HRS-WASH axis governs actin-mediated endosomal recycling and cell invasion.

Ewan Macdonald,Daniel Newman,Michael J Clague,Jessica Bithell,Louise Brown,Douglas Grimes,Tobias Zech,Sylvie Urbé,Han Liu,Arnaud Selvais,Thomas Waring

doi:10.1083/jcb.201710051

Abstract

Transmembrane proteins in the sorting endosome are either recycled to their point of origin or destined for lysosomal degradation. Lysosomal sorting is mediated by interaction of ubiquitylated transmembrane proteins with the endosomal sorting complex required for transport (ESCRT) machinery. In this study, we uncover an alternative role for the ESCRT-0 component hepatocyte growth factor-regulated tyrosine kinase substrate (HRS) in promoting the constitutive recycling of transmembrane proteins. We find that endosomal localization of the actin nucleating factor Wiscott-Aldrich syndrome protein and SCAR homologue (WASH) requires HRS, which occupies adjacent endosomal subdomains. Depletion of HRS results in defective constitutive recycling of epidermal growth factor receptor and the matrix metalloproteinase MT1-MMP, leading to their accumulation in internal compartments. We show that direct interactions with endosomal actin are required for efficient recycling and use a model system of chimeric transferrin receptor trafficking to show that an actin-binding motif can counteract an ubiquitin signal for lysosomal sorting. Directed receptor recycling is used by cancer cells to achieve invasive migration. Accordingly, abrogating HRS- and actin-dependent MT1-MMP recycling results in defective matrix degradation and invasion of triple-negative breast cancer cells.

Highlights

Cell surface proteins that enter endosomes may be recycled to the plasma membrane or otherwise actively sorted toward the lysosomal pathway
We identify the function of the intrinsic actin-binding domains (ABDs) of EGF receptor (EGFR) and membrane type 1–matrix metalloproteinase (MT1-MMP) as directing the default sorting to recycling under steady-state conditions and show that hepatocyte growth factor–regulated tyrosine kinase substrate (HRS)-orchestrated endosomal actin is required for MT1-MMP–dependent invasive cell migration of breast cancer cells
HRS occupies a separate subdomain but is required for the recruitment of Wiscott-Aldrich syndrome protein and SCAR homologue (WASH) to endosomes Costaining of fixed cells with WASH and HRS antibodies revealed a high degree of colocalization in HeLa and MDA–MB-231 cells

Summary

Introduction

Cell surface proteins that enter endosomes may be recycled to the plasma membrane or otherwise actively sorted toward the lysosomal pathway. A more complex feature of the recycling pathway is represented by its ability to distribute to different regions of the cell, for example the leading edge of migrating cells or one or the other membrane of polarized cells (Matter and Mellman, 1994). Such recycling of membrane type 1–matrix metalloproteinase (MT1-MMP) and EGF receptor (EGFR) drives cancer cell invasion (Caswell et al, 2008; Steffen et al, 2008)

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