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Infection and Immunity, July 2001, p. 4276-4286, Vol. 69, No. 7
0019-9567/01/$04.00+0 DOI: 10.1128/IAI.69.7.4276-4286.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Identification of Legionella pneumophila
rcp, a pagP-Like Gene That Confers Resistance to
Cationic Antimicrobial Peptides and Promotes Intracellular
Infection
Marianne
Robey,
William
O'Connell, and
Nicholas P.
Cianciotto*
Department of Microbiology-Immunology,
Northwestern University Medical School, Chicago, Illinois 60611
Received 30 November 2000/Returned for modification 15 January
2001/Accepted 28 March 2001
In the course of characterizing a locus involved in heme
utilization, we identified a Legionella pneumophila gene
predicted to encode a protein with homology to the product of the
Salmonella enterica serovar Typhimurium pagP
gene. In Salmonella, pagP increases resistance to the
bactericidal effects of cationic antimicrobial peptides (CAMPs).
Mutants with insertions in the L. pneumophila pagP-like
gene were generated and showed decreased resistance to different
structural classes of CAMPs compared to the wild type; hence, this gene
was designated rcp for resistance to cationic antimicrobial
peptides. Furthermore, Legionella CAMP resistance was
induced by growth in low-magnesium medium. To determine whether rcp had any role in intracellular survival, mutants were
tested in the two most relevant host cells for Legionnaires' disease, i.e., amoebae and macrophages. These mutants exhibited a
1,000-fold-decreased recovery during a Hartmannella
vermiformis coculture. Complementation of the infectivity defect
could be achieved by introduction of a plasmid containing the intact
rcp gene. Mutations in rcp consistently reduced
both the numbers of bacteria recovered during intracellular infection
and their cytopathic capacity for U937 macrophages. The rcp
mutant was also more defective for lung colonization of A/J mice.
Growth of rcp mutants in buffered yeast extract broth was
identical to that of the wild type, indicating that the observed differences in numbers of bacteria recovered from host cells were not
due to a generalized growth defect. However, in low-Mg2+
medium, the rcp mutant was impaired in stationary-phase
survival. This is the first demonstration of a pagP-like
gene, involved in resistance to CAMPs, being required for intracellular
infection and virulence.
*
Corresponding author. Mailing address: Department of
Microbiology-Immunology, Northwestern University Medical School, 320 East Superior St., Chicago, IL 60611-3010. Phone: (312) 503-1034. Fax:
(312) 503-1339. E-mail: n-cianciotto{at}northwestern.edu.
Infection and Immunity, July 2001, p. 4276-4286, Vol. 69, No. 7
0019-9567/01/$04.00+0 DOI: 10.1128/IAI.69.7.4276-4286.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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