Timing of ESCRT-III protein recruitment and membrane scission during HIV-1 assembly

Elife. 2018 Jul 4:7:e36221. doi: 10.7554/eLife.36221.

Abstract

The Endosomal Sorting Complexes Required for Transport III (ESCRT-III) proteins are critical for cellular membrane scission processes with topologies inverted relative to clathrin-mediated endocytosis. Some viruses appropriate ESCRT-IIIs for their release. By imaging single assembling viral-like particles of HIV-1, we observed that ESCRT-IIIs and the ATPase VPS4 arrive after most of the virion membrane is bent, linger for tens of seconds, and depart ~20 s before scission. These observations suggest that ESCRT-IIIs are recruited by a combination of membrane curvature and the late domains of the HIV-1 Gag protein. ESCRT-IIIs may pull the neck into a narrower form but must leave to allow scission. If scission does not occur within minutes of ESCRT departure, ESCRT-IIIs and VPS4 are recruited again. This mechanistic insight is likely relevant for other ESCRT-dependent scission processes including cell division, endosome tubulation, multivesicular body and nuclear envelope formation, and secretion of exosomes and ectosomes.

Keywords: ESCRT; HIV-1; cell biology; human; infectious disease; membrane scission; microbiology; viral assembly.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cell Membrane / metabolism*
  • Computer Simulation
  • Cytosol / metabolism
  • Endosomal Sorting Complexes Required for Transport / metabolism*
  • Gene Knockdown Techniques
  • Green Fluorescent Proteins / metabolism
  • HEK293 Cells
  • HIV-1 / physiology*
  • HeLa Cells
  • Humans
  • Hydrogen-Ion Concentration
  • Models, Biological
  • RNA, Small Interfering / metabolism
  • Time Factors
  • Vacuolar Proton-Translocating ATPases / metabolism
  • Virion / metabolism
  • Virus Assembly / physiology*
  • gag Gene Products, Human Immunodeficiency Virus / chemistry
  • gag Gene Products, Human Immunodeficiency Virus / metabolism

Substances

  • Endosomal Sorting Complexes Required for Transport
  • PHluorin
  • RNA, Small Interfering
  • gag Gene Products, Human Immunodeficiency Virus
  • Green Fluorescent Proteins
  • Vacuolar Proton-Translocating ATPases