Higher‐order assemblies of oligomeric cargo receptor complexes form the membrane scaffold of the Cvt vesicle

Article Article scientifique Article Higher‐order assemblies of oligomeric cargo receptor complexes form the membrane scaffold of the Cvt vesicle eng 2016 author Bertipaglia Chiara author Schneider Sarah 856190 author Jakobi Arjen J author Tarafder Abul K author Bykov Yury S author Picco Andrea 842776 author Kukulski Wanda author Kosinski Jan author Hagen Wim JH author Ravichandran Arvind C author Wilmanns Matthias author Kaksonen Marko 0000-0003-3645-7689 830048 author Briggs John AG author Sachse Carsten 1044-1060 10.15252/embr.201541960 1469-221X https://onlinelibrary.wiley.com/doi/abs/10.15252/embr.201541960 Selective autophagy is the mechanism by which large cargos are specifically sequestered for degradation. The structural details of cargo and receptor assembly giving rise to autophagic vesicles remain to be elucidated. We utilize the yeast cytoplasm‐to‐vacuole targeting (Cvt) pathway, a prototype of selective autophagy, together with a multi‐scale analysis approach to study the molecular structure of Cvt vesicles. We report the oligomeric nature of the major Cvt cargo Ape1 with a combined 2.8 Å X‐ray and negative stain EM structure, as well as the secondary cargo Ams1 with a 6.3 Å cryo‐EM structure. We show that the major dodecameric cargo prApe1 exhibits a tendency to form higher‐order chain structures that are broken upon interaction with the receptor Atg19 in vitro. The stoichiometry of these cargo–receptor complexes is key to maintaining the size of the Cvt aggregate in vivo. Using correlative light and electron microscopy, we further visualize key stages of Cvt vesicle biogenesis. Our findings suggest that Atg19 interaction limits Ape1 aggregate size while serving as a vehicle for vacuolar delivery of tetrameric Ams1. Autophagy cargo Autophagy receptor Correlative light and electron microscopy Electron cryomicroscopy Fluorescence light microscopy Selective autophagy X‐ray crystallography Dewey 540 39 Département de biochimie 144 false EMBO Reports 17 7 fe018e57-2243-46bd-a17a-a67ebc1bd243 Article (Published version) Public Higher_MK_16.pdf 3079895 application/pdf openAccess Faculté des sciences / Section de chimie et biochimie / Département de biochimie

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Higher‐order assemblies of oligomeric cargo receptor complexes form the membrane scaffold of the Cvt vesicle	EMBO Reports			2016	257	116

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Schneider, Sarah