Transcription Regulation by the Mycobacterium tuberculosis Alternative Sigma Factor SigD and Its Role in Virulence

ABSTRACT Mycobacterium tuberculosis, an obligate mammalian pathogen, adapts to its host during the course of infection via the regulation of gene expression. Of the regulators of transcription that play a role in this response, several alternative sigma factors of M. tuberculosis have been shown to control gene expression in response to stresses, and some of these are required for virulence or persistence in vivo. For this study, we examined the role of the alternative sigma factor SigD in M. tuberculosis gene expression and virulence. Using microarray analysis, we identified several genes whose expression was altered in a strain with a sigD deletion. A small number of these genes, including sigD itself, the gene encoding the autocrine growth factor RpfC, and a gene of unknown function, Rv1815, appear to be directly regulated by this sigma factor. By identifying the in vivo promoters of these genes, we have determined a consensus promoter sequence that is putatively recognized by SigD. The expression of several genes encoding PE-PGRS proteins, part of a large family of related genes of unknown function, was significantly increased in the sigD mutant. We found that the expression of sigD is stable throughout log phase and stationary phase but that it declines rapidly with oxygen depletion. In a mouse infection model, the sigD mutant strain was attenuated, with differences in survival and the inflammatory response in the lung between mice infected with the mutant and those infected with the wild type.

[1]  R. Fleischmann,et al.  Attenuation of Late-Stage Disease in Mice Infected bythe Mycobacterium tuberculosis Mutant Lacking theSigF Alternate Sigma Factor and Identification ofSigF-Dependent Genes by MicroarrayAnalysis , 2004, Infection and Immunity.

[2]  W. Jacobs,et al.  Individual Mycobacterium tuberculosis Resuscitation-Promoting Factor Homologues Are Dispensable for Growth In Vitro and In Vivo , 2004, Infection and Immunity.

[3]  Dirk Schnappinger,et al.  Inhibition of Respiration by Nitric Oxide Induces a Mycobacterium tuberculosis Dormancy Program , 2003, The Journal of experimental medicine.

[4]  Yang Liu,et al.  Transcriptional Adaptation of Mycobacterium tuberculosis within Macrophages , 2003, The Journal of experimental medicine.

[5]  Harvey Rubin,et al.  The role of RelMtb-mediated adaptation to stationary phase in long-term persistence of Mycobacterium tuberculosis in mice , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[6]  E. Rubin,et al.  Genes required for mycobacterial growth defined by high density mutagenesis , 2003, Molecular microbiology.

[7]  D. Kell,et al.  A family of autocrine growth factors in Mycobacterium tuberculosis , 2002, Molecular microbiology.

[8]  G. Schoolnik,et al.  Role of the extracytoplasmic‐function σ Factor σH in Mycobacterium tuberculosis global gene expression , 2002 .

[9]  R. Fleischmann,et al.  Reduced immunopathology and mortality despite tissue persistence in a Mycobacterium tuberculosis mutant lacking alternative σ factor, SigH , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[10]  Sayera Banu,et al.  Are the PE‐PGRS proteins of Mycobacterium tuberculosis variable surface antigens? , 2002, Molecular microbiology.

[11]  J. Betts,et al.  Evaluation of a nutrient starvation model of Mycobacterium tuberculosis persistence by gene and protein expression profiling , 2002, Molecular microbiology.

[12]  William R. Jacobs,et al.  Evidence that Mycobacterial PE_PGRS Proteins Are Cell Surface Constituents That Influence Interactions with Other Cells , 2001, Infection and Immunity.

[13]  W. Jacobs,et al.  The Alternative Sigma Factor SigH Regulates Major Components of Oxidative and Heat Stress Responses in Mycobacterium tuberculosis , 2001, Journal of bacteriology.

[14]  A. Coates,et al.  Increased levels of sigJ mRNA in late stationary phase cultures of Mycobacterium tuberculosis detected by DNA array hybridisation. , 2001, FEMS microbiology letters.

[15]  G. Schoolnik,et al.  The Mycobacterium tuberculosis ECF sigma factor σE: role in global gene expression and survival in macrophages † , 2001, Molecular microbiology.

[16]  Dirk Schnappinger,et al.  Regulation of the Mycobacterium tuberculosis hypoxic response gene encoding α-crystallin , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[17]  S. Laal,et al.  Antigens of Mycobacterium tuberculosis Expressed during Preclinical Tuberculosis: Serological Immunodominance of Proteins with Repetitive Amino Acid Sequences , 2001, Infection and Immunity.

[18]  W. Bishai,et al.  Construction and Characterization of aMycobacterium tuberculosis Mutant Lacking the Alternate Sigma Factor Gene, sigF , 2000, Infection and Immunity.

[19]  James C. Sacchettini,et al.  Persistence of Mycobacterium tuberculosis in macrophages and mice requires the glyoxylate shunt enzyme isocitrate lyase , 2000, Nature.

[20]  N. Federspiel,et al.  Granuloma-specific expression of Mycobacterium virulence proteins from the glycine-rich PE-PGRS family. , 2000, Science.

[21]  T. Weisbrod,et al.  Characterization of the Mycobacterium tuberculosis iniBAC Promoter, a Promoter That Responds to Cell Wall Biosynthesis Inhibition , 2000, Journal of bacteriology.

[22]  R. Husson,et al.  A Mycobacterial Extracytoplasmic Sigma Factor Involved in Survival following Heat Shock and Oxidative Stress , 1999, Journal of bacteriology.

[23]  R. Husson,et al.  A Mycobacterial Extracytoplasmic Sigma Factor Involved in Survival following Heat Shock and Oxidative Stress , 1999 .

[24]  F. Kramer,et al.  Differential expression of 10 sigma factor genes in Mycobacterium tuberculosis , 1999, Molecular microbiology.

[25]  B. Barrell,et al.  Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence , 1998, Nature.

[26]  W. Jacobs,et al.  Efficient allelic exchange and transposon mutagenesis in Mycobacterium tuberculosis. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[27]