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Published 12 April 2004. doi:10.1083/jcb.200312034
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 165, Number 1, 111-122
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Article

 Cargo-selective endosomal sorting for retrieval to the Golgi requires retromer

Matthew N.J. Seaman

Cambridge Institute for Medical Research and Department of Clinical Biochemistry, University of Cambridge, Cambridge CB2 2XY, England, UK

Address correspondence to M.N.J. Seaman, Cambridge Institute for Medical Research, University of Cambridge, Wellcome Trust/MRC Building, Addenbrookes Hospital, Hills Road, Cambridge CB2 2XY, England, UK. Tel.: (44) 1223-762627. Fax: (44) 1223-762640. email: mnjs100{at}cam.ac.uk

fEndosome-to-Golgi retrieval of the mannose 6-phosphate receptor (MPR) is required for lysosome biogenesis. Currently, this pathway is poorly understood. Analyses in yeast identified a complex of proteins called "retromer" that is essential for endosome-to-Golgi retrieval of the carboxypeptidase Y receptor Vps10p. Retromer comprises five distinct proteins: Vps35p, 29p, 26p, 17p, and 5p, which are conserved in mammals. Here, we show that retromer is required for the efficient retrieval of the cation-independent MPR (CI-MPR). Cells lacking mammalian VPS26 fail to retrieve the CI-MPR, resulting in either rapid degradation of or mislocalization to the plasma membrane. We have localized mVPS26 to multivesicular body endosomes by electron microscopy, and through the use of CD8 reporter protein constructs have examined the effect of loss of mVPS26 upon the trafficking of membrane proteins that cycle between the endosome and the Golgi. The data presented here support the hypothesis that retromer performs a selective function in endosome-to-Golgi transport, mediating retrieval of the CI-MPR, but not furin.

Key Words: retromer; vesicle; retrieval; endosome; Golgi

The online version of this article includes supplemental material.

Abbreviations used in this paper: CD-MPR, cation-dependent mannose 6-phosphate receptor; CI-MPR, cation-independent mannose 6-phosphate receptor; CPY, carboxypeptidase Y; MPR, mannose 6-phosphate receptor; siRNA, small interfering RNA; Snx, sorting nexin; TfnR, transferrin receptor.


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