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© The Rockefeller University Press, 0022-1007/1997/11/1487/ $5.00
The Journal of Experimental Medicine, Volume 186, Number 9, November 3, 1997 1487-1494

Article

Requirements for Both Rac1 and Cdc42 in Membrane Ruffling and Phagocytosis in Leukocytes

Dianne Cox*, Peter Chang*, Qing Zhang*, P. Gopal Reddy*, Gary M. Bokoch{ddagger}, and Steven Greenberg*

From the * Pulmonary Division, Department of Medicine, Columbia University College of Physicians and Surgeons, New York 10032; and {ddagger} Department of Immunology and Department of Cell Biology, Scripps Research Institute, La Jolla, California 92037

Specific pathways linking heterotrimeric G proteins and Fc{gamma} receptors to the actin-based cytoskeleton are poorly understood. To test a requirement for Rho family members in cytoskeletal events mediated by structurally diverse receptors in leukocytes, we transfected the full-length human chemotactic peptide receptor in RAW 264.7 cells and examined cytoskeletal alterations in response to the chemotactic peptide formyl-methionyl-leucyl-phenylalanine (FMLP), colony stimulating factor–1 (CSF-1), IgG-coated particles, and phorbol 12-myristate 13-acetate (PMA). Expression of Rac1 N17, Cdc42 N17, or the GAP domain of n-chimaerin inhibited cytoskeletal responses to FMLP and CSF-1, and blocked phagocytosis. Accumulation of F-actin– rich "phagocytic cups" was partially inhibited by expression of Rac1 N17 or Cdc42 N17. In contrast, PMA-induced ruffling was not inhibited by expression of Rac1 N17, but was blocked by expression of Cdc42 N17, indicating that cytoskeletal inhibition by these constructs was nonoverlapping. These results demonstrate differential requirements for Rho family GTPases in leukocyte motility, and indicate that both Rac1 and Cdc42 are required for Fc{gamma} receptor– mediated phagocytosis and for membrane ruffling mediated by structurally distinct receptors in macrophages.

Address correspondence to Dr. Steven Greenberg, Columbia University, Department of Medicine, 630 West 168th St., New York, NY 10032. Phone: 212-305-1586; FAX: 212-305-1146; E-mail: greenberg{at}cuccfa.ccc.columbia.edu

1 Abbreviations used in this paper: Chimaerin-GAP, the GAP domain of n-chimaerin; IgG-RBCs, sheep erythrocytes opsonized with rabbit IgG; IPTG, isopropyl β-D-thiogalactopyranoside; RAW cells, RAW 264.7 mouse macrophage cell line.


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