Ent3p Is a PtdIns(3,5)P2 effector required for protein sorting to the multivesicular body.
PtdIns(3,5)P(2) is required for cargo-selective sorting to the vacuolar lumen via the multivesicular body (MVB). Here we show that Ent3p, a yeast epsin N-terminal homology (ENTH) domain-containing protein, is a specific PtdIns(3,5)P(2) effector localized to endosomes. The ENTH domain of Ent3p is essential for its PtdIns(3,5)P(2) binding activity and for its membrane interaction in vitro and in vivo. Ent3p is required for protein sorting into the MVB but not for the internalization step of endocytosis. Ent3p is associated with clathrin and is necessary for normal actin cytoskeleton organization. Our results show that Ent3p is required for protein sorting into intralumenal vesicles of the MVB through PtdIns(3,5)P(2) binding via its ENTH domain.
Friant
Sylvie
S
Laboratoire de Transport et Compartimentation Intracellulaire, Institut de Biologie et Chimie des Protéines, UMR 5086 CNRS, IFR 128 BioSciences Lyon-Gerland 7, passage du Vercors, 69367, Lyon, France. s.friant@ibcp.fr
Pécheur
Eve Isabelle
EI
Eugster
Anne
A
Michel
Fabrice
F
Lefkir
Yaya
Y
Nourrisson
Delphine
D
Letourneur
François
F
D000199:Actins / Q000378:metabolism; D033942:Adaptor Proteins, Vesicular Transport; D019883:Alkyl and Aryl Transferases / Q000378:metabolism; D015153:Blotting, Western; D002352:Carrier Proteins / Q000378:metabolism*; D002966:Clathrin / Q000378:metabolism; D004705:Endocytosis / Q000502:physiology; D011992:Endosomes / Q000378:metabolism; D005656:Fungal Proteins; D049452:Green Fluorescent Proteins; D066298:In Vitro Techniques; D007700:Kinetics; D008164:Luminescent Proteins / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000378:metabolism; D009097:Multienzyme Complexes / Q000378:metabolism; D009154:Mutation; D009479:Neuropeptides / Q000378:metabolism*; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism*; D011233:Precipitin Tests; D021382:Protein Sorting Signals / Q000502:physiology*; D017434:Protein Structure, Tertiary / Q000502:physiology*; D021381:Protein Transport; D011726:Pyridinium Compounds / Q000378:metabolism; D000644:Quaternary Ammonium Compounds / Q000378:metabolism; D016415:Sequence Alignment / Q000379:methods; D013696:Temperature; D013997:Time Factors; D014617:Vacuoles / Q000378:metabolism*; D033921:Vesicular Transport Proteins; D015003:Yeasts
D000199:Actins; D033942:Adaptor Proteins, Vesicular Transport; D002352:Carrier Proteins; D002966:Clathrin; C092350:FM 4-64; D005656:Fungal Proteins; D008164:Luminescent Proteins; D009097:Multienzyme Complexes; D009479:Neuropeptides; D018129:Phosphatidylinositol Phosphates; D021382:Protein Sorting Signals; D011726:Pyridinium Compounds; D000644:Quaternary Ammonium Compounds; D033921:Vesicular Transport Proteins; C114154:epsin; C106336:phosphatidylinositol 3,5-diphosphate; D049452:Green Fluorescent Proteins; D006899:Mixed Function Oxygenases; C035189:peptidylglycine monooxygenase; D019883:Alkyl and Aryl Transferases; C115307:CPS protein, Gibberella fujikuroi