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Elife ; 32014 May 30.
Article in English | MEDLINE | ID: mdl-24878737

ABSTRACT

The ESCRT machinery along with the AAA+ ATPase Vps4 drive membrane scission for trafficking into multivesicular bodies in the endocytic pathway and for the topologically related processes of viral budding and cytokinesis, but how they accomplish this remains unclear. Using deep-etch electron microscopy, we find that endogenous ESCRT-III filaments stabilized by depleting cells of Vps4 create uniform membrane-deforming conical spirals which are assemblies of specific ESCRT-III heteropolymers. To explore functional roles for ESCRT-III filaments, we examine HIV-1 Gag-mediated budding of virus-like particles and find that depleting Vps4 traps ESCRT-III filaments around nascent Gag assemblies. Interpolating between the observed structures suggests a new role for Vps4 in separating ESCRT-III from Gag or other cargo to allow centripetal growth of a neck constricting ESCRT-III spiral.


Subject(s)
Endosomal Sorting Complexes Required for Transport/chemistry , HIV-1/physiology , Virus Release , Animals , Biological Transport , COS Cells , Cell Membrane/metabolism , Chlorocebus aethiops , Cytokinesis , Cytoplasm/metabolism , Gene Products, gag/chemistry , HEK293 Cells , HIV-1/chemistry , HeLa Cells , Humans , Microscopy, Electron , Microscopy, Fluorescence , Polymers/chemistry , Protein Conformation
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