p appears to be a transmembrane protein because it was solubilized from the membranes by detergent treatment. Cdc50p colocalized with Vps21p in endosomal compartments and was also localized to the class E compartment in thevps27 mutant. The cdc50 mutant showed defects in a late stage of endocytosis but not in the internalization step. It showed, however, only modest defects in vacuolar protein sorting. Our results indicate that Cdc50p is a novel endosomal protein that regulates polarized cell growth.
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Li, Xin, Baohui Chen, Sawako Yoshina, Tanxi Cai, Fuquan Yang, Shohei Mitani y Xiaochen Wang. "Inactivation of Caenorhabditis elegans aminopeptidase DNPP-1 restores endocytic sorting and recycling in tat-1 mutants". Molecular Biology of the Cell 24, n.º 8 (15 de abril de 2013): 1163–75. http://dx.doi.org/10.1091/mbc.e12-10-0730.
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In Caenorhabditis elegans, the P4-ATPase TAT-1 and its chaperone, the Cdc50 family protein CHAT-1, maintain membrane phosphatidylserine (PS) asymmetry, which is required for membrane tubulation during endocytic sorting and recycling. Loss of tat-1 and chat-1 disrupts endocytic sorting, leading to defects in both cargo recycling and degradation. In this study, we identified the C. elegans aspartyl aminopeptidase DNPP-1, loss of which suppresses the sorting and recycling defects in tat-1 mutants without reversing the PS asymmetry defect. We found that tubular membrane structures containing recycling cargoes were restored in dnpp-1 tat-1 double mutants and that these tubules overlap with RME-1–positive recycling endosomes. The restoration of the tubular structures in dnpp-1 tat-1 mutants requires normal functions of RAB-5, RAB-10, and RME-1. In tat-1 mutants, we observed alterations in membrane surface charge and targeting of positively charged proteins that were reversed by loss of dnpp-1. DNPP-1 displays a specific aspartyl aminopeptidase activity in vitro, and its enzymatic activity is required for its function in vivo. Our data reveal the involvement of an aminopeptidase in regulating endocytic sorting and recycling and suggest possible roles of peptide signaling and/or protein metabolism in these processes.
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Park, Chong J., Sukgil Song, Thomas H. Giddings, Hyeon-Su Ro, Krisada Sakchaisri, Jung-Eun Park, Yeon-Sun Seong, Mark Winey y Kyung S. Lee. "Requirement for Bbp1p in the Proper Mitotic Functions of Cdc5p in Saccharomyces cerevisiae". Molecular Biology of the Cell 15, n.º 4 (abril de 2004): 1711–23. http://dx.doi.org/10.1091/mbc.e03-07-0461.
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The polo-box domain of the budding yeast polo kinase Cdc5p plays an essential role for targeting the catalytic activity of Cdc5p to spindle pole bodies (SPBs) and cytokinetic neck-filaments. Here, we report the isolation of Bbp1p as a polo-box interacting protein by a yeast two-hybrid screen. Bbp1p localizes to the periphery of the central plaque of the SPB and plays an important role in SPB duplication. Similarly, Cdc5p localized to the cytoplasmic periphery of the SPB. In vitro binding studies showed that Cdc5p interacted with the N-terminal domain of Bbp1p (Bbp1pΔC), but apparently not with Mps2p, a component shown to form a stable complex with Bbp1p. In addition, Bbp1p, but likely not Mps2p, was required for proper localization of Cdc5p to the SPB. The C-terminal coiled-coil domain of Bbp1p (Bbp1p243–385), which is crucial for both the homodimerization and the SPB localization, could target the localization-defective Cdc5pΔC to the SPB and induce the release of Cdc14p from the nucleolus. Consistent with this observation, expression of CDC5ΔC-BBP1243–385 under CDC5 promoter control partially complemented the cdc5Δ defect. These data suggest that Bbp1pΔC interacts with the polo-box domain of Cdc5p, and this interaction is critical for the subcellular localization and mitotic functions of Cdc5p.
Tesis sobre el tema "CDC50 proteins"
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Lamy, Anaïs. "Lipid Flippases from Plasmodium Parasites : from Heterologous Production towards Functional Characterization". Thesis, Université Paris-Saclay (ComUE), 2018. http://www.theses.fr/2018SACLS447/document.
