Inhibition of ESCRT-II-CHMP6 interactions impedes cytokinetic abscission and leads to cell death.
Bottom Line: This phenotype is abolished in a mutated version of CHMP6-N designed to prevent CHMP6-N binding to its ESCRT-II partner.Of interest, deleting the first 10 amino acids from CHMP6-N does not interfere with its arrival at the intercellular bridge but almost completely abolishes the abscission failure phenotype.Our work advances the mechanistic understanding of ESCRT-mediated membrane fission in cells and introduces an easily applicable tool for upstream inhibition of the ESCRT pathway in live mammalian cells.
Affiliation: Department of Life Sciences and the National Institute for Biotechnology in the Negev (NIBN), Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel.Show MeSH
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Mentions: We next examined the spatiotemporal organization of the ESCRT-II complex during cytokinesis. Live-cell imaging revealed that the ESCRT-II components VPS36 and VPS22 localize to the intercellular bridge during late cytokinesis (Figure 2, A and B, Supplemental Figure S1A, and Supplemental Videos S2–S4, respectively). Quantitative analysis of the recruitment pattern of VPS36 to the intercellular bridge revealed that VPS36 begins accumulating at the bridge only ∼20 min before final scission and reaches its peak levels at the time of scission (Figure 2A and Supplemental Video S2). The increase in VPS36 levels is temporally correlated with acute constriction of the microtubules on either side of the intercellular bridge (Figure 2B and Supplemental Video S3). VPS36 was also clearly observed in postabscission remnants (Figure 2B and Supplemental Video S3).
Affiliation: Department of Life Sciences and the National Institute for Biotechnology in the Negev (NIBN), Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel.