Cep55, a microtubule-bundling protein, associates with centralspindlin to control the midbody integrity and cell abscission during cytokinesis.
We report here an efficient functional genomic analysis by combining information on the gene expression profiling, cellular localization, and loss-of-function studies. Through this analysis, we identified Cep55 as a regulator required for the completion of cytokinesis. We found that Cep55 localizes to the mitotic spindle during prometaphase and metaphase and to the spindle midzone and the midbody during anaphase and cytokinesis. At the terminal stage of cytokinesis, Cep55 is required for the midbody structure and for the completion of cytokinesis. In Cep55-knockdown cells, the Flemming body is absent, and the structural and regulatory components of the midbody are either absent or mislocalized. Cep55 also facilitates the membrane fusion at the terminal stage of cytokinesis by controlling the localization of endobrevin, a v-SNARE required for cell abscission. Biochemically, Cep55 is a microtubule-associated protein that efficiently bundles microtubules. Cep55 directly binds to MKLP1 in vitro and associates with the MKLP1-MgcRacGAP centralspindlin complex in vivo. Cep55 is under the control of centralspindlin, as knockdown of centralspindlin abolished the localization of Cep55 to the spindle midzone. Our study defines a cellular mechanism that links centralspindlin to Cep55, which, in turn, controls the midbody structure and membrane fusion at the terminal stage of cytokinesis.
Zhao
Wei-meng
WM
Department of Biological Sciences, Stanford University, Stanford, CA 94305-5020, USA.
Seki
Akiko
A
Fang
Guowei
G
D000705:Anaphase; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002478:Cells, Cultured; D048749:Cytokinesis; D020690:GTPase-Activating Proteins / Q000378:metabolism*; D015894:Genome, Human / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism*; D011485:Protein Binding; D021381:Protein Transport; D050683:R-SNARE Proteins / Q000378:metabolism; D008941:Spindle Apparatus / Q000378:metabolism*
D018797:Cell Cycle Proteins; C505378:Cep55 protein, human; D020690:GTPase-Activating Proteins; C092935:KIF23 protein, human; D008869:Microtubule-Associated Proteins; D009687:Nuclear Proteins; D050683:R-SNARE Proteins; C112938:VAMP8 protein, human; C115995:mgcRacGAP