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We visualized individual fluorescently labeled Arp2\/3 complexes dynamically interacting with and producing branches on growing actin filaments in vitro. Branch formation was strikingly inefficient, even in the presence of VCA: only \u223c1% of filament-bound Arp2\/3 complexes yielded a daughter filament. VCA acted at multiple steps, increasing both the association rate of Arp2\/3 complexes with mother filament and the fraction of filament-bound complexes that nucleated a daughter. The results lead to a quantitative kinetic mechanism for branched actin assembly, revealing the steps that can be stimulated by additional cellular factors.<\/jats:p>","DOI":"10.1073\/pnas.1211164110","type":"journal-article","created":{"date-parts":[[2013,1,5]],"date-time":"2013-01-05T02:26:17Z","timestamp":1357352777000},"page":"1285-1290","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":116,"title":["Pathway of actin filament branch formation by Arp2\/3 complex revealed by single-molecule imaging"],"prefix":"10.1073","volume":"110","author":[{"given":"Benjamin A.","family":"Smith","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Brandeis University, Waltham, MA 02454-9110; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karen","family":"Daugherty-Clarke","sequence":"additional","affiliation":[{"name":"Department of Biology and Rosenstiel Basic Medical Sciences Research Center, Brandeis University, Waltham, MA 02454-9110"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bruce L.","family":"Goode","sequence":"additional","affiliation":[{"name":"Department of Biology and Rosenstiel Basic Medical Sciences Research Center, Brandeis University, Waltham, MA 02454-9110"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeff","family":"Gelles","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Brandeis University, Waltham, MA 02454-9110; 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