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

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

Research output: Contribution to journalArticle

37 Citations (Scopus)

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

Fingerprint

Endosomal Sorting Complexes Required for Transport
Chemical activation
Membranes
Electron spin resonance spectroscopy
Proteins
Electron Spin Resonance Spectroscopy
Periodicity
Conformations
Molecular Biology
Polymers
Membrane Proteins
Crystal structure
X-Rays
X rays
Mutation

All Science Journal Classification (ASJC) codes

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

Cite this

Tang, S., Henne, W. M., Borbat, P. P., Buchkovich, N., Freed, J. H., Mao, Y., ... 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
Tang, Shaogeng ; Henne, W. Mike ; Borbat, Peter P. ; Buchkovich, Nicholas ; Freed, Jack H. ; Mao, Yuxin ; Fromme, J. Christopher ; Emr, Scott D. / Structural basis for activation, assembly and membrane binding of ESCRT-III Snf7 filaments. In: eLife. 2015 ; Vol. 4, No. DECEMBER2015.
@article{59b661da2f6343969cd26c4b139f682c,
title = "Structural basis for activation, assembly and membrane binding of ESCRT-III Snf7 filaments",
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 {\AA} 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.",
author = "Shaogeng Tang and Henne, {W. Mike} and Borbat, {Peter P.} and Nicholas Buchkovich and Freed, {Jack H.} and Yuxin Mao and Fromme, {J. Christopher} and Emr, {Scott D.}",
year = "2015",
month = "12",
day = "15",
doi = "10.7554/eLife.12548",
language = "English (US)",
volume = "4",
journal = "eLife",
issn = "2050-084X",
publisher = "eLife Sciences Publications",
number = "DECEMBER2015",

}

Tang, S, Henne, WM, Borbat, PP, Buchkovich, N, Freed, JH, Mao, Y, Fromme, JC & Emr, SD 2015, 'Structural basis for activation, assembly and membrane binding of ESCRT-III Snf7 filaments', eLife, vol. 4, no. DECEMBER2015, e12548. https://doi.org/10.7554/eLife.12548

Structural basis for activation, assembly and membrane binding of ESCRT-III Snf7 filaments. / Tang, Shaogeng; Henne, W. Mike; Borbat, Peter P.; Buchkovich, Nicholas; Freed, Jack H.; Mao, Yuxin; Fromme, J. Christopher; Emr, Scott D.

In: eLife, Vol. 4, No. DECEMBER2015, e12548, 15.12.2015.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Structural basis for activation, assembly and membrane binding of ESCRT-III Snf7 filaments

AU - Tang, Shaogeng

AU - Henne, W. Mike

AU - Borbat, Peter P.

AU - Buchkovich, Nicholas

AU - Freed, Jack H.

AU - Mao, Yuxin

AU - Fromme, J. Christopher

AU - Emr, Scott D.

PY - 2015/12/15

Y1 - 2015/12/15

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84988603419&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84988603419&partnerID=8YFLogxK

U2 - 10.7554/eLife.12548

DO - 10.7554/eLife.12548

M3 - Article

C2 - 26670543

AN - SCOPUS:84988603419

VL - 4

JO - eLife

JF - eLife

SN - 2050-084X

IS - DECEMBER2015

M1 - e12548

ER -