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Le paludisme est une maladie dévastatrice causée par un parasite du genre Plasmodium. Du fait de la propagation de souches résistantes aux actuels antipaludéens, il est nécessaire de comprendre les fonctions physiologiques essentielles du parasite afin de trouver de nouvelles cibles thérapeutiques. Les transporteurs membranaires sont une classe importante de cibles chez l'homme du fait de leur rôle physiologique essentiel pour la cellule. Cependant, chez les parasite du genre Plasmodium, seulement quelques transporteurs ont été biochimiquement caractérisés. Des études récentes de délétion de gènes dans un model murin ont montrées que l’ATPase de type P4, ou flippase, ATP2 de Plasmodium est essentielle pour le parasite. Chez les Eucaryotes, l’activité de translocation des lipides des ATPases de type P4 est nécessaire pour maintenir l’asymétrie des membranes, un élément clé dans de nombreux processus essentiels comme la formation de vésicules ou l’apoptose. Les flippases forment des complexes hétéromériques avec les protéines de la famille Cdc50 qui sont également trouvées dans le génome de Plasmodium. Pour comprendre le rôle fonctionnel de ces transporteurs putatifs durant l’infection par le parasite, nous avons besoin d’étudier leur mécanisme de transport et d’identifier leur (s) substrat (s). Nous avons entrepris l’expression hétérologue chez Saccharomyces cerevisiae d’ATP2, en complexe avec les sous unités Cdc50, de trois espèces différentes de Plasmodium. Nous avons réussi à co-exprimer l’orthologue ATP2 de P. chabaudi (PcATP2) et les sous unités PcCdc50 correspondantes. Par co-immunoprécipitation et une chromatographie d’exclusion stérique détectée par fluorescence, nous sommes parvenus à identifier la sous unité s’associant à PcATP2 : PcCdc50.1. Nous avons ensuite purifié le complexe PcATP2/PcCdc50.1 en utilisant des nanobodies reconnaissant la GFP fusionnée à l’extrémité C-terminale de PcATP2 et nous avons initié la caractérisation fonctionnelle avec des tests de phosphorylation et d’activité ATPasiqueMalaria is a devastating disease caused by a parasite of the genus Plasmodium. Due to the spread of strains resistant to current antimalarial drugs, it is necessary to understand essential physiological functions of the parasite in order to find new drug targets. Membrane transport proteins are an important class of drug targets in humans, as they perform essential physiological roles of the cell. However, for Plasmodium parasites, just a few membrane transporters have been biochemically described. Recent gene-deletion studies in malaria mouse models have shown that the Plasmodium P4-ATPase, or lipid flippase, ATP2 is essential for the parasite. In eukaryotes, the phospholipid translocation activity of P4-ATPases is needed to maintain the asymmetric distribution of membranes, a key element in many essential processes like vesicle budding or apoptosis. Lipid flippases form heteromeric complexes with members of the Cdc50 protein family, also found in the genomes of Plasmodium parasites. To understand the functional role of these still putative transporters during malaria infection we need to study their transport mechanism and identify their substrate(s). We have conducted the heterologous expression in Saccharomyces cerevisiae of ATP2 in complex with the Cdc50 subunits from three different Plasmodium species. We succeeded to co-express the ATP2 ortholog of P. chabaudi (PcATP2) and the related putative PcCdc50 proteins. By co-immunoprecipitation and Fluorescence-detection Size Exclusion Chromatography, we have managed to identify the Cdc50 β-subunit that associates to PcATP2: PcCdc50.1. We then purified the complex PcATP2/PcCdc50.1 using immobilized nanobodies that recognize the GFP fused at the C-terminal end of PcATP2 and we initiated the functional characterization using ATPase and phosphorylation activity assays
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Jacquot, Aurore. "Co-expression et caractérisation fonctionnelle d’un transporteur de lipides (une « flippase ») de la levure S. cerevisiae : l’ATPase P4 Drs2p, en complexe avec sa sous-unité associée Cdc50p". Thesis, Paris 11, 2012. http://www.theses.fr/2012PA11T081/document.
