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Effects of Arp2 and Arp3 nucleotide-binding pocket mutations on Arp2/3 complex function

Contributions of actin-related proteins (Arp) 2 and 3 nucleotide state to Arp2/3 complex function were tested using nucleotide-binding pocket (NBP) mutants in Saccharomyces cerevisiae. ATP binding by Arp2 and Arp3 was required for full Arp2/3 complex nucleation activity in vitro. Analysis of actin d...

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Autores principales: Martin, Adam C., Xu, Xiao-Ping, Rouiller, Isabelle, Kaksonen, Marko, Sun, Yidi, Belmont, Lisa, Volkmann, Niels, Hanein, Dorit, Welch, Matthew, Drubin, David G.
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2005
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2171590/
https://www.ncbi.nlm.nih.gov/pubmed/15657399
http://dx.doi.org/10.1083/jcb.200408177
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author Martin, Adam C.
Xu, Xiao-Ping
Rouiller, Isabelle
Kaksonen, Marko
Sun, Yidi
Belmont, Lisa
Volkmann, Niels
Hanein, Dorit
Welch, Matthew
Drubin, David G.
author_facet Martin, Adam C.
Xu, Xiao-Ping
Rouiller, Isabelle
Kaksonen, Marko
Sun, Yidi
Belmont, Lisa
Volkmann, Niels
Hanein, Dorit
Welch, Matthew
Drubin, David G.
author_sort Martin, Adam C.
collection PubMed
description Contributions of actin-related proteins (Arp) 2 and 3 nucleotide state to Arp2/3 complex function were tested using nucleotide-binding pocket (NBP) mutants in Saccharomyces cerevisiae. ATP binding by Arp2 and Arp3 was required for full Arp2/3 complex nucleation activity in vitro. Analysis of actin dynamics and endocytosis in mutants demonstrated that nucleotide-bound Arp3 is particularly important for Arp2/3 complex function in vivo. Severity of endocytic defects did not correlate with effects on in vitro nucleation activity, suggesting that a critical Arp2/3 complex function during endocytosis may be structural rather than catalytic. A separate class of Arp2 and Arp3 NBP mutants suppressed phenotypes of mutants defective for actin nucleation. An Arp2 suppressor mutant increased Arp2/3 nucleation activity. Electron microscopy of Arp2/3 complex containing this Arp2 suppressor identified a structural change that also occurs upon Arp2/3 activation by nucleation promoting factors. These data demonstrate the importance of Arp2 and Arp3 nucleotide binding for nucleating activity, and Arp3 nucleotide binding for maintenance of cortical actin cytoskeleton cytoarchitecture.
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spelling pubmed-21715902008-03-05 Effects of Arp2 and Arp3 nucleotide-binding pocket mutations on Arp2/3 complex function Martin, Adam C. Xu, Xiao-Ping Rouiller, Isabelle Kaksonen, Marko Sun, Yidi Belmont, Lisa Volkmann, Niels Hanein, Dorit Welch, Matthew Drubin, David G. J Cell Biol Research Articles Contributions of actin-related proteins (Arp) 2 and 3 nucleotide state to Arp2/3 complex function were tested using nucleotide-binding pocket (NBP) mutants in Saccharomyces cerevisiae. ATP binding by Arp2 and Arp3 was required for full Arp2/3 complex nucleation activity in vitro. Analysis of actin dynamics and endocytosis in mutants demonstrated that nucleotide-bound Arp3 is particularly important for Arp2/3 complex function in vivo. Severity of endocytic defects did not correlate with effects on in vitro nucleation activity, suggesting that a critical Arp2/3 complex function during endocytosis may be structural rather than catalytic. A separate class of Arp2 and Arp3 NBP mutants suppressed phenotypes of mutants defective for actin nucleation. An Arp2 suppressor mutant increased Arp2/3 nucleation activity. Electron microscopy of Arp2/3 complex containing this Arp2 suppressor identified a structural change that also occurs upon Arp2/3 activation by nucleation promoting factors. These data demonstrate the importance of Arp2 and Arp3 nucleotide binding for nucleating activity, and Arp3 nucleotide binding for maintenance of cortical actin cytoskeleton cytoarchitecture. The Rockefeller University Press 2005-01-17 /pmc/articles/PMC2171590/ /pubmed/15657399 http://dx.doi.org/10.1083/jcb.200408177 Text en Copyright © 2005, The Rockefeller University Press This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).
spellingShingle Research Articles
Martin, Adam C.
Xu, Xiao-Ping
Rouiller, Isabelle
Kaksonen, Marko
Sun, Yidi
Belmont, Lisa
Volkmann, Niels
Hanein, Dorit
Welch, Matthew
Drubin, David G.
Effects of Arp2 and Arp3 nucleotide-binding pocket mutations on Arp2/3 complex function
title Effects of Arp2 and Arp3 nucleotide-binding pocket mutations on Arp2/3 complex function
title_full Effects of Arp2 and Arp3 nucleotide-binding pocket mutations on Arp2/3 complex function
title_fullStr Effects of Arp2 and Arp3 nucleotide-binding pocket mutations on Arp2/3 complex function
title_full_unstemmed Effects of Arp2 and Arp3 nucleotide-binding pocket mutations on Arp2/3 complex function
title_short Effects of Arp2 and Arp3 nucleotide-binding pocket mutations on Arp2/3 complex function
title_sort effects of arp2 and arp3 nucleotide-binding pocket mutations on arp2/3 complex function
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2171590/
https://www.ncbi.nlm.nih.gov/pubmed/15657399
http://dx.doi.org/10.1083/jcb.200408177
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