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      저자 : Duiping Feng (검색결과 1 건)
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        EBP50 Phosphorylation by Cdc2/Cyclin B Kinase Affects Actin Cytoskeleton Reorganization and Regulates Functions of Human Breast Cancer Cell Line MDA-MB-231

        Chaoyuan Sun,Junqi He,Junfang Zheng,Shan Cheng,Duiping Feng 한국분자세포생물학회 2013 Molecules and cells Vol.36 No.1

        The actin cytoskeleton plays an important role in cell shape determination, adhesion and cell cycle progression. Ezrin-radixin-moesin (ERM)-binding phosphoprotein 50 (EBP50), also known as Na+-H+ exchanger regulatory factor 1 (NHERF1), associates with actin cytoskeleton and is related to cell cycle progression. Its Ser279 and Ser301 residues are phosphorylated by cyclin-dependent kinase 2 (cdc2)/cyclin B during the mitosis phase. However, the biological significance of EBP50 phosphorylation mediated by cdc2/cyclin B is not clear. In the present study, MDA-MB-231 cells with low levels of endogenous EBP50 protein were stably trans-fected with constructs of EBP50 wild type (WT), phospho-deficient (serine 279 and serine 301 mutated to alanine-S279A/S301A) or phospho-mimetic (serine 279 and serine 301 mutated to aspartic acid-S279D/S301D) mutants. Subsequently, multiple phenotypes of these cells were characterized. Failure of cdc2/cyclin B-mediated EBP50 phosphorylation in cells expressing S279A/S301A (AA cells) significantly increased F-actin content, enhanced the adherence of cells to the extracellular matrix, altered cell morphology and caused defects in cytokinesis, as reflected in the formation of giant cells with heteroploid DNA and multinucleation or giant nuclei. Furthermore, knockdown of EBP50 expres-sion in AA cells rescued cell defects such as the cytokinesis failure and abnormal cell morphology. EBP50 S279A/ S301A had a weaker binding affinity with actin than EBP50 S279D/S301D, which might explain the increase of F-actin content in the AA cells. The present results suggest that cdc2/cyclin B-mediated EBP50 phos-phorylation may play a role in the regulation of various cell functions by affecting actin cytoskeleton reorgan-ization.

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