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Les membranes plasmiques et les membranes du trans-Golgi des cellules eucaryotes présentent une asymétrie des lipides qui les composent, avec les aminophospholipides (APLs : phosphatidylsérine et phosphatidyléthanolamine) enrichis dans le feuillet cytosolique. La dissipation de cette asymétrie est impliquée dans de nombreux processus (patho)physiologiques. Plusieurs études suggèrent que les ATPases P4 sont les candidats les plus probables pour le transport des APLs et le maintien de leur distribution asymétrique ; leur délétion dans la levure inhibe le trafic membranaire. En outre, des études ont montré que les ATPases P4 interagissaient avec les protéines de la famille CDC50 ; cette interaction est essentielle pour l’adressage et peut-être aussi la fonction des ATPases P4. Afin de contribuer à la compréhension du mécanisme de transport des lipides par les ATPases P4, l’objectif de ce travail a été de mettre au point la co-expression fonctionnelle, dans la levure, de l’ATPase P4 Drs2p et de sa protéine partenaire Cdc50p. Nous avons obtenu une fraction membranaire enrichie à 3% avec la protéine Drs2p, majoritairement en complexe avec Cdc50p. L’étude fonctionnelle du complexe nous a permis de mettre en évidence un rôle crucial du phosphatidylinositol-4-phosphate (PI(4)P), un important régulateur du trafic membranaire, au cours d’une étape particulière du cycle catalytique. Nous avons également développé un protocole de purification sur résine streptavidine du complexe Drs2p/Cdc50p. Enfin, comme un site potentiel d’interaction avec le PI(4)P est présent sur l’extrémité C-terminale de Drs2p, nous avons engendré différentes constructions de Drs2p, dans lesquelles une partie de l’extrémité C-terminale a été délétée ; dans une autre construction, l’extrémité N-terminale a également été délétée. Notre travail ouvre la voie à la caractérisation fonctionnelle et structurale détaillée du complexe Drs2p/Cdc50p, et à l’étude du rôle du transport de lipides dans le trafic membranaireTrans-Golgi membranes and plasma membranes of eukaryotic cells are asymmetric, with their cytosolic leaflet enriched in aminophospholipids (APLs: phosphatidylserine and phosphatidylethanolamine). Dissipation of this asymmetry is involved in many (patho)physiological processes. P4 ATPases are prime candidates for APL transport and for maintaining asymmetry across membranes. In addition, yeast deleted for P4 ATPases display membrane trafficking defects. Besides, CDC50 proteins have been shown to interact physically with P4 type ATPases, and this interaction is important for addressing the complex to the right destination, and possibly also for its function. To gain insight into the molecular mechanism of lipid transport by P4 ATPases, the goal of my thesis was to develop the co-expression, in yeast, of a functional P4 ATPase, Drs2p, together with its partner, Cdc50p. The strategy we developed allowed us to obtain a membrane fraction enriched in Drs2p (~3%), mainly in complex with Cdc50p. Functional characterization of the complex identified phosphatidylinositol-4-phosphate (PI4P), a major regulator of membrane trafficking, as a crucial component for rapid completion of the Drs2p/Cdc50p catalytic cycle. We also purified the complex in one step on streptavidin beads. Finally, we started investigating the potential auto-inhibitory roles of the C-terminus (as the C-terminus of Drs2p contains a PI4P binding site) and the N-terminus of Drs2p, by expressing various truncated versions of Drs2p. Our work sets the stage for detailed functional and structural characterization of the Drs2p/Cdc50p complex and its role in membrane traffic
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Dieudonne, Thibaud. "Functional and Structural Characterization of Lipid Flippases : The Yeast Drs2p/Cdc50p and the Disease-Related Human Atp8b1/Cdc50a Complexes Structure and Autoregulation of a P4-ATPase Lipid Flippase Screening of Detergents for Stabilization of Functional Membrane Proteins High phosphatidylinositol 4-phosphate (PI4P)-dependent ATPase activity for the Drs2p-Cdc50p flippase after removal of its N- and C-terminal extensions Slow Phospholipid Exchange between a Detergent-Solubilized Membrane Protein and Lipid-Detergent Mixed Micelles: Brominated Phospholipids as Tools to Follow Its Kinetics". Thesis, université Paris-Saclay, 2020. http://www.theses.fr/2020UPASS023.
