Ent5p is required with Ent3p and Vps27p for ubiquitin-dependent protein sorting into the multivesicular body

Anne Eugster; Eve-Isabelle Pécheur; Fabrice Michel; Barbara Winsor; François Letourneur; Sylvie Friant

doi:10.1091/mbc.e03-11-0793

Ent5p is required with Ent3p and Vps27p for ubiquitin-dependent protein sorting into the multivesicular body

Mol Biol Cell. 2004 Jul;15(7):3031-41. doi: 10.1091/mbc.e03-11-0793. Epub 2004 Apr 23.

Authors

Anne Eugster¹, Eve-Isabelle Pécheur, Fabrice Michel, Barbara Winsor, François Letourneur, Sylvie Friant

Affiliation

¹ Laboratoire de Transport et Compartimentation Intracellulaire, Institut de Biologie et Chimie des Protéines, UMR5086 Centre National de la Recherche Scientifique, Institut Fédératif de Recherche 128 BioSciences Lyon-Gerland, 69367 Lyon, France.

Abstract

At the late endosomes, cargoes destined for the interior of the vacuole are sorted into invaginating vesicles of the multivesicular body. Both PtdIns(3,5)P(2) and ubiquitin are necessary for proper sorting of some of these cargoes. We show that Ent5p, a yeast protein of the epsin family homologous to Ent3p, localizes to endosomes and specifically binds to PtdIns(3,5)P(2) via its ENTH domain. In cells lacking Ent3p and Ent5p, ubiquitin-dependent sorting of biosynthetic and endocytic cargo into the multivesicular body is disrupted, whereas other trafficking routes to the vacuole are not affected. Ent3p and Ent5p are associated with Vps27p, a FYVE domain containing protein that interacts with ubiquitinated cargoes and is required for protein sorting into the multivesicular body. Therefore, Ent3p and Ent5p are the first proteins shown to be connectors between PtdIns(3,5)P(2)- and the Vps27p-ubiquitin-driven sorting machinery at the multivesicular body.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adaptor Proteins, Vesicular Transport / analysis
Adaptor Proteins, Vesicular Transport / metabolism
Adaptor Proteins, Vesicular Transport / physiology*
Amino Acid Sequence
Carboxypeptidases / analysis
Endocytosis
Endosomal Sorting Complexes Required for Transport
Endosomes / immunology
Endosomes / metabolism*
Immunoprecipitation
Molecular Sequence Data
Phosphotransferases (Alcohol Group Acceptor) / analysis
Phosphotransferases (Alcohol Group Acceptor) / metabolism
Protein Transport
Saccharomyces cerevisiae / metabolism*
Saccharomyces cerevisiae Proteins / analysis
Saccharomyces cerevisiae Proteins / metabolism*
Saccharomyces cerevisiae Proteins / physiology*
Sequence Alignment
Ubiquitin / metabolism
Vesicular Transport Proteins / analysis
Vesicular Transport Proteins / metabolism*

Substances

Adaptor Proteins, Vesicular Transport
Endosomal Sorting Complexes Required for Transport
Ent3 protein, S cerevisiae
Ent5 protein, S cerevisiae
Saccharomyces cerevisiae Proteins
Ubiquitin
VPS27 protein, S cerevisiae
Vesicular Transport Proteins
FAB1 protein, S cerevisiae
Phosphotransferases (Alcohol Group Acceptor)
Carboxypeptidases
CPS1 protein, S cerevisiae