Structural basis for activation, assembly and membrane binding of ESCRT-III Snf7 filaments

Shaogeng Tang, W. Mike Henne, Peter P. Borbat, Nicholas J. Buchkovich, Jack H. Freed, Yuxin Mao, J. Christopher Fromme, Scott D. Emr

Research output: Contribution to journalArticle

51 Scopus citations

Abstract

The endosomal sorting complexes required for transport (ESCRTs) constitute hetero- oligomeric machines that catalyze multiple topologically similar membrane-remodeling processes. Although ESCRT-III subunits polymerize into spirals, how individual ESCRT-III subunits are activated and assembled together into a membrane-deforming filament remains unknown. Here, we determine X-ray crystal structures of the most abundant ESCRT-III subunit Snf7 in its active conformation. Using pulsed dipolar electron spin resonance spectroscopy (PDS), we show that Snf7 activation requires a prominent conformational rearrangement to expose protein-membrane and protein-protein interfaces. This promotes the assembly of Snf7 arrays with ~30 Å periodicity into a membrane-sculpting filament. Using a combination of biochemical and genetic approaches, both in vitro and in vivo, we demonstrate that mutations on these protein interfaces halt Snf7 assembly and block ESCRT function. The architecture of the activated and membrane-bound Snf7 polymer provides crucial insights into the spatially unique ESCRT-III-mediated membrane remodeling.

Original languageEnglish (US)
Article numbere12548
JournaleLife
Volume4
Issue numberDECEMBER2015
DOIs
StatePublished - Dec 15 2015

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)
  • Immunology and Microbiology(all)
  • Biochemistry, Genetics and Molecular Biology(all)

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    Tang, S., Henne, W. M., Borbat, P. P., Buchkovich, N. J., Freed, J. H., Mao, Y., Fromme, J. C., & Emr, S. D. (2015). Structural basis for activation, assembly and membrane binding of ESCRT-III Snf7 filaments. eLife, 4(DECEMBER2015), [e12548]. https://doi.org/10.7554/eLife.12548