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Published August 1, 2013 | Supplemental Material + Published
Journal Article Open

Electron cryotomography of ESCRT assemblies and dividing Sulfolobus cells suggests that spiraling filaments are involved in membrane scission

Abstract

The endosomal-sorting complex required for transport (ESCRT) is evolutionarily conserved from Archaea to eukaryotes. The complex drives membrane scission events in a range of processes, including cytokinesis in Metazoa and some Archaea. CdvA is the protein in Archaea that recruits ESCRT-III to the membrane. Using electron cryotomography (ECT), we find that CdvA polymerizes into helical filaments wrapped around liposomes. ESCRT-III proteins are responsible for the cinching of membranes and have been shown to assemble into helical tubes in vitro, but here we show that they also can form nested tubes and nested cones, which reveal surprisingly numerous and versatile contacts. To observe the ESCRT–CdvA complex in a physiological context, we used ECT to image the archaeon Sulfolobus acidocaldarius and observed a distinct protein belt at the leading edge of constriction furrows in dividing cells. The known dimensions of ESCRT-III proteins constrain their possible orientations within each of these structures and point to the involvement of spiraling filaments in membrane scission.

Additional Information

© 2013 Dobro et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). Received: Nov 6, 2012; Revised: May 28, 2013; Accepted: May 31, 2013. This article was published online ahead of print in MBoC in Press (http://www .molbiolcell.org/cgi/doi/10.1091/mbc.E12-11-0785) on June 12, 2013. This work was supported in part by National Institutes of Heath Grant P50 GM082545 to G.J.J., National Science Foundation Grant DMR1105277 to P.L.C., and a gift to Caltech from the Gordon and Betty Moore Foundation. We thank Wesley I. Sundquist for advice and critical reading of the manuscript, Kay Grunewald for use of laboratory equipment, and Morgan Beeby, Jason Porath, and Jean Choi for their help with illustrations.

Attached Files

Published - Mol._Biol._Cell-2013-Dobro-2319-27.pdf

Supplemental Material - CombinedSupMats.pdf

Supplemental Material - mc-E12-11-0785-s01.mov

Supplemental Material - mc-E12-11-0785-s02.wmv

Supplemental Material - mc-E12-11-0785-s03.mov

Supplemental Material - mc-E12-11-0785-s04.mov

Supplemental Material - mc-E12-11-0785-s05.mov

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August 19, 2023
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