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Les cellules sont entourées de membranes lipidiques organisées en bicouche séparant ainsi le milieu intracellulaire du milieu extérieur. L’une des caractéristiques des cellules eucaryotes est de posséder une distribution asymétrique des lipides constituants les membranes de la voie sécrétoire. En effet, dans ces membranes, la phosphatidylcholine (PC) et les sphingolipides (SL) sont majoritairement retrouvés sur le feuillet externe alors que la phosphatidylsérine (PS) et la phosphatidyléthanolamine (PE) sont séquestrées sur le feuillet interne. Cette asymétrie est maintenue grâce à la présence de transporteurs de lipides. Parmi ces transporteurs, on retrouve les flippases, qui grâce à l’énergie apportée par la consommation d’ATP, transportent les lipides du feuillet interne vers le feuillet externe. Les flippases appartiennent à la superfamille des ATPases de type P et ont été reliées à différentes pathologies humaines lorsqu’elles sont mutées. Par exemple, des mutations du gène ATP8B1 sont responsables d’une forme de cholestase intrahépatique, une maladie hépatique sévère. Dans cette thèse, nous avons étudier le mécanisme de régulation de deux flippases : la flippase de levure PS spécifique Drs2p/Cdc50p ainsi que la flippase humaine ATP8B1/CDC50A. Les deux flippases ont été exprimées dans la levure de bière S. cerevisiae et purifiées afin de réaliser leur caractérisation fonctionnelle. Nos résultats montrent que les deux flippases sont régulées par des phosphoinositides et auto-inhibées par leurs extrémités N- et C-terminalesLiving cells are surrounded by membranes organized in bilayers, separating the intracellular medium from the extracellular environment. A hallmark of eukaryotic membranes from the late secretory/endocytic pathways is the asymmetric distribution of phospholipids between the two leaflets. Indeed, phosphatidylcholine (PC) and sphingolipids (SL) are mainly found in the outer leaflet whereas phosphatidylserine (PS) and phosphatidylethanolamine (PE) are sequestered in the inner leaflet. This asymmetry is maintained thanks to different membrane lipid transporters. Among them, flippases, which are transporters fueled by ATP hydrolysis, translocate lipids from the outer to the inner leaflet. Flippases belong to the P4-ATPase family and have been linked to several diseases. For instance, mutated forms of a human P4-ATPase, ATP8B1, are responsible for intrahepatic cholestasis, a severe liver disease. In this thesis, we investigated the regulatory mechanism of two flippases, the yeast PS-specific flippase complex Drs2p/Cdc50p, and the human disease-related flippase complex ATP8B1/CDC50A. Both proteins were expressed in S. cerevisiae and purified for downstream functional characterization. Our results demonstrate that both flippases are tightly regulated by phosphoinositides and autoinhibited by their N- and C-terminal extensions
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Neutzner, Melanie. "Regulatoren des Zellteilungszyklus der Hefe Saccharomyces cerevisiae : die Polo-Kinase Cdc5 und der Ubiquitinierungsfaktor Hct1 /". [S.l. : s.n.], 2003. http://www.bsz-bw.de/cgi-bin/xvms.cgi?SWB10605153.
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Bizzari, Farid Fouad Mahmoud. "Cdc55 controls the balance of phosphatases to coordinate spindle assembly and chromosome disjunction during budding yeast meiosis". Thesis, University of Edinburgh, 2012. http://hdl.handle.net/1842/5876.
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Meiosis is the process by which haploid gametes are produced from a diploid cell. It is a specialised form of cell division which involves one round of DNA replication followed by two rounds of chromosome segregation. Errors in the segregation process can give rise to aneuploidy, which can result in miscarriages and birth defects. In the first meiotic division homologous chromosomes are segregated, and sister chromatids are segregated in the second division. This is coordinated with two rounds of spindle microtubule assembly and disassembly. How these two processes are coordinated is unknown. In my PhD, I study the role of the protein phosphatase 2A (PP2A) regulatory subunit, Cdc55, in budding yeast meiosis. PP2A is a conserved heterotrimeric enzyme that has important roles in mitosis and meiosis. These roles are dictated by binding to either of its two regulatory subunits, Rts1 and Cdc55, in budding yeast . I show that Cdc55 is required for the proper assembly of a meiotic spindle in meiosis I, through the maintenance of the Cdc14 phosphatase in the nucleolus early in meiosis. In addition, Cdc55 is also required to limit the formation of PP2A complexes with the Rts1 regulatory subunit, and this is essential for the timely dissolution of sister chromatid cohesion. Thus, Cdc55 couples spindle assembly with chromosome segregation through its interactions with Cdc14 and PP2ARts1. Finally, I show some preliminary studies looking at the possible downstream effectors of Cdc14 that are important in this mechanism.
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Coleman, Jonathan Allan. "P4-ATPase structure-function relationships : mechanism and roles of ATP8A2-CDC50A in aminophospholipid transport, protein trafficking, and visual disorders". Thesis, University of British Columbia, 2013. http://hdl.handle.net/2429/44073.
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P₄-ATPases are a family of membrane transporters which have been implicated in the energy-dependent transport of aminophospholipids from the exocytoplasmic to cytoplasmic surface of biological membranes. This thesis investigation examined the structure-function relationships of ATP8A2, a novel member of the P₄-ATPase family initially discovered in a proteomic study of photoreceptor outer segments. Photoreceptor outer segments are organelles which consist of stacks of membraneous discs containing visual pigment
molecules. ATP8A2 is shown to be present in photoreceptor outer segment discs and
preferentially transports phosphatidylserine towards the cytosolic leaflet, providing the first
direct demonstration of lipid transport by a purified mammalian P₄-ATPase. CDC50A, the β-subunit of ATP8A2 was discovered using mass spectrometry and Western blotting. Subunit interactions are mediated through the extracellular and membrane domains of CDC50A. The N-terminal domain of CDC50A appears to play a role in pump modulation. ATP8A2 forms a phosphoenzyme intermediate at Asp⁴¹⁶ and phosphatidylserine
appears to be transported in a similar manner to that of other cation-transporting P-type
ATPases. The phosphoenzyme exists in two distinct conformations: E₁P and E₂P. The E₂P form interacts with aminophospholipids. Lys⁸⁷³ in transmembrane segment M5 located in a region known to be important for cation binding for P-type ATPases is critical for phosphatidylserine binding. Glu¹⁹⁸ in the DGET motif is essential for E₂P dephosphorylation. ATP8A2 gene expression was disrupted in mice using a neo cassette. Knockout mice develop short outer segments and visual function is impaired. Modification of transbilayer
asymmetry and composition appear to be responsible for reduced visual function rather than structural defects. The decrease in outer segment length suggests that ATP8A2 is involved in vesicular trafficking of proteins to the outer segment by regulating the step of vesicle budding possibly from the trans-Golgi network. We speculate that ATP8A2 plays a similar role in other neuronal cells and thus provide insight into the phenotype of human disorders caused by mutations in ATP8A2.
