Rac1 and Cdc42 are required for phagocytosis, but not NF-κB-dependent gene expression, in macrophages challenged with Pseudomonas aeruginosa

Donna J. Lee, Dianne Cox, Juncheng Li, Steven Greenberg

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

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- κB. 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-κB was unaffected by expression of the same constructs. Incubation of macrophages with PAO1 led to NF-κB-dependent expression of inducible nitric-oxide synthase, COX-2, and tumor necrosis factor-α, 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-κB-dependent gene expression, in the macrophage response to P. aeruginosa.

Original languageEnglish (US)
Pages (from-to)141-146
Number of pages6
JournalJournal of Biological Chemistry
Volume275
Issue number1
DOIs
StatePublished - Jan 7 2000
Externally publishedYes

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Macrophages
Phagocytosis
Gene expression
Pseudomonas aeruginosa
Signal transduction
Gene Expression
GTPase-Activating Proteins
Signal Transduction
GTP Phosphohydrolases
Nitric Oxide Synthase Type II
Pathogens
Guanosine Triphosphate
Tumor Necrosis Factor-alpha
Chemical activation
Alleles
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Rac1 and Cdc42 are required for phagocytosis, but not NF-κB-dependent gene expression, in macrophages challenged with Pseudomonas aeruginosa. / Lee, Donna J.; Cox, Dianne; Li, Juncheng; Greenberg, Steven.

In: Journal of Biological Chemistry, Vol. 275, No. 1, 07.01.2000, p. 141-146.

Research output: Contribution to journalArticle

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