Ent5p is required with Ent3p and Vps27p for ubiquitin-dependent protein sorting into the multivesicular body.
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.
Eugster
Anne
A
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.
Pécheur
Eve-Isabelle
EI
Michel
Fabrice
F
Winsor
Barbara
B
Letourneur
François
F
Friant
Sylvie
S
D033942:Adaptor Proteins, Vesicular Transport / Q000032:analysis / Q000378:metabolism / Q000502:physiology*; D000595:Amino Acid Sequence; D002268:Carboxypeptidases / Q000032:analysis; D004705:Endocytosis; D056827:Endosomal Sorting Complexes Required for Transport; D011992:Endosomes / Q000276:immunology / Q000378:metabolism*; D047468:Immunoprecipitation; D008969:Molecular Sequence Data; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000032:analysis / Q000378:metabolism; D021381:Protein Transport; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis / Q000378:metabolism* / Q000502:physiology*; D016415:Sequence Alignment; D025801:Ubiquitin / Q000378:metabolism; D033921:Vesicular Transport Proteins / Q000032:analysis / Q000378:metabolism*
D033942:Adaptor Proteins, Vesicular Transport; D056827:Endosomal Sorting Complexes Required for Transport; C495587:Ent3 protein, S cerevisiae; C495588:Ent5 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D025801:Ubiquitin; C119627:VPS27 protein, S cerevisiae; D033921:Vesicular Transport Proteins; C095504:FAB1 protein, S cerevisiae; D017853:Phosphotransferases (Alcohol Group Acceptor); D002268:Carboxypeptidases; C019836:CPS1 protein, S cerevisiae