Mechanisms for activating the actin-related protein 2/3 (Arp2/3) complex have been the focus of many recent studies. Here, we identify a novel mode of Arp2/3 complex regulation mediated by the highly conserved actin binding protein coronin. Yeast coronin (Crn1) physically associates with the Arp2/3 complex and inhibits WA- and Abp1-activated actin nucleation in vitro. The inhibition occurs specifically in the absence of preformed actin filaments, suggesting that Crn1 may restrict Arp2/3 complex activity to the sides of filaments. The inhibitory activity of Crn1 resides in its coiled coil domain. Localization of Crn1 to actin patches in vivo and association of Crn1 with the Arp2/3 complex also require its coiled coil domain. Genetic studies provide in vivo evidence for these interactions and activities. Overexpression of CRN1 causes growth arrest and redistribution of Arp2 and Crn1p into aberrant actin loops. These defects are suppressed by deletion of the Crn1 coiled coil domain and by arc35-26, an allele of the p35 subunit of the Arp2/3 complex. Further in vivo evidence that coronin regulates the Arp2/3 complex comes from the observation that crn1 and arp2 mutants display an allele-specific synthetic interaction. This work identifies a new form of regulation of the Arp2/3 complex and an important cellular function for coronin.
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23 December 2002
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December 23 2002
Direct regulation of Arp2/3 complex activity and function by the actin binding protein coronin
Christine L. Humphries,
Christine L. Humphries
1Department of Molecular and Medical Genetics, University of Toronto, Ontario M5S 1A8, Canada
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Heath I. Balcer,
Heath I. Balcer
2Biology Department and Rosenstiel Center, Brandeis University, Waltham, MA 02454
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Jessica L. D'Agostino,
Jessica L. D'Agostino
2Biology Department and Rosenstiel Center, Brandeis University, Waltham, MA 02454
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Barbara Winsor,
Barbara Winsor
3Centre National de Recherche Scientifique, Université Louis Pasteur, Strasbourg, France
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David G. Drubin,
David G. Drubin
4Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720
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Georjana Barnes,
Georjana Barnes
4Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720
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Brenda J. Andrews,
Brenda J. Andrews
1Department of Molecular and Medical Genetics, University of Toronto, Ontario M5S 1A8, Canada
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Bruce L. Goode
Bruce L. Goode
2Biology Department and Rosenstiel Center, Brandeis University, Waltham, MA 02454
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Christine L. Humphries
1Department of Molecular and Medical Genetics, University of Toronto, Ontario M5S 1A8, Canada
Heath I. Balcer
2Biology Department and Rosenstiel Center, Brandeis University, Waltham, MA 02454
Jessica L. D'Agostino
2Biology Department and Rosenstiel Center, Brandeis University, Waltham, MA 02454
Barbara Winsor
3Centre National de Recherche Scientifique, Université Louis Pasteur, Strasbourg, France
David G. Drubin
4Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720
Georjana Barnes
4Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720
Brenda J. Andrews
1Department of Molecular and Medical Genetics, University of Toronto, Ontario M5S 1A8, Canada
Bruce L. Goode
2Biology Department and Rosenstiel Center, Brandeis University, Waltham, MA 02454
Address correspondence to Bruce Goode, Rosenstiel Center, Brandeis University, 415 South Street, Waltham, MA 02454. Tel.: (781) 736-2451. Fax: (781) 736-2405. E-mail: [email protected]
C.L. Humphries and H.I. Balcer contributed equally to this work.
*
Abbreviation used in this paper: Arp2/3, actin-related protein 2/3.
Received:
June 26 2002
Revision Received:
September 30 2002
Accepted:
November 07 2002
Online ISSN: 1540-8140
Print ISSN: 0021-9525
The Rockefeller University Press
2002
J Cell Biol (2002) 159 (6): 993–1004.
Article history
Received:
June 26 2002
Revision Received:
September 30 2002
Accepted:
November 07 2002
Citation
Christine L. Humphries, Heath I. Balcer, Jessica L. D'Agostino, Barbara Winsor, David G. Drubin, Georjana Barnes, Brenda J. Andrews, Bruce L. Goode; Direct regulation of Arp2/3 complex activity and function by the actin binding protein coronin . J Cell Biol 23 December 2002; 159 (6): 993–1004. doi: https://doi.org/10.1083/jcb.200206113
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