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Applied and Environmental Microbiology, August 2005, p. 4241-4247, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4241-4247.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Role for HtrA in Stress Induction and Virulence Potential in Listeria monocytogenes

Helena M. Stack,1 Roy D. Sleator,1 Megan Bowers,2 Colin Hill,1* and Cormac G. M. Gahan1

Department of Microbiology and Alimentary Pharmabiotic Centre, University College Cork, Cork, Ireland,1 New York University School of Medicine, New York, New York2

Received 29 December 2004/ Accepted 2 February 2005

In silico analysis of the Listeria monocytogenes genome revealed lmo0292, a gene predicted to encode a HtrA-like serine protease. A stable insertion mutant was constructed, revealing a requirement for htrA in the listerial response to heat, acid, and penicillin stress. Transcriptional analysis revealed that htrA is not induced in response to heat shock but is induced in response to low pH and penicillin G stress. Furthermore, htrA expression was shown to be dependent upon the LisRK two-component sensor-kinase, a system known to respond to changes in integrity of the cell envelope. In addition, we demonstrated that a second in-frame start codon, upstream of that previously annotated for L. monocytogenes htrA, incorporating a putative signal sequence appears to influence virulence potential. Finally, a significant virulence defect was observed for the htrA mutant, indicating that this gene is required for full virulence in mice. Our findings suggest that L. monocytogenes lmo0292 encodes an HtrA-like serine protease that is not part of the classical heat shock response but is involved in stress responses and virulence.

* Corresponding author. Mailing address: Department of Microbiology, University College Cork, Cork, Ireland. Phone: (353) 21-4901373. Fax: (353) 21-4903101. E-mail: c.hill{at}ucc.ie.

Applied and Environmental Microbiology, August 2005, p. 4241-4247, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4241-4247.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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