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Applied and Environmental Microbiology, September 2004, p. 5349-5356, Vol. 70, No. 9
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.9.5349-5356.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

RsbT and RsbV Contribute to {sigma}B-Dependent Survival under Environmental, Energy, and Intracellular Stress Conditions in Listeria monocytogenes

Soraya Chaturongakul and Kathryn J. Boor*

Department of Food Science, Cornell University, Ithaca, New York

Received 14 January 2004/ Accepted 17 May 2004

Sigma B ({sigma}B) is a stress-responsive alternative sigma factor that has been identified in various gram-positive bacteria. Seven different regulators of sigma B (Rsbs) are located in the sigB operons of both Bacillus subtilis and Listeria monocytogenes. In B. subtilis, these proteins contribute to regulation of {sigma}B activity by conveying environmental and energy stress signals through two well-established branches of a signal transduction pathway. RsbT contributes to regulation of {sigma}B activity in response to environmental stresses, while RsbV contributes to {sigma}B activation under both environmental and energy stresses in B. subtilis. To probe L. monocytogenes Rsb roles in {sigma}B-mediated responses to various stresses, in-frame deletions were created in rsbT and rsbV. Phenotypic characterization of the L. monocytogenes rsbT and rsbV null mutants revealed that both mutants were similar to the {Delta}sigB strain in their abilities to survive under environmental stress conditions (exposure to synthetic gastric fluid, pH 2.5, acidified brain heart infusion broth [BHI], or oxidative stress [13 mM cumene hydroperoxide]). Under energy stress conditions (carbon starvation in defined media, entry into stationary phase, or reduced intracellular ATP), both {Delta}rsbT and {Delta}rsbV showed survival reductions similar to that of the {Delta}sigB strain. These observations suggest that the pathways for Rsb-dependent regulation of {sigma}B activity differ between L. monocytogenes and B. subtilis. As {sigma}B also activates transcription of the L. monocytogenes prfAP2 promoter, we evaluated virulence-associated characteristics of {Delta}prfAP1rsbT and {Delta}prfAP1rsbV double mutants in hemolysis and tissue culture assays. Both double mutants showed identical phenotypes to {Delta}prfAP1P2 and {Delta}prfAP1sigB double mutants, i.e., reduced hemolysis activity and reduced plaque size in mouse fibroblast cells. These findings indicate that RsbT and RsbV both contribute to {sigma}B activation in L. monocytogenes during exposure to environmental and energy stresses as well as during tissue culture infection.

* Corresponding author. Mailing address: Department of Food Science, 413 Stocking Hall, Cornell University, Ithaca, NY 14853. Phone: (607) 255-3111. Fax: (607) 254-4868. E-mail: kjb4{at}cornell.edu.

Applied and Environmental Microbiology, September 2004, p. 5349-5356, Vol. 70, No. 9
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.9.5349-5356.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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