Pkh1/2-dependent phosphorylation of Vps27 regulates ESCRT-I recruitment to endosomes.

Mol Biol Cell 2012 Oct 23;23(20):4054-64. Epub 2012 Aug 23.

Department of Molecular and Cellular Genetics, Unité Mixte de Recherche 7156, Université de Strasbourg and Centre National de la Recherche Scientifique, 67084 Strasbourg, France.

Multivesicular endosomes (MVBs) are major sorting platforms for membrane proteins and participate in plasma membrane protein turnover, vacuolar/lysosomal hydrolase delivery, and surface receptor signal attenuation. MVBs undergo unconventional inward budding, which results in the formation of intraluminal vesicles (ILVs). MVB cargo sorting and ILV formation are achieved by the concerted function of endosomal sorting complex required for transport (ESCRT)-0 to ESCRT-III. The ESCRT-0 subunit Vps27 is a key player in this pathway since it recruits the other complexes to endosomes. Here we show that the Pkh1/Phk2 kinases, two yeast orthologues of the 3-phosphoinositide-dependent kinase, phosphorylate directly Vps27 in vivo and in vitro. We identify the phosphorylation site as the serine 613 and demonstrate that this phosphorylation is required for proper Vps27 function. Indeed, in pkh-ts temperature-sensitive mutant cells and in cells expressing vps27(S613A), MVB sorting of the carboxypeptidase Cps1 and of the α-factor receptor Ste2 is affected and the Vps28-green fluorescent protein ESCRT-I subunit is mainly cytoplasmic. We propose that Vps27 phosphorylation by Pkh1/2 kinases regulates the coordinated cascade of ESCRT complex recruitment at the endosomal membrane.

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http://dx.doi.org/10.1091/mbc.E12-01-0001DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3469520PMC
October 2012
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