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Applied and Environmental Microbiology, May 2007, p. 2919-2930, Vol. 73, No. 9
0099-2240/07/$08.00+0     doi:10.1128/AEM.02664-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

The Alternative Sigma Factor {sigma}B and the Virulence Gene Regulator PrfA Both Regulate Transcription of Listeria monocytogenes Internalins{triangledown} ,{dagger}

Patrick McGann, Martin Wiedmann, and Kathryn J. Boor*

Department of Food Science, Cornell University, Ithaca, New York 14853

Received 14 November 2006/ Accepted 21 February 2007

Some Listeria monocytogenes internalins are recognized as contributing to invasion of mammalian tissue culture cells. While PrfA is well established as a positive regulator of L. monocytogenes virulence gene expression, the stress-responsive {sigma}B has been recognized only recently as contributing to expression of virulence genes, including some that encode internalins. To measure the relative contributions of PrfA and {sigma}B to internalin gene transcription, we used reverse transcription-PCR to quantify transcript levels for eight internalin genes (inlA, inlB, inlC, inlC2, inlD, inlE, inlF, and inlG) in L. monocytogenes 10403S and in isogenic {Delta}prfA, {Delta}sigB, and {Delta}sigB {Delta}prfA strains. Strains were grown under defined conditions to produce (i) active PrfA, (ii) active {sigma}B and active PrfA, (iii) inactive PrfA, and (iv) active {sigma}B and inactive PrfA. Under the conditions tested, {sigma}B and PrfA contributed differentially to the expression of the various internalins such that (i) both {sigma}B and PrfA contributed to inlA and inlB transcription, (ii) only PrfA contributed to inlC transcription, (iii) only {sigma}B contributed to inlC2 and inlD transcription, and (iv) neither {sigma}B nor PrfA contributed to inlF and inlG transcription. inlE transcript levels were undetectable. The important role for {sigma}B in regulating expression of L. monocytogenes internalins suggests that exposure of this organism to environmental stress conditions, such as those encountered in the gastrointestinal tract, may activate internalin transcription. Interplay between {sigma}B and PrfA also appears to be critical for regulating transcription of some virulence genes, including inlA, inlB, and prfA.


* Corresponding author. Mailing address: Department of Food Science, Cornell University, Ithaca, NY 14853. Phone: (607) 254-4967. Fax: (607) 254-4868. E-mail: kjb4{at}cornell.edu

{triangledown} Published ahead of print on 2 March 2007.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.


Applied and Environmental Microbiology, May 2007, p. 2919-2930, Vol. 73, No. 9
0099-2240/07/$08.00+0     doi:10.1128/AEM.02664-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.




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