REL 2020-07-27 2020-11-18 2020-11-18 0000-0003-3721-5104 Stefan T Huber Department of Bionanoscience / TU Delft Van der Maasweg 9 Delft Zuid-Holland Germany 52428 0000-0002-1168-5143 Carsten Sachse Forschungszentrum Jülich GmbH Wilhelm-Johnen-Straße Juelich NRW Germany 52425 Huber ST Mostafavi S Mortensen SA Sachse C 70.0 10.6019/EMPIAR-10495 EMDB EMD-11212 Huber ST Mostafavi S Mortensen SA Sachse C Science Advances Sci.Adv United States 34 6 2020 English 10.1126/sciadv.aba4897

ESCRT-III proteins mediate a range of cellular membrane remodeling activities such as multivesicular body biogenesis, cytokinesis, and viral release. Critical to these processes is the assembly of ESCRT-III subunits into polymeric structures. In this study, we determined the cryo-EM structure of a helical assembly of Saccharomyces cerevisiae Vps24 at 3.2-Å resolution and found that Vps24 adopts an elongated open conformation. Vps24 forms a domain-swapped dimer extended into protofilaments that associate into a double-stranded apolar filament. We demonstrate that, upon binding negatively charged lipids, Vps24 homopolymer filaments undergo partial disassembly into shorter filament fragments and oligomers. Upon the addition of Vps24, Vps2, and Snf7, liposomes are deformed into neck and tubular structures by an ESCRT-III heteropolymer coat. The filamentous Vps24 homopolymer assembly structure and interaction studies reveal how Vps24 could introduce unique geometric properties to mixed-type ESCRT-III heteropolymers and contribute to the process of membrane scission events.

Aligned micrographs of Vps24 helical filaments /data micrographs - single frame MRC MRC 1320 1 32 BIT FLOAT 3838 1.04 3710 1.04

Selection of 1320 K2 micrographs based on specimen coverage, ice quality, and minimal drift. The same micrographs were used in the paper. Movies were motion-corrected and dose-weighted using MotionCor2. 40 e-/A2 cumulative exposure, 0.75 um to 3 um defocus range. 4 images per hole were taken on Quantifoil R2/1 grids.

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