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© The Rockefeller University Press,
0021-9525/2000//785 $5.00
The Journal of Cell Biology, Volume 150, Number 4,
, 2000 785-796
Original Article |
Two Pathways through Cdc42 Couple the N-Formyl Receptor to Actin Nucleation in Permeabilized Human Neutrophils
mglogauer{at}rics.bwh.harvard.edu
We developed a permeabilization method that retains coupling between N-formyl-methionyl-leucyl-phenylalanine tripeptide (FMLP) receptor stimulation, shape changes, and barbed-end actin nucleation in human neutrophils. Using GTP analogues, phosphoinositides, a phosphoinositide-binding peptide, constitutively active or inactive Rho GTPase mutants, and activating or inhibitory peptides derived from neural Wiskott-Aldrich syndrome family proteins (N-WASP), we identified signaling pathways leading from the FMLP receptor to actin nucleation that require Cdc42, but then diverge. One branch traverses the actin nucleation pathway involving N-WASP and the Arp2/3 complex, whereas the other operates through active Rac to promote actin nucleation. Both pathways depend on phosphoinositide expression. Since maximal inhibition of the Arp2/3 pathway leaves an N17Rac inhibitable alternate pathway intact, we conclude that this alternate involves phosphoinositide-mediated uncapping of actin filament barbed ends.
Key Words: Arp2/3 actin assembly signal transduction pathways Rac FMLP
© 2000 The Rockefeller University Press
Abbreviations used in this paper: F-actin, filamentous actin; FMLP, N-formyl methionyl leucyl phenylalanine tripeptide; GST, glutathione S-transferase; N-WASP, neural Wiskott-Aldrich syndrome family proteins; OG, n-octyl-β-glucopyranoside; PIP2, phosphatidylinositol bisphosphate.
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