Concerted ESCRT and clathrin recruitment waves define the timing and morphology of intraluminal vesicle formation

Eva Maria Wenzel (), Sebastian Wolfgang Schultz, Kay Oliver Schink, Nina Marie Pedersen, Viola Nähse, Andreas Carlson, Andreas Brech, Harald Stenmark and Camilla Raiborg ()
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Eva Maria Wenzel: Oslo University Hospital, Montebello
Sebastian Wolfgang Schultz: Oslo University Hospital, Montebello
Kay Oliver Schink: Oslo University Hospital, Montebello
Nina Marie Pedersen: Oslo University Hospital, Montebello
Viola Nähse: Oslo University Hospital, Montebello
Andreas Carlson: University of Oslo
Andreas Brech: Oslo University Hospital, Montebello
Harald Stenmark: Oslo University Hospital, Montebello
Camilla Raiborg: Oslo University Hospital, Montebello

Nature Communications, 2018, vol. 9, issue 1, 1-18

Abstract: Abstract The endosomal sorting complex required for transport (ESCRT) machinery mediates cargo sorting, membrane deformation and membrane scission on the surface of endosomes, generating intraluminal vesicles (ILVs) to degrade signaling receptors. By live-cell imaging of individual endosomes in human cells, we find that ESCRT proteins are recruited in a repetitive pattern: ESCRT-0 and -I show a gradual and linear recruitment and dissociation, whereas ESCRT-III and its regulatory ATPase VPS4 display fast and transient dynamics. Electron microscopy shows that ILVs are formed consecutively, starting immediately after endocytic uptake of cargo proteins and correlating with the repeated ESCRT recruitment waves, unraveling the timing of ILV formation. Clathrin, recruited by ESCRT-0, is required for timely ESCRT-0 dissociation, efficient ILV formation, correct ILV size and cargo degradation. Thus, cargo sorting and ILV formation occur by concerted, coordinated and repetitive recruitment waves of individual ESCRT subcomplexes and are controlled by clathrin.

Date: 2018
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DOI: 10.1038/s41467-018-05345-8

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