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Journal of Bacteriology, July 1999, p. 3890-3897, Vol. 181, No. 13
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Effect of rpoS Mutation on the Stress
Response and Expression of Virulence Factors in Pseudomonas
aeruginosa
Sang-Jin
Suh,1,2,*
Laura
Silo-Suh,2
Donald E.
Woods,3
Daniel J.
Hassett,4
Susan E. H.
West,5 and
Dennis E.
Ohman2
Department of Microbiology and Immunology,
University of Tennessee, and Veterans Affairs Medical Center, Memphis,
Tennessee 381041; Department of
Microbiology and Immunology, Medical College of Virginia campus of
Virginia Commonwealth University, and McGuire Veterans Affairs Medical
Center, Richmond, Virginia 23298-06782;
Department of Microbiology and Infectious Diseases, University
of Calgary, Calgary, Alberta, Canada T2N 4N13;
Department of Molecular Genetics, Biochemistry and
Microbiology, University of Cincinnati College of Medicine, Cincinnati,
Ohio 452574; and Department of
Pathobiological Sciences, School of Veterinary Medicine, University
of Wisconsin
Madison, Madison, Wisconsin 537065
Received 7 December 1998/Accepted 20 April 1999
The sigma factor RpoS (
S) has been described as a
general stress response regulator that controls the expression of genes which confer increased resistance to various stresses in some gram-negative bacteria. To elucidate the role of RpoS in
Pseudomonas aeruginosa physiology and pathogenesis, we
constructed rpoS mutants in several strains of P. aeruginosa, including PAO1. The PAO1 rpoS mutant was
subjected to various environmental stresses, and we compared the
resistance phenotype of the mutant to that of the parent. The PAO1
rpoS mutant was slightly more sensitive to carbon
starvation than the wild-type strain, but this phenotype was obvious
only when the cells were grown in a medium supplemented with glucose as
the sole carbon source. In addition, the PAO1 rpoS mutant
was hypersensitive to heat shock at 50°C, increased osmolarity, and
prolonged exposure to high concentrations of
H2O2. In accordance with the hypersensitivity
to H2O2, catalase production was 60% lower in
the rpoS mutant than in the parent strain. We also assessed
the role of RpoS in the production of several exoproducts known to be
important for virulence of P. aeruginosa. The
rpoS mutant produced 50% less exotoxin A, but it produced
only slightly smaller amounts of elastase and LasA protease than the
parent strain. The levels of phospholipase C and casein-degrading
proteases were unaffected by a mutation in rpoS in PAO1.
The rpoS mutation resulted in the increased production of
the phenazine antibiotic pyocyanin and the siderophore pyoverdine. This
increased pyocyanin production may be responsible for the enhanced
virulence of the PAO1 rpoS mutant that was observed in a
rat chronic-lung-infection model. In addition, the rpoS
mutant displayed an altered twitching-motility phenotype, suggesting
that the colonization factors, type IV fimbriae, were affected.
Finally, in an alginate-overproducing cystic fibrosis (CF) isolate,
FRD1, the rpoS101::aacCI mutation
almost completely abolished the production of alginate when the
bacterium was grown in a liquid medium. On a solid medium, the FRD1
rpoS mutant produced approximately 70% less alginate than
did the wild-type strain. Thus, our data indicate that although some of
the functions of RpoS in P. aeruginosa physiology are
similar to RpoS functions in other gram-negative bacteria, it also has
some functions unique to this bacterium.
*
Corresponding author. Mailing address: Department of
Microbiology and Immunology, Medical College of Virginia campus of
Virginia Commonwealth University, 1101 East Marshall St., P.O. Box
980678, Richmond, VA 23298-0678. Phone: (804) 628-0247. Fax: (804)
828-9946. E-mail: ssuh{at}hsc.vcu.edu.
Journal of Bacteriology, July 1999, p. 3890-3897, Vol. 181, No. 13
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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