title: The yeast Epsin Ent1 is recruited to membranes through multiple independent interactions.
abstract: In addition to its well known role in targeting proteins for proteasomal degradation, ubiquitin (Ub) is also involved in promoting internalization of cell surface proteins into the endocytic pathway. Moreover, putative Ub interaction motifs (UIMs) as well as Ub-associated (UBA) domains have been identified in key yeast endocytic proteins (the epsins Ent1 and Ent2, and the Eps15 homolog Ede1). In this study, we characterized the interaction of Ub with the Ede1 UBA domain and with the UIMs of Ent1. Our data suggest that the UIMs and the UBA are involved in binding these proteins to biological membranes. We also show that the Ent1 ENTH domain binds to phosphoinositides in vitro and that Ent1 NPF motifs interact with the EH domain-containing proteins Ede1 and Pan1. Our findings indicate that the ENTH domain interaction with membrane lipids cooperates with the binding of membrane-associated Ub moieties. These events may in turn favor the occurrence of other interactions, for instance EH-NPF recognition, thus stabilizing networks of low affinity binding partners at endocytic sites.
authors.affiliation: Department of Biology, The Johns Hopkins University, Baltimore, Maryland 21218, USA.
authors.forename: Rubén Claudio
authors.initials: RC
authors.lastname: Aguilar
authors.forename: Hadiya A
authors.initials: HA
authors.lastname: Watson
authors.forename: Beverly
authors.initials: B
authors.lastname: Wendland
mesh_terms: D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D004705:Endocytosis; D005656:Fungal Proteins / Q000378:metabolism*; D008958:Models, Molecular; D010716:Phosphatidylinositols / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D025801:Ubiquitin / Q000378:metabolism; D033921:Vesicular Transport Proteins
chemical_list: D002352:Carrier Proteins; C121319:ENT1 protein, S cerevisiae; D005656:Fungal Proteins; D010716:Phosphatidylinositols; D029701:Saccharomyces cerevisiae Proteins; D025801:Ubiquitin; D033921:Vesicular Transport Proteins