Abstract
The final cytokinesis event involves severing of the connecting intercellular bridge (ICB) between daughter cells. FIP3-positive recycling endosomes (FIP3 endosomes) and ESCRT complexes have been implicated in mediating the final stages of cytokinesis. Here we analyse the spatiotemporal dynamics of the actin cytoskeleton, FIP3-endosome fusion and ESCRT-III localization during cytokinesis to show that the ICB narrows by a FIP3-endosome-mediated secondary ingression, whereas the ESCRT-III complex is needed only for the last scission step of cytokinesis. We characterize the role of FIP3 endosomes during cytokinesis to demonstrate that FIP3 endosomes deliver SCAMP2/3 and p50RhoGAP to the ICB during late telophase, proteins required for the formation of the secondary ingression. We also show that the FIP3-endosome-induced secondary ingression is required for the recruitment of the ESCRT-III complex to the abscission site. Finally, we characterize a FIP3-endosome-dependent regulation of the ICB cortical actin network through the delivery of p50RhoGAP. These results provide a framework for the coordinated efforts of actin, FIP3 endosomes and the ESCRTs to regulate cytokinesis and abscission.
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Acknowledgements
We thank C. Pearson and C. Willenborg (University of Colorado Denver, USA) for critically reading the manuscript. We also thank J. Martin-Serrano (King’s College London, UK) for mCherry–CEP55, mCherry–TSG101 and YFP–CHMP4B; M. Geiszt (Semmelweis University, Hungary) for GFP–p50RhoGAP; C. Westlake (NCI-Frederick, USA) for RAB8 complementary DNA; A. Bresnick (Albert Einstein College of Medicine, USA) for myosin-IIA–GFP; and W. Bement (University of Wisconsin Madison, USA) for utrophin–GFP and utrophin–RFP. This work was supported in part by a grant from NIH-NIDDK (DK064380 to R.P.).
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J.A.S. and R.P. conceived the project and wrote the manuscript with the help of D.C. and C.C.W. Proteomic round one was performed and analysed by G.C.S. and J.W. Proteomic round two was performed and analysed by J.A.S. and J.W. Proteomic round three was performed and analysed by J.A.S. and C.Z. All remaining experiments and data analysis were performed by J.A.S. and R.P.
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Schiel, J., Simon, G., Zaharris, C. et al. FIP3-endosome-dependent formation of the secondary ingression mediates ESCRT-III recruitment during cytokinesis. Nat Cell Biol 14, 1068–1078 (2012). https://doi.org/10.1038/ncb2577
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DOI: https://doi.org/10.1038/ncb2577
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