In vitro reconstitution of calcium-dependent recruitment of the human ESCRT machinery in lysosomal membrane repair.

ABSTRACT: The endosomal sorting complex required for transport (ESCRT) machinery is centrally involved in the repair of damage to both the plasma and lysosome membranes. ESCRT recruitment to sites of damage occurs on a fast time scale, and Ca²⁺ has been proposed to play a key signaling role in the process. Here, we show that the Ca²⁺-binding regulatory protein ALG-2 binds directly to negatively charged membranes in a Ca²⁺-dependent manner. Next, by monitoring the colocalization of ALIX with ALG-2 on negatively charged membranes, we show that ALG-2 recruits ALIX to the membrane. Furthermore, we show that ALIX recruitment to the membrane orchestrates the downstream assembly of late-acting CHMP4B, CHMP3, and CHMP2A subunits along with the AAA⁺ ATPase VPS4B. F

SUBMITTER: Shukla S 

PROVIDER: S-EPMC9436306 | biostudies-literature | 2022 Aug

REPOSITORIES: biostudies-literature