PT - JOURNAL ARTICLE AU - Daniel S. Johnson AU - Marina Bleck AU - Sanford M. Simon TI - Timing of ESCRT-III protein recruitment and membrane scission during HIV-1 assembly AID - 10.1101/281774 DP - 2018 Jan 01 TA - bioRxiv PG - 281774 4099 - http://biorxiv.org/content/early/2018/03/13/281774.short 4100 - http://biorxiv.org/content/early/2018/03/13/281774.full AB - 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 seconds before scission. These observations suggest 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-III 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.