Journal
JOURNAL OF CELL BIOLOGY
Volume 179, Issue 2, Pages 239-245Publisher
ROCKEFELLER UNIV PRESS
DOI: 10.1083/jcb.200705122
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Actin assembly at the leading edge of migrating cells depends on the availability of high- affinity free barbed ends ( FBE) that drive actin. lament elongation and subsequent membrane protrusion. We investigated the specific mechanisms through which the Rac1 and Rac2 small guanosine triphosphatases (GTPases) generate free barbed ends in neutrophils. Using neutrophils lacking either Rac1 or Rac2 and a neutrophil permeabilization model that maintains receptor signaling to the actin cytoskeleton, we assessed the mechanisms through which these two small GTPases mediate FBE generation downstream of the formyl- methionyl- leucylphenylalanine receptor. We demonstrate here that uncapping of existing barbed ends is mediated through Rac1, whereas cofilin- and ARP2/3-mediated FBE generation are regulated through Rac2. This unique combination of experimental tools has allowed us to identify the relative roles of uncapping (15%), cofilin severing (10%), and ARP2/ 3 de novo nucleation (75%) in FBE generation and the respective roles played by Rac1 and Rac2 in mediating actin dynamics.
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