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Infection and Immunity, May 2000, p. 2493-2502, Vol. 68, No. 5
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

RpmA Is Required for Nonopsonic Phagocytosis of Pseudomonas aeruginosa

David A. Simpson1,* and David P. Speert1,2

Division of Infectious and Immunological Diseases, Department of Pediatrics, and the Canadian Bacterial Diseases Network,1 and Department of Microbiology and Immunology,2 University of British Columbia, Vancouver, British Columbia, Canada V5Z 4H4

Received 6 December 1999/Returned for modification 7 January 2000/Accepted 19 January 2000

Pseudomonas aeruginosa causes severe respiratory tract infections in patients with cystic fibrosis (CF). We have been examining nonopsonic phagocytosis of P. aeruginosa by macrophages. To study the P. aeruginosa-macrophage interaction at the molecular level, we have constructed a transposon Tn5G bank in a clinical isolate of P. aeruginosa (strain 4020) and identified mutants resistant to nonopsonic phagocytosis. Phagocytosis-resistant mutants were enriched by passaging the transposon bank over 18 macrophage monolayers. Of 900 individual mutants isolated from this enriched pool in a nonopsonic phagocytosis assay, we identified 85 putative mutants that were resistant to phagocytosis. In this study, we have characterized one of these transposon mutants, P. aeruginosa 4020 H27A, which was poorly ingested. H27A possessed a Tn5G insertion in a gene encoding a protein with homology to the MotA proteins of several species of bacteria. We have called this gene rpmA for required for phagocytosis by macrophages. RpmA is one of two MotA paralogs in P. aeruginosa. This rpmA::Tn5G mutant was motile both on agar plates and in visual examination of wet mounts. The phagocytosis defect was partially complemented by providing the rpmA gene in trans and fully complemented when both rpmA and rpmB were provided. A rpmA null mutant was ingested by macrophages similar to the H27A transposon mutant. These data suggest that the rpmA and rpmB gene products are required for the efficient ingestion of P. aeruginosa by macrophages.

* Corresponding author. Mailing address: B.C. Research Institute for Children's and Women's Health, 950 W 28th Ave., Rm. 381, Vancouver, BC, Canada V5Z 4H4. Phone: (604) 875-2469. Fax: (604) 875-2226. E-mail: dsimpson{at}cbdn.ca.

Infection and Immunity, May 2000, p. 2493-2502, Vol. 68, No. 5
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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