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Structural analysis and modeling reveals new mechanisms governing ESCRT-III spiral filament assembly.


ABSTRACT: The scission of biological membranes is facilitated by a variety of protein complexes that bind and manipulate lipid bilayers. ESCRT-III (endosomal sorting complex required for transport III) filaments mediate membrane scission during the ostensibly disparate processes of multivesicular endosome biogenesis, cytokinesis, and retroviral budding. However, mechanisms by which ESCRT-III subunits assemble into a polymer remain unknown. Using cryogenic electron microscopy (cryo-EM), we found that the full-length ESCRT-III subunit Vps32/CHMP4B spontaneously forms single-stranded spiral filaments. The resolution afforded by two-dimensional cryo-EM combined with molecular dynamics simulations revealed that individual Vps32/CHMP4B monomers within a filament are flexible and able to accommodate a range of bending angles. In contrast, the interface between monomers is stable and refractory to changes in conformation. We additionally found that the carboxyl terminus of Vps32/CHMP4B plays a key role in restricting the lateral association of filaments. Our findings highlight new mechanisms by which ESCRT-III filaments assemble to generate a unique polymer capable of membrane remodeling in multiple cellular contexts.

SUBMITTER: Shen QT 

PROVIDER: S-EPMC4164947 | biostudies-literature | 2014 Sep

REPOSITORIES: biostudies-literature

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Structural analysis and modeling reveals new mechanisms governing ESCRT-III spiral filament assembly.

Shen Qing-Tao QT   Schuh Amber L AL   Zheng Yuqing Y   Quinney Kyle K   Wang Lei L   Hanna Michael M   Mitchell Julie C JC   Otegui Marisa S MS   Ahlquist Paul P   Cui Qiang Q   Audhya Anjon A  

The Journal of cell biology 20140908 6


The scission of biological membranes is facilitated by a variety of protein complexes that bind and manipulate lipid bilayers. ESCRT-III (endosomal sorting complex required for transport III) filaments mediate membrane scission during the ostensibly disparate processes of multivesicular endosome biogenesis, cytokinesis, and retroviral budding. However, mechanisms by which ESCRT-III subunits assemble into a polymer remain unknown. Using cryogenic electron microscopy (cryo-EM), we found that the f  ...[more]

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