Cortical Constriction During Abscission Involves Helices of ESCRT-III–Dependent Filaments

The process by which animal cells are physically separated after cell division is dissected in molecular detail. After partitioning of cytoplasmic contents by cleavage furrow ingression, animal cells remain connected by an intercellular bridge, which subsequently splits by abscission. Here, we examined intermediate stages of abscission in human cells by using live imaging, three-dimensional structured illumination microscopy, and electron tomography. We identified helices of 17-nanometer-diameter filaments, which narrowed the cortex of the intercellular bridge to a single stalk. The endosomal sorting complex required for transport (ESCRT)–III co-localized with constriction zones and was required for assembly of 17-nanometer-diameter filaments. Simultaneous spastin-mediated removal of underlying microtubules enabled full constriction at the abscission site. The identification of contractile filament helices at the intercellular bridge has broad implications for the understanding of cell division and of ESCRT-III–mediated fission of large membrane structures.

[1]  S. Emr,et al.  Structure and disassembly of filaments formed by the ESCRT-III subunit Vps24. , 2008, Structure.

[2]  J. Lippincott-Schwartz,et al.  Brefeldin A: insights into the control of membrane traffic and organelle structure , 1992, The Journal of cell biology.

[3]  J. Luzio,et al.  Spastin Couples Microtubule Severing to Membrane Traffic in Completion of Cytokinesis and Secretion , 2008, Traffic.

[4]  D. I. Svergun,et al.  A crescent-shaped ALIX dimer targets ESCRT-III CHMP4 filaments. , 2009, Structure.

[5]  J. Martin-Serrano,et al.  Differential requirements for Alix and ESCRT-III in cytokinesis and HIV-1 release , 2008, Proceedings of the National Academy of Sciences.

[6]  C. Mandato,et al.  Wound-induced contractile ring: a model for cytokinesis. , 2005, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[7]  J. Martin-Serrano,et al.  Parallels Between Cytokinesis and Retroviral Budding: A Role for the ESCRT Machinery , 2007, Science.

[8]  Natalie Elia,et al.  Midbody Targeting of the ESCRT Machinery by a Noncanonical Coiled Coil in CEP55 , 2008, Science.

[9]  J. Hurley,et al.  Membrane budding and scission by the ESCRT machinery: it's all in the neck , 2010, Nature Reviews Molecular Cell Biology.

[10]  D. L. Taylor,et al.  Traction forces of cytokinesis measured with optically modified elastic substrata , 1997, Nature.

[11]  M. Gustafsson,et al.  Subdiffraction Multicolor Imaging of the Nuclear Periphery with 3D Structured Illumination Microscopy , 2008, Science.

[12]  S. Emr,et al.  Functional Reconstitution of ESCRT-III Assembly and Disassembly , 2009, Cell.

[13]  I. Mellman,et al.  The Rab8 GTPase selectively regulates AP-1B–dependent basolateral transport in polarized Madin-Darby canine kidney cells , 2003, The Journal of cell biology.

[14]  D. Gerlich,et al.  Aurora B-Mediated Abscission Checkpoint Protects against Tetraploidization , 2009, Cell.

[15]  P. Hanson,et al.  Plasma membrane deformation by circular arrays of ESCRT-III protein filaments , 2008, The Journal of cell biology.

[16]  S. Gygi,et al.  Human ESCRT and ALIX proteins interact with proteins of the midbody and function in cytokinesis , 2007, The EMBO journal.

[17]  J. Jaén,et al.  Selective, covalent modification of beta-tubulin residue Cys-239 by T138067, an antitumor agent with in vivo efficacy against multidrug-resistant tumors. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[18]  Jacob Piehler,et al.  Helical Structures of ESCRT-III Are Disassembled by VPS4 , 2008, Science.

[19]  D. Gerlich,et al.  Cytokinetic abscission: cellular dynamics at the midbody. , 2009, Trends in cell biology.

[20]  F. Barr,et al.  Cytokinesis: Placing and Making the Final Cut , 2007, Cell.

[21]  Mullins Jm,et al.  Terminal phase of cytokinesis in D-98S cells , 1977 .

[22]  Timothy J Mitchison,et al.  Animal cytokinesis: from parts list to mechanisms. , 2006, Annual review of biochemistry.

[23]  S. Redick,et al.  Centriolin Anchoring of Exocyst and SNARE Complexes at the Midbody Is Required for Secretory-Vesicle-Mediated Abscission , 2005, Cell.

[24]  P. Chavrier,et al.  Endocytic traffic in animal cell cytokinesis. , 2008, Current opinion in cell biology.

[25]  J. Lippincott-Schwartz,et al.  Structural basis for midbody targeting of spastin by the ESCRT-III protein CHMP1B , 2008, Nature Structural &Molecular Biology.

[26]  Jennifer Lippincott-Schwartz,et al.  Membrane scission by the ESCRT-III complex , 2009, Nature.