Activation of the Arp2/3 Complex by the Actin Filament Binding Protein Abp1p

doi:10.1083/jcb.153.3.627

Published online 30 April 2001. doi:10.1083/jcb.153.3.627

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© The Rockefeller University Press, 0021-9525/2001/4/627/ $5.00
The Journal of Cell Biology, Volume 153, Number 3, April 30, 2001 627-634

Original Article

Activation of the Arp2/3 Complex by the Actin Filament Binding Protein Abp1p

Bruce L. Goode^a, Avital A. Rodal^a, Georjana Barnes^a, and David G. Drubin^a
^a Department of Molecular and Cell Biology, University of California Berkeley, Berkeley, California 94720-3202

Correspondence to: David G. Drubin, Department of Molecular and Cell Biology, University of California Berkeley, 401 Barker Hall, Berkeley, CA 94720-3202. Tel:(510) 642-3692 Fax:(510) 643-0062 E-mail:drubin{at}uclink4.berkeley.edu.

The actin-related protein (Arp) 2/3 complex plays a centralrole in assembly of actin networks. Because distinct actin-basedstructures mediate diverse processes, many proteins are likelyto make spatially and temporally regulated interactions withthe Arp2/3 complex. We have isolated a new activator, Abp1p,which associates tightly with the yeast Arp2/3 complex. Abp1pcontains two acidic sequences (DDW) similar to those found inSCAR/WASp proteins. We demonstrate that mutation of these sequencesabolishes Arp2/3 complex activation in vitro. Genetic studiesindicate that this activity is important for Abp1p functionsin vivo. In contrast to SCAR/WASp proteins, Abp1p binds specificallyto actin filaments, not monomers. Actin filament binding ismediated by the ADF/cofilin homology (ADF-H) domain of Abp1pand is required for Arp2/3 complex activation in vitro. We demonstratethat Abp1p recruits Arp2/3 complex to the sides of filaments,suggesting a novel mechanism of activation. Studies in yeastand mammalian cells indicate that Abp1p is involved functionallyin endocytosis. Based on these results, we speculate that Abp1pmay link Arp2/3-mediated actin assembly to a specific step inendocytosis.

Key Words: actin, yeast, Arp2/3 complex, Abp1, endocytosis

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