The Saccharomyces cerevisiae homologue of human Wiskott-Aldrich syndrome protein Las17p interacts with the Arp2/3 complex.
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.
Madania
A
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
P
Grava
S
S
Kitamoto
H
H
Schärer-Brodbeck
C
C
Soulard
A
A
Moreau
V
V
Winsor
B
B
D051377:Actin-Related Protein 2; D051378:Actin-Related Protein 3; D000199:Actins / Q000235:genetics / Q000378:metabolism*; D003598:Cytoskeletal Proteins; D004705:Endocytosis / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins; D000072235:Mating Factor; D008856:Microscopy, Fluorescence; D009154:Mutation; D010455:Peptides / Q000378:metabolism; D010641:Phenotype; D011233:Precipitin Tests; D051964:Receptors, Mating Factor; D018000:Receptors, Peptide / Q000378:metabolism; D011993:Recombinant Fusion Proteins; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D015398:Signal Transduction; D013489:Suppression, Genetic; D014157:Transcription Factors; D014923:Wiskott-Aldrich Syndrome / Q000235:genetics; D051316:Wiskott-Aldrich Syndrome Protein
C495926:ACTR2 protein, human; C495928:ACTR3 protein, human; D051377:Actin-Related Protein 2; D051378:Actin-Related Protein 3; D000199:Actins; D003598:Cytoskeletal Proteins; D005656:Fungal Proteins; C480361:LAS17 protein, S cerevisiae; D008164:Luminescent Proteins; D010455:Peptides; D051964:Receptors, Mating Factor; D018000:Receptors, Peptide; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D051316:Wiskott-Aldrich Syndrome Protein; D049452:Green Fluorescent Proteins; D000072235:Mating Factor