In summary, this study has identified for the first time the transported substrate of a mammalian P₄-ATPase, discovered protein-protein interactions regulating function, elucidated the mechanism of lipid transport, and illuminated the function of ATP8A2 in photoreceptor and neuronal biology.
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Hysenaj, Lisiena. "Alterations of hematopoiesis during brucellosis". Thesis, Aix-Marseille, 2019. http://www.theses.fr/2019AIXM0251.
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La brucellose est une maladie qui se transmet de l’animal à l’homme. Elle est causée par la bactérie Brucella. Lors de ma thèse, j’ai montré que Brucella persiste dans les cellules de la moelle osseuse des animaux infectés. Ces observations sont très importantes car la moelle est un organe responsable de la génération des cellules du système immunitaires et c’est la principale niche des cellules souches hématopoïétiques. Au cours de ma thèse, j'ai montré que la protéine de la membrane externe 25 de Brucella (Omp25) est capable de lier au récepteur SLAMF1, une molécule exprimée par les cellules souches hématopoïétiques. Cette interaction conduit à la génération d'un plus grand nombre de cellules myéloïdes par les cellules souches hématopoïétiques. Les cellules myéloïdes sont la niche préférée de Brucella. Ainsi, cette stratégie permet à la bactérie d'envahir l’hôte et d'établir une infection chronique de longue durée. SLAMF 1 apparaît comme une nouvelle cible thérapeutique pour le contrôle des maladies infectieuses chroniques, ce qui représenterait une avancée importante dans la génération de nouveaux médicamentsBrucellosis is a disease that is transmitted from animals to humans. It is caused by the pathogenic bacterium Brucella. During my thesis, I showed that Brucella persists in the bone marrow cells of infected animals. These observations are very important because the bone marrow is an organ of the immune system responsible for the generation of the immune cells, as it is the principal niche of hematopoietic stem cells. During my thesis, I showed that Brucella outer membrane 25 (Omp25) is able to bind SLAMF1, a hematopoietic stem cell molecule. This interaction leads to the production of more myeloid cells by the hematopoietic stem cell. Myeloid cells are the favorite niche of Brucella. Thus, this strategy allows the bacteria to invade the host and establish a long lasting chronic infection. SLAMF 1 appears as a new therapeutic target for controlling chronic infectious diseases, which would represent an important advance in the generation of new drugs
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Genga, Kelly Roveran. "Estudo da imunoexpressão de proteínas relacionadas ao ponto de checagem mitótico (CDC20 e MAD2) e ao fuso mitótico (Aurora A e Aurora B) em pacientes portadores de síndrome mielodisplásica". reponame:Repositório Institucional da UFC, 2014. http://www.repositorio.ufc.br/handle/riufc/10358.
