title: The Sla2p talin domain plays a role in endocytosis in Saccharomyces cerevisiae.
abstract: Clathrin-binding adaptors play critical roles for endocytosis in multicellular organisms, but their roles in budding yeast have remained unclear. To address this question, we created a quadruple mutant yeast strain lacking the genes encoding the candidate clathrin adaptors Yap1801p, Yap1802p, and Ent2p and containing a truncated version of Ent1p, Ent1DeltaCBMp, missing its clathrin-binding motif. This strain was viable and competent for endocytosis, suggesting the existence of other redundant adaptor-like factors. To identify these factors, we mutagenized the quadruple clathrin adaptor mutant strain and selected cells that were viable in the presence of full-length Ent1p, but inviable with only Ent1DeltaCBMp; these strains were named Rcb (requires clathrin binding). One mutant strain, rcb432, contained a mutation in SLA2 that resulted in lower levels of a truncated protein lacking the F-actin binding talin homology domain. Analyses of this sla2 mutant showed that the talin homology domain is required for endocytosis at elevated temperature, that SLA2 exhibits genetic interactions with both ENT1 and ENT2, and that the clathrin adaptors and Sla2p together regulate the actin cytoskeleton and revealed conditions under which Yap1801p and Yap1802p contribute to viability. Together, our data support the view that Sla2p is an adaptor that links actin to clathrin and endocytosis.
authors.affiliation: Department of Biology, The Johns Hopkins University, Baltimore, Maryland 21218, USA.
authors.forename: Jennifer J
authors.initials: JJ
authors.lastname: Baggett
authors.forename: Katharine E
authors.initials: KE
authors.lastname: D'Aquino
authors.forename: Beverly
authors.initials: B
authors.lastname: Wendland
mesh_terms: D000199:Actins / Q000378:metabolism; D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002966:Clathrin / Q000378:metabolism; D003034:Coated Pits, Cell-Membrane; D003598:Cytoskeletal Proteins; D003599:Cytoskeleton / Q000378:metabolism; D004705:Endocytosis; D009154:Mutation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D016608:Talin / Q000378:metabolism*; D033921:Vesicular Transport Proteins
chemical_list: D000199:Actins; D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport; D002352:Carrier Proteins; D002966:Clathrin; D003598:Cytoskeletal Proteins; C121319:ENT1 protein, S cerevisiae; C121321:ENT2 protein, S cerevisiae; C082266:SLA2 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D016608:Talin; D033921:Vesicular Transport Proteins