Oral Presentation The 35th Biennial Conference of the Society of Crystallographers in Australia and New Zealand 2024 (Crystal 35)

Structural basis for coupling of the WASH subunit FAM21 with the endosomal SNX27-Retromer complex (108954)

Qian Guo 1 , Kai-En Chen 1 , Manuel Gimenez-Andres 2 , Peter Cullen 2 , Brett Collins 1
  1. The University of Queensland, St Lucia, QLD, Australia
  2. School of Biochemistry, University of Bristol, Bristol, UK

Endosomal membrane trafficking is mediated by specific protein coats and formation of actin-rich membrane domains. The Retromer complex coordinates with sorting nexin (SNX) cargo adaptors including SNX27, and the SNX27–Retromer assembly interacts with the WASH complex which nucleates actin filaments establishing the endosomal reycling domain. Crystal structures, modelling, biochemical and cellular validation reveal how the FAM21 subunit of WASH interacts with both Retromer and SNX27. FAM21 binds the FERM domain of SNX27 using acidic-Asp-Leu-Phe (aDLF) motifs similar to those found in the SNX1 and SNX2 subunits of the ESCPE-1 complex. Overlapping FAM21 repeats and a specific Pro-Leu containing motif bind three distinct sites on Retromer involving both the VPS35 and VPS29 subunits. Mutation of the major VPS35-binding site does not prevent cargo recycling, however it partially reduces endosomal WASH association indicating that a network of redundant interactions promote endosomal activity of the WASH complex. These studies establish the molecular basis for how SNX27–Retromer is coupled to the WASH complex via overlapping and multiplexed motif-based interactions required for the dynamic assembly of endosomal membrane recycling domains.

 

[1] Structural basis for coupling of the WASH subunit FAM21 with the endosomal SNX27-Retromer complex. Qian Guo, Kai-en Chen, Manuel Gimenez-Andres, Adam P. Jellett, Ya Gao, Boris Simonetti, Meihan Liu, Chris M. Danson, Kate J. Heesom, Peter J. Cullen, Brett M. Collins

bioRxiv 2023.08.15.553351; doi: https://doi.org/10.1101/2023.08.15.553351