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GENGA, Kelly Roveran. Estudo da imunoexpressão de proteínas relacionadas ao ponto de checagem mitótico (CDC20 e MAD2) e ao fuso mitótico (Aurora A e Aurora B) em pacientes portadores de síndrome mielodisplásica. 2014. 145 f. Dissertação (Mestrado em Patologia) - Universidade Federal do Ceará. Faculdade de Medicina, Fortaleza, 2014.Submitted by denise santos (denise.santos@ufc.br) on 2014-12-22T13:55:37Z No. of bitstreams: 1 2014_dis_krgenga.pdf: 2677752 bytes, checksum: a4b47020e1d8a4b4789af7e144a1ce58 (MD5)
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Síndrome mielodisplásica (SMD) representa um grupo de doenças hematopoéticas heterogêneas, que se caracterizam por alterações morfológicas de dispoese, medula óssea hiperproliferativa, citopenias no sangue periférico e risco aumentado de evolução para leucemia mieloide aguda. A patogênese da SMD envolve múltiplas etapas, com destaque para alterações citogenéticas, porém ainda não está claramente definido como a doença progride. Nesse contexto, o estudo da imunoexpressão de proteínas relacionadas ao ponto de checagem mitótico (CDC20 e MAD2) e ao fuso mitótico (AURORA A e AURORA B) mostra-se promissor. Essas proteínas foram associadas à instabilidade cromossômica, aneuploidia e progressão tumoral em tumores sólidos e neoplasias hematológicas, contribuindo para o surgimento de anormalidades citogenéticas. Na literatura, existem poucos artigos publicados avaliando o impacto da expressão dessas proteínas na evolução de pacientes com SMD. O presente estudo, de caráter retrospectivo, teve como objetivo avaliar a imunoexpressão de CDC20, MAD2, AURORA A e AURORA B em pacientes com SMD e investigar sua relação com variáveis clínicas e laboratoriais. O estudo da expressão proteica foi realizado por imunoistoquímica (microscopia óptica), em 40 biópsias de medula óssea de pacientes portadores de SMD e 10 controles (biópsias de medula óssea para estadiamento de linfoma sem infiltração pela doença). A expressão proteica foi interpretada de forma qualitativa (expressão positiva versus negativa) e quantitativa (% de células positivas após análise de 10 campos em aumento de 400x, assim como pela categorização das amostras positivas em grupos de expressão leve - < 10%, moderada – 10 a 49% e alta - ≥ 50%). Identificou-se, de forma significativa, maior expressão das proteínas CDC20, MAD2 e AURORA B nos pacientes portadores de SMD, quando comparados com os controles (p < 0,05). A análise qualitativa mostrou os seguintes resultados (p < 0,05): 1) maior frequência de expressão negativa de MAD2 em pacientes com duas a três citopenias; 2) maior frequência de expressão negativa de CDC20 em pacientes com plaquetopenias mais graves (<50.000/mm3); 3) maior frequência de expressão positiva de AURORA B entre os pacientes que evoluíram para óbito; 4) menor sobrevida global entre os pacientes com expressão positiva de AURORA B; 5) maior frequência de expressão positiva de AURORA A entre os pacientes com dependência transfusional, citogenética aneuploide, cariótipo complexo e/ou anormal. A análise quantitativa (média ± DP, em %) mostrou os seguintes resultados (p < 0,05): 1) maior expressão de MAD2 e CDC20 em pacientes com plaquetopenias mais graves (<50.000/mm3); 2) maior expressão de MAD2 e CDC20 entre os pacientes que evoluíram para óbito; 3) maior expressão de CDC20 entre os pacientes com três displasias e cariótipo complexo; 4) maior expressão de AURORA B nos pacientes com citogenética alterada. A análise quantitativa por grupos revelou (p < 0,05): 1) maior frequência de alta expressão de CDC20 e MAD2 em pacientes com contagens plaquetárias < 50.000/mm3 e entre os pacientes que evoluíram para óbito durante o período do estudo; 2) menor sobrevida global no grupo de pacientes com expressão ≥ 50% de MAD2 e CDC20 e ≥ 10% de AURORA B. em comparação aos pacientes com expressão negativa e/ou expressão positiva < 50% e < 10%, respectivamente. Tais dados sugerem que expressões alteradas dessas proteínas (hipo ou hiperexpresssão) podem estar relacionadas ao processo de tumorigênese e à progressão da SMD, contribuindo, desse modo, para melhor estratificação de risco e abordagem terapêutica.
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Azouaoui, Hassina. "Étude structurale et fonctionnelle d’un transporteur de lipides « une flippase » de la levure S. cerevisiae : l’ATPase P4 Drs2p et sa sous unité-associée Cdc50p". Thesis, Université Paris-Saclay (ComUE), 2016. http://www.theses.fr/2016SACLS224/document.
