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PMID: 10512884 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The Saccharomyces cerevisiae homologue of human Wiskott-Aldrich syndrome protein Las17p interacts with the Arp2/3 complex.

Molecular biology of the cell ·Vol. 10 ·No. 10 ·1999-10-00 ·Pages 3521-38

Madania A, Dumoulin P, Grava S, Kitamoto H, Schärer-Brodbeck C, Soulard A, Moreau V, Winsor B

Abstract

Yeast Las17 protein is homologous to the Wiskott-Aldrich Syndrome protein, which is implicated in severe immunodeficiency. Las17p/Bee1p has been shown to be important for actin patch assembly and actin polymerization. Here we show that Las17p interacts with the Arp2/3 complex. LAS17 is an allele-specific multicopy suppressor of ARP2 and ARP3 mutations; overexpression restores both actin patch organization and endocytosis defects in ARP2 temperature-sensitive (ts) cells. Six of seven ARP2 ts mutants and at least one ARP3 ts mutant are synthetically lethal with las17Delta ts confirming functional interaction with the Arp2/3 complex. Further characterization of las17Delta cells showed that receptor-mediated internalization of alpha factor by the Ste2 receptor is severely defective. The polarity of normal bipolar bud site selection is lost. Las17-gfp remains localized in cortical patches in vivo independently of polymerized actin and is required for the polarized localization of Arp2/3 as well as actin. Coimmunoprecipitation of Arp2p with Las17p indicates that Las17p interacts directly with the complex. Two hybrid results also suggest that Las17p interacts with actin, verprolin, Rvs167p and several other proteins including Src homology 3 (SH3) domain proteins, suggesting that Las17p may integrate signals from different regulatory cascades destined for the Arp2/3p complex and the actin cytoskeleton.

MeSH Terms
Actin-Related Protein 2 Actin-Related Protein 3 Actins/genetics,metabolism Cytoskeletal Proteins Endocytosis/genetics Fungal Proteins/genetics,metabolism Gene Expression Regulation, Fungal Green Fluorescent Proteins Humans Luminescent Proteins Mating Factor Microscopy, Fluorescence Mutation Peptides/metabolism Phenotype Precipitin Tests Receptors, Mating Factor Receptors, Peptide/metabolism Recombinant Fusion Proteins Saccharomyces cerevisiae/genetics,metabolism Saccharomyces cerevisiae Proteins Signal Transduction Suppression, Genetic Transcription Factors Wiskott-Aldrich Syndrome/genetics Wiskott-Aldrich Syndrome Protein
Chemicals
ACTR2 protein, human ACTR3 protein, human Actin-Related Protein 2 Actin-Related Protein 3 Actins Cytoskeletal Proteins Fungal Proteins LAS17 protein, S cerevisiae Luminescent Proteins Peptides Receptors, Mating Factor Receptors, Peptide Recombinant Fusion Proteins Saccharomyces cerevisiae Proteins Transcription Factors Wiskott-Aldrich Syndrome Protein Green Fluorescent Proteins Mating Factor
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Madania A
Mécanismes Moléculaires de la Division Cellulaire et du Développement, Unité Propre de Recherche 9005 du Centre National de la Recherche Scientifique, Institut de Biologie Moléculaire et Cellulaire, F-67084 Strasbourg, France.
Dumoulin P
Grava S
Kitamoto H
Schärer-Brodbeck C
Soulard A
Moreau V
Winsor B
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.10.10.3521
Published
1999-10-00
Pages
3521-38
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC25621
Subset
IM
External Links
PubMed source DOI: 10.1091/mbc.10.10.3521 PMC Full Text View journal details
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