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Journal of Bacteriology, July 1999, p. 4266-4274, Vol. 181, No. 14
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

A Mycobacterial Extracytoplasmic Sigma Factor Involved in Survival following Heat Shock and Oxidative Stress

Norvin D. Fernandes, Qi-long Wu,dagger Dequan Kong,Dagger Xiaoling Puyang, Sumeet Garg, and Robert N. Husson*

Division of Infectious Diseases, Children's Hospital, Harvard Medical School, Boston, Massachusetts 02115

Received 10 September 1998/Accepted 5 May 1999

Extracytoplasmic function (ECF) sigma factors are a heterogeneous group of alternative sigma factors that regulate gene expression in response to a variety of conditions, including stress. We previously characterized a mycobacterial ECF sigma factor, SigE, that contributes to survival following several distinct stresses. A gene encoding a closely related sigma factor, sigH, was cloned from Mycobacterium tuberculosis and Mycobacterium smegmatis. A single copy of this gene is present in these and other fast- and slow-growing mycobacteria, including M. fortuitum and M. avium. While the M. tuberculosis and M. smegmatis sigH genes encode highly similar proteins, there are multiple differences in adjacent genes. The single in vivo transcriptional start site identified in M. smegmatis and one of two identified in M. bovis BCG were found to have -35 promoter sequences that match the ECF-dependent -35 promoter consensus. Expression from these promoters was strongly induced by 50°C heat shock. In comparison to the wild type, an M. smegmatis sigH mutant was found to be more susceptible to cumene hydroperoxide stress but to be similar in logarithmic growth, stationary-phase survival, and survival following several other stresses. Survival of an M. smegmatis sigH sigE double mutant was found to be markedly decreased following 53°C heat shock and following exposure to cumene hydroperoxide. Expression of the second gene in the sigH operon is required for complementation of the sigH stress phenotypes. SigH is an alternative sigma factor that plays a role in the mycobacterial stress response.

* Corresponding author. Mailing address: Division of Infectious Diseases, Children's Hospital, Enders 609, 300 Longwood Ave., Boston, MA 02115. Phone: (617) 355-5151. Fax: (617) 355-8387. E-mail: husson{at}a1.tch.harvard.edu.

dagger Present address: Dyax Corporation, Cambridge, MA 02139.

Dagger Present address: Department of Cardiology, Brigham and Women's Hospital, Boston, MA 02115.

Journal of Bacteriology, July 1999, p. 4266-4274, Vol. 181, No. 14
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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