title: Activation of Arp2/3 complex: addition of the first subunit of the new filament by a WASP protein triggers rapid ATP hydrolysis on Arp2.
abstract: In response to activation by WASP-family proteins, the Arp2/3 complex nucleates new actin filaments from the sides of preexisting filaments. The Arp2/3-activating (VCA) region of WASP-family proteins binds both the Arp2/3 complex and an actin monomer and the Arp2 and Arp3 subunits of the Arp2/3 complex bind ATP. We show that Arp2 hydrolyzes ATP rapidly-with no detectable lag-upon nucleation of a new actin filament. Filamentous actin and VCA together do not stimulate ATP hydrolysis on the Arp2/3 complex, nor do monomeric and filamentous actin in the absence of VCA. Actin monomers bound to the marine macrolide Latrunculin B do not polymerize, but in the presence of phalloidin-stabilized actin filaments and VCA, they stimulate rapid ATP hydrolysis on Arp2. These data suggest that ATP hydrolysis on the Arp2/3 complex is stimulated by interaction with a single actin monomer and that the interaction is coordinated by VCA. We show that capping of filament pointed ends by the Arp2/3 complex (which occurs even in the absence of VCA) also stimulates rapid ATP hydrolysis on Arp2, identifying the actin monomer that stimulates ATP hydrolysis as the first monomer at the pointed end of the daughter filament. We conclude that WASP-family VCA domains activate the Arp2/3 complex by driving its interaction with a single conventional actin monomer to form an Arp2-Arp3-actin nucleus. This actin monomer becomes the first monomer of the new daughter filament.
authors.affiliation: Graduate Group in Biophysics, University of California, San Francisco, USA.
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Dayel
authors.forename: R Dyche
authors.initials: RD
authors.lastname: Mullins
mesh_terms: D000048:Acanthamoeba / Q000378:metabolism; D051377:Actin-Related Protein 2 / Q000737:chemistry*; D051378:Actin-Related Protein 3 / Q000737:chemistry*; D000199:Actins / Q000737:chemistry; D000251:Adenosine Triphosphatases / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000737:chemistry*; D000818:Animals; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D002467:Cell Nucleus / Q000378:metabolism; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D008856:Microscopy, Fluorescence; D010710:Phosphates / Q000737:chemistry; D011108:Polymers / Q000737:chemistry; D011485:Protein Binding; D013844:Thiazoles / Q000494:pharmacology; D053778:Thiazolidines; D051316:Wiskott-Aldrich Syndrome Protein / Q000737:chemistry*
chemical_list: D051377:Actin-Related Protein 2; D051378:Actin-Related Protein 3; D000199:Actins; D019086:Bridged Bicyclo Compounds, Heterocyclic; D010710:Phosphates; D011108:Polymers; D013844:Thiazoles; D053778:Thiazolidines; D051316:Wiskott-Aldrich Syndrome Protein; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; C037068:latrunculin B