Hrs and STAM Function Synergistically to Bind Ubiquitin-Modified Cargoes In Vitro

摘要: The turnover of integral membrane proteins requires a specialized transport pathway mediated by components the endosomal sorting complex required for (ESCRT) machinery. In most cases, entry into this that cargoes undergo ubiquitin-modification, thereby facilitating their sequestration on membranes specific, ubiquitin-binding ESCRT subunits. However, requirements underlying initial cargo recognition mono-ubiquitinated cargos remain poorly defined. study, we determine capability each harbors domain to bind reconstituted (VAMP2), which has been covalently linked mono-ubiquitin. We demonstrate ESCRT-0, but not ESCRT-I or ESCRT-II, is able associate stably with within lipid bilayer. Moreover, show domains in both Hrs and STAM must be intact enable binding. These results indicate two subunits ESCRT-0 function together sequester downstream intralumenal vesicles.