SNX3-retromer requires an evolutionary conserved MON2:DOPEY2:ATP9A complex to mediate Wntless sorting and Wnt secretion

TY - JOUR

T1 - SNX3-retromer requires an evolutionary conserved MON2:DOPEY2:ATP9A complex to mediate Wntless sorting and Wnt secretion

AU - McGough, Ian J

AU - de Groot, Reinoud E A

AU - Jellett, Adam P

AU - Betist, Marco C

AU - Varandas, Katherine C

AU - Danson, Chris M

AU - Heesom, Kate J

AU - Korswagen, Hendrik C

AU - Cullen, Peter J

PY - 2018/9/13

Y1 - 2018/9/13

N2 - Wntless transports Wnt morphogens to the cell surface and is required for Wnt secretion and morphogenic gradients formation. Recycling of endocytosed Wntless requires the sorting nexin-3 (SNX3)-retromer-dependent endosome-to-Golgi transport pathway. Here we demonstrate the essential role of SNX3-retromer assembly for Wntless transport and report that SNX3 associates with an evolutionary conserved endosome-associated membrane re-modelling complex composed of MON2, DOPEY2 and the putative aminophospholipid translocase, ATP9A. In vivo suppression of Ce-mon-2, Ce-pad-1 or Ce-tat-5 (respective MON2, DOPEY2 and ATP9A orthologues) phenocopy a loss of SNX3-retromer function, leading to enhanced lysosomal degradation of Wntless and a Wnt phenotype. Perturbed Wnt signalling is also observed upon overexpression of an ATPase-inhibited TAT-5(E246Q) mutant, suggesting a role for phospholipid flippase activity during SNX3-retromer-mediated Wntless sorting. Together, these findings provide in vitro and in vivo mechanistic details to describe SNX3-retromer-mediated transport during Wnt secretion and the formation of Wnt-morphogenic gradients.

AB - Wntless transports Wnt morphogens to the cell surface and is required for Wnt secretion and morphogenic gradients formation. Recycling of endocytosed Wntless requires the sorting nexin-3 (SNX3)-retromer-dependent endosome-to-Golgi transport pathway. Here we demonstrate the essential role of SNX3-retromer assembly for Wntless transport and report that SNX3 associates with an evolutionary conserved endosome-associated membrane re-modelling complex composed of MON2, DOPEY2 and the putative aminophospholipid translocase, ATP9A. In vivo suppression of Ce-mon-2, Ce-pad-1 or Ce-tat-5 (respective MON2, DOPEY2 and ATP9A orthologues) phenocopy a loss of SNX3-retromer function, leading to enhanced lysosomal degradation of Wntless and a Wnt phenotype. Perturbed Wnt signalling is also observed upon overexpression of an ATPase-inhibited TAT-5(E246Q) mutant, suggesting a role for phospholipid flippase activity during SNX3-retromer-mediated Wntless sorting. Together, these findings provide in vitro and in vivo mechanistic details to describe SNX3-retromer-mediated transport during Wnt secretion and the formation of Wnt-morphogenic gradients.

KW - Animals

KW - Biological Transport

KW - Caenorhabditis elegans

KW - Endosomes/metabolism

KW - Golgi Apparatus/metabolism

KW - Green Fluorescent Proteins/metabolism

KW - HEK293 Cells

KW - HeLa Cells

KW - Humans

KW - Mutation

KW - Phenotype

KW - Phospholipid Transfer Proteins/metabolism

KW - Protein Binding

KW - Protein Domains

KW - Proteomics

KW - Proton-Translocating ATPases/metabolism

KW - RNA Interference

KW - Sorting Nexins/metabolism

KW - Transgenes

KW - Vesicular Transport Proteins/metabolism

KW - Wnt Proteins/metabolism

U2 - 10.1038/s41467-018-06114-3

DO - 10.1038/s41467-018-06114-3

M3 - Article

C2 - 30213940

SN - 2041-1723

VL - 9

SP - 3737

JO - Nature Communications

JF - Nature Communications

IS - 1

ER -