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

被引:19
作者
Bertipaglia, Chiara [1 ]
Schneider, Sarah [1 ]
Jakobi, Arjen J. [1 ,2 ]
Tarafder, Abul K. [1 ]
Bykov, Yury S. [1 ]
Picco, Andrea [3 ,4 ]
Kukulski, Wanda [1 ,3 ,5 ]
Kosinski, Jan [1 ]
Hagen, Wim J. H. [1 ]
Ravichandran, Arvind C. [1 ]
Wilmanns, Matthias [2 ]
Kaksonen, Marko [3 ,4 ]
Briggs, John A. G. [1 ,3 ]
Sachse, Carsten [1 ]
机构
[1] European Mol Biol Lab, Struct & Computat Biol Unit, Heidelberg, Germany
[2] European Mol Biol Lab, Hamburg Unit, Hamburg, Germany
[3] European Mol Biol Lab, Cell Biol & Biophys Unit, Heidelberg, Germany
[4] Univ Geneva, Dept Biochem, Geneva, Switzerland
[5] MRC, Mol Biol Lab, Cell Biol, Cambridge, England
基金
瑞士国家科学基金会;
关键词
autophagy cargo; autophagy receptor; correlative light and electron microscopy; electron cryomicroscopy; fluorescence light microscopy; selective autophagy; X-ray crystallography; YEAST AMINOPEPTIDASE-I; SELECTIVE AUTOPHAGY; SACCHAROMYCES-CEREVISIAE; CORRELATED FLUORESCENCE; STRUCTURE PREDICTION; QUATERNARY STRUCTURE; ALPHA-MANNOSIDASE; ATG PROTEINS; VACUOLE; CYTOPLASM;
D O I
10.15252/embr.201541960
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 angstrom X-ray and negative stain EM structure, as well as the secondary cargo Ams1 with a 6.3 angstrom 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.
引用
收藏
页码:1044 / 1060
页数:17
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