TY - JOUR
T1 - Structural basis of CHMP2A–CHMP3 ESCRT-III polymer assembly and membrane cleavage
AU - Azad, Kimi
AU - Guilligay, Delphine
AU - Boscheron, Cecile
AU - Maity, Sourav
AU - De Franceschi, Nicola
AU - Sulbaran, Guidenn
AU - Effantin, Gregory
AU - Wang, Haiyan
AU - Kleman, Jean Philippe
AU - Bassereau, Patricia
AU - Schoehn, Guy
AU - Roos, Wouter H.
AU - Desfosses, Ambroise
AU - Weissenhorn, Winfried
N1 - Funding Information:
This research was funded by the ANR (grant nos. ANR-14-CE09-0003-01 and ANR-19-CE11-0002-02 to W.W.). W.W. acknowledges support from the Institut Universitaire de France and access to the platforms of the Grenoble Instruct-ERIC center (IBS and ISBG; grant no. UAR 3518 CNRS-CEA-UGA-EMBL) within the Grenoble Partnership for Structural Biology, with support from FRISBI (grant no. ANR-10-INBS-05-02) and GRAL, a project of the University Grenoble Alpes graduate school (Ecoles Universitaires de Recherche) CBH-EUR-GS (grant no. ANR-17-EURE-0003). The IBS electron microscope facility is supported by the Auvergne-Rhône-Alpes Region, the Fondation pour la Recherche Medicale, the FEDER/ERDF fund (European Regional Development Fund) and the GIS-IBiSA (Infrastructures en Biologie, Sante et Agronomie). We acknowledge the provision of in-house experimental time from the CM01 facility at the European Synchrotron Radiation Facility and we thank L. Estrozi for extensive discussion and help with helical image analysis. We further thank the HIV Reagent Program, Division of AIDS, NIAID, NIH for providing the Anti-Human Immunodeficiency Virus 1 (HIV-1) p24 Monoclonal (183-H12-5C), ARP-3537, contributed by B. Chesebro and K. Wehrly.
Publisher Copyright:
© 2023, The Author(s), under exclusive licence to Springer Nature America, Inc.
PY - 2023/1/5
Y1 - 2023/1/5
N2 - The endosomal sorting complex required for transport (ESCRT) is a highly conserved protein machinery that drives a divers set of physiological and pathological membrane remodeling processes. However, the structural basis of ESCRT-III polymers stabilizing, constricting and cleaving negatively curved membranes is yet unknown. Here we present cryo-EM structures of membrane-coated CHMP2A–CHMP3 filaments from Homo sapiens of two different diameters at 3.3 and 3.6 Å resolution. The structures reveal helical filaments assembled by CHMP2A–CHMP3 heterodimers in the open ESCRT-III conformation, which generates a partially positive charged membrane interaction surface, positions short N-terminal motifs for membrane interaction and the C-terminal VPS4 target sequence toward the tube interior. Inter-filament interactions are electrostatic, which may facilitate filament sliding upon VPS4-mediated polymer remodeling. Fluorescence microscopy as well as high-speed atomic force microscopy imaging corroborate that VPS4 can constrict and cleave CHMP2A–CHMP3 membrane tubes. We therefore conclude that CHMP2A–CHMP3–VPS4 act as a minimal membrane fission machinery.
AB - The endosomal sorting complex required for transport (ESCRT) is a highly conserved protein machinery that drives a divers set of physiological and pathological membrane remodeling processes. However, the structural basis of ESCRT-III polymers stabilizing, constricting and cleaving negatively curved membranes is yet unknown. Here we present cryo-EM structures of membrane-coated CHMP2A–CHMP3 filaments from Homo sapiens of two different diameters at 3.3 and 3.6 Å resolution. The structures reveal helical filaments assembled by CHMP2A–CHMP3 heterodimers in the open ESCRT-III conformation, which generates a partially positive charged membrane interaction surface, positions short N-terminal motifs for membrane interaction and the C-terminal VPS4 target sequence toward the tube interior. Inter-filament interactions are electrostatic, which may facilitate filament sliding upon VPS4-mediated polymer remodeling. Fluorescence microscopy as well as high-speed atomic force microscopy imaging corroborate that VPS4 can constrict and cleave CHMP2A–CHMP3 membrane tubes. We therefore conclude that CHMP2A–CHMP3–VPS4 act as a minimal membrane fission machinery.
UR - http://www.scopus.com/inward/record.url?scp=85145672554&partnerID=8YFLogxK
U2 - 10.1038/s41594-022-00867-8
DO - 10.1038/s41594-022-00867-8
M3 - Article
C2 - 36604498
AN - SCOPUS:85145672554
SN - 1545-9993
VL - 30
SP - 81
EP - 90
JO - Nature Structural and Molecular Biology
JF - Nature Structural and Molecular Biology
ER -