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Nakagawa Lab

Mammalian Retromer Is an Adaptable Scaffold for Cargo Sorting from Endosomes

AUTHORS

Kendall AKAmy K , Xie BBoyang , Xu PPeng , Wang JJue , Burcham RRodger , Frazier MNMeredith N , Binshtein EElad , Wei HHui , Graham TRTodd R , Nakagawa TTerunaga , Jackson LPLauren P . Structure (London, England : 1993). 2020 02 04; ().

ABSTRACT

Metazoan retromer (VPS26/VPS35/VPS29) associates with sorting nexins on endosomal tubules to sort proteins to the trans-Golgi network or plasma membrane. Mechanisms of metazoan retromer assembly remain undefined. We combine single-particle cryoelectron microscopy with biophysical methods to uncover multiple oligomer structures. 2D class averages reveal mammalian heterotrimers; dimers of trimers; tetramers of trimers; and flat chains. These species are further supported by biophysical solution studies. We provide reconstructions of all species, including key sub-structures (∼5 Å resolution). Local resolution variation suggests that heterotrimers and dimers adopt multiple conformations. Our structures identify a flexible, highly conserved electrostatic dimeric interface formed by VPS35 subunits. We generate structure-based mutants to disrupt this interface in vitro. Equivalent mutations in yeast demonstrate a mild cargo-sorting defect. Our data suggest the metazoan retromer is an adaptable and plastic scaffold that accommodates interactions with different sorting nexins to sort multiple cargoes from endosomes their final destinations.