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Les ATPases-P4 sont des transporteurs membranaires couplant l'hydrolyse de l'ATP au transport de lipides dans les membranes cellulaires eucaryotes. Avec leurs partenaires, les protéines CDC50, les ATPases-P4 transportent les phospholipides, en particulier la phosphatidylsérine (PS) et la phosphatidyléthanolamine (PE), du feuillet exoplasmique au feuillet cytosolique des membranes, assurant ainsi le maintien de l'asymétrie membranaire.Drs2p est l'une des cinq ATPases-P4 de la levure Saccharomyces cerevisiae. Elle est localisée dans les membranes du trans-Golgi (TGN), et elle a comme partenaire la protéine Cdc50p, qui est nécessaire à l'adressage correct et probablement au transport catalysé par Drs2p. Drs2p est principalement responsable du transport de la phosphatidylsérine (PS) dans les membranes du TGN et son activité est essentielle pour le maintien de la PS dans le feuillet cytosolique de ces membranes. En raison du rôle crucial de la PS dans de nombreuses voies de signalisation, aussi bien à l’extérieur (au cours de l’apoptose par exemple) qu’à l’intérieur de la cellule (par le recrutement de protéines impliquées dans des processus cellulaires essentiels), il est important de comprendre le mécanisme par lequel l’asymétrie de la PS est établie.Afin de progresser dans la compréhension du mécanisme moléculaire du transport de lipides, nous avons mis au point une procédure qui nous a permis de co-exprimer Drs2p et Cdc50p dans Saccharomyces cerevisiae. La purification de Drs2p par chromatographie d'affinité sur résine streptavidine a permis d'obtenir une fraction purifiée contenant très majoritairement Drs2p et Cdc50p, à raison de 1-2 mg/L de culture. Les deux protéines sont sous forme de complexe avec une stœchiométrie d'association de 1:1. Le complexe purifié est fonctionnel, et présente une activité d’hydrolyse de l’ATP stimulée par son substrat, la PS. Cette stimulation n’est cependant possible qu’en présence de PI4P, un phosphoinositide impliqué dans la régulation du trafic membranaire.De par leur rôle crucial dans le maintien de l'asymétrie membranaire, les ATPases-P4 ne peuvent qu'être régulées. Comme de nombreuses ATPases de type P sont soumises à une auto-régulation de leur activité, nous avons examiné la possibilité d’une telle auto-régulation dans le cas des ATPases P4. Pour ce faire, une approche par mutagenèse dirigée et protéolyse ménagée associée à l’identification par spectrométrie de masse des peptides ont été effectuées. La protéolyse ménagée du complexe purifié Drs2p/Cdc50p montre une activité ATPasique dépendante au PI4P de 30-50 fois plus importante. La protéolyse par la thrombine engendre un Drs2p dépourvu d'une partie N-terminale (R104) et d'une partie C-terminale (R1290) qui reste toujours associé à Cdc50p. Ce résultat montre qu'une coupure appropriée au niveau des extrémités terminales de Drs2p peut augmenter de façon significative, en présence du PI4P, l'activité ATPasique du complexe, nous amenant ainsi à identifier un rôle auto-inhibiteur des extrémités N- et/ou C-terminales de Drs2p.Ce travail ouvre des perspectives quant à la caractérisation structurale et fonctionnelle du mécanisme de transport de lipides par le complexe. Par ailleurs, il laisse entrevoir la possibilité d’étudier les bases moléculaires des pathologies associées aux mutations de certaines ATPases P4 humainesMaintenance of phospholipid asymmetry in eukaryotic cell membranes is essential for cellular integrity and function. P4-ATPases, from the P-type ATPases family, are energy-dependent transporters, together with their CDC50 accessory subunits couple ATP hydrolysis to lipid transport from the exoplasmic to cytoplasmic leaflet to maintain membrane asymmetry.Drs2p is one of these P4-ATPases in the yeast Saccharomyces cerevisiae. Drs2p is localised in trans-Golgi (TGN) membranes in association with its binding partner Cdc50p, which contributes to the correct addressing of Drs2p and probably in the catalyzed transport by Drs2p. Drs2p transport principally phosphatidylserine (PS) in TGN membranes. The PS is important for a several signalling pathways, for example, in apoptosis and recruitment of the proteins implied in various essential cellular process, so, it's very important to understand the mechanism that establishes this asymmetry.To gain in comprehension of molecular mechanism of lipid transport, robust protocols for expression and purification are required. In this work, we present a procedure for high-yield co-expression of Drs2p and Cdc50p. The purification of Drs2p and Cdc50p is achieved in a single step by affinity chromatography on streptavidin beads, yielding, 1-2 mg purified Drs2p/Cdc50p per liter of culture. This procedure allows purification of the complex Drs2p/Cdc50p with stoichiometry to 1:1. Our complex is functional, overal ATP hydrolysis by the complex is dependent of PS, favourite substrate of Drs2p. This hydrolyze is critically dependent on the presence of PI4P, a phosphoinositide involved in regulation of membrane trafficking.Like many P-type ATPases auto-regulate their activity, we examined the possibility that P4-ATPases are auto-regulated. In this work, we use directed mutagenesis and limited proteolysis associated with mass spectrometry for identify peptides. We show that limited proteolysis of a purified complex Drs2p/Cdc50p resulted in up to a 30-50 fold increase of it ATPase activity, which however remained dependent on PI4P. Using thrombin as the protease, Cdc50p remained intact and in complex with Drs2p, which was cleaved at two positions, namely after R104 and after R1290. Our results therefore reveal that trimming off appropriate regions of the terminal extensions of Drs2p can increase its ATPase activity in the presence of PI4P by an enormous factor, thereby identifying a role of N and/or C-terminal extensions in auto-inhibition of Drs2p.Our results open perspectives on the structural and the functional characterization of the lipid transport mechanism by the complex Drs2p/Cdc50p. Furthermore, our procedures open up the possibility of studying the molecular bases of the pathologies associated with the mutations of human P4-ATPases
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Vendrell, Arasa Alexandre. "SCF cdc4 regulates msn2 and msn4 dependent gene expression to counteract hog1 induced lethality". Doctoral thesis, Universitat Pompeu Fabra, 2009. http://hdl.handle.net/10803/7153.
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L'activació sostinguda de Hog1 porta a una inhibició del creixement cel·lular. En aquest treball, hem observat que el fenotip de letalitat causat per l'activació sostinguda de Hog1 és parcialment inhibida per la mutació del complexe SCFCDC4. La inhibició de la mort causada per l'activació sostinguda de Hog1 depèn de la via d'extensió de la vida. Quan Hog1 s'activa de manera sostinguda, la mutació al complexe SCFCDC4 fa que augmenti l'expressió gènica depenent de Msn2 i Msn4 que condueix a una sobreexpressió del gen PNC1 i a una hiperactivació de la deacetilassa Sir2. La hiperactivació de Sir2 és capaç d'inhibir la mort causada per l'activació sostinguda de Hog1.També hem observat que la mort cel·lular causada per l'activació sostinguda de Hog1 és deguda a una inducció d'apoptosi. L'apoptosi induïda per Hog1 és inhibida per la mutació al complexe SCFCDC4. Per tant, la via d'extensió de la vida és capaç de prevenir l'apoptosi a través d'un mecanisme desconegut.
Sustained Hog1 activation leads to an inhibition of cell growth. In this work, we have observed that the lethal phenotype caused by sustained Hog1 activation is prevented by SCFCDC4 mutants. The prevention of Hog1-induced cell death by SCFCDC4 mutation depends on the lifespan extension pathway. Upon sustained Hog1 activation, SCFCDC4 mutation increases Msn2 and Msn4 dependent gene expression that leads to a PNC1 overexpression and a Sir2 deacetylase hyperactivation. Then, hyperactivation of Sir2 is able to prevent cell death caused by sustained Hog1 activation.
We have also observed that cell death upon sustained Hog1 activation is due to an induction of apoptosis. The apoptosis induced by Hog1 is decreased by SCFCDC4 mutation. Therefore, lifespan extension pathway is able to prevent apoptosis by an unknown mechanism.
Capítulos de libros sobre el tema "CDC50 proteins"
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Montigny, Cédric, Hassina Azouaoui, Aurore Jacquot, Marc le Maire, Christine Jaxel, Philippe Champeil y Guillaume Lenoir. "Overexpression of Membrane Proteins in Saccharomyces cerevisiae for Structural and Functional Studies: A Focus on the Rabbit Ca2+-ATPase Serca1a and on the Yeast Lipid “Flippase” Complex Drs2p/Cdc50p". En Membrane Proteins Production for Structural Analysis, 133–71. New York, NY: Springer New York, 2014. http://dx.doi.org/10.1007/978-1-4939-0662-8_6.
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"SCF (Skp1—CDC53—F-box protein)". En Encyclopedia of Genetics, Genomics, Proteomics and Informatics, 1762. Dordrecht: Springer Netherlands, 2008. http://dx.doi.org/10.1007/978-1-4020-6754-9_15111.
Texto completoActas de conferencias sobre el tema "CDC50 proteins"
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Nath, Somsubhra, Taraswi Banerjee, Debrup Sen, Tania Das y Susanta Roychoudhury. "Abstract 3075: A novel transcriptional role of spindle assembly checkpoint protein Cdc20 regulating the expression of mitotic ubiquitin carrier protein UbcH10". En Proceedings: AACR 102nd Annual Meeting 2011‐‐ Apr 2‐6, 2011; Orlando, FL. American Association for Cancer Research, 2011. http://dx.doi.org/10.1158/1538-7445.am2011-3075.
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Hu, Die, Brigitte L. Thériault, Vida Talebian, Julie Owen, Richard Marcellus y Rima Al-awar. "Abstract 1222: Validating the WD repeat protein CDC40 as a potential therapeutic target for lung cancer". En Proceedings: AACR Annual Meeting 2021; April 10-15, 2021 and May 17-21, 2021; Philadelphia, PA. American Association for Cancer Research, 2021. http://dx.doi.org/10.1158/1538-7445.am2021-1222.
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