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J Biol Chem, Vol. 275, Issue 1, 141-146, January 7, 2000

Rac1 and Cdc42 Are Required for Phagocytosis, but Not NF-kappa B-dependent Gene Expression, in Macrophages Challenged with Pseudomonas aeruginosa*

Donna J. LeeDagger §, Dianne Cox, Juncheng Li, and Steven Greenbergpar **

From the Departments of Dagger  Pediatrics,  Medicine, and par  Pharmacology, Columbia University, New York, New York 10032

Macrophages respond to Gram-negative bacterial pathogens by phagocytosis and pro-inflammatory gene expression. These responses may require GTPases that have been implicated in cytoskeletal alterations and activation of NF-kappa B. To determine the role of Rac1 and Cdc42 in signal transduction events triggered by Pseudomonas aeruginosa, we expressed GTP binding-deficient alleles of Rac1 or Cdc42, or Chim-GAP, a Rac1/Cdc42-specific GTPase-activating protein domain, in a subline of RAW 264.7 cells, and challenged the transfected cells with a laboratory strain of P. aeruginosa, PAO1. Expression of Rac1 N17, Cdc42 N17, or Chim-GAP led to a marked reduction of phagocytosis. In contrast, nuclear translocation of p65 NF-kappa B was unaffected by expression of the same constructs. Incubation of macrophages with PAO1 led to NF-kappa B-dependent expression of inducible nitric-oxide synthase, COX-2, and tumor necrosis factor-alpha , which was unaffected by inhibition of Rac1 or Cdc42 function. Isogenic strains of PAO1 that lacked surface adhesins were poorly ingested; however, they induced pro-inflammatory gene expression with an efficiency equal to that of PAO1. These results indicate that the signal transduction events leading to phagocytosis and pro-inflammatory protein expression are distinct. Rac1 and Cdc42 serve as effectors of phagocytosis, but not NF-kappa B-dependent gene expression, in the macrophage response to P. aeruginosa.

* This work was supported in part by Grants HL54164 and AI42848 from the National Institutes of Health.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ Supported by the Cystic Fibrosis Foundation.

** Established Investigator of the American Heart Association. To whom correspondence should be addressed: Depts. of Medicine and Pharmacology/PH8C, Columbia University, 630 W. 168th St., New York, NY 10032. Tel.: 212-305-1586; Fax: 212-305-1146; E-mail: greenberg@cuccfa.ccc.columbia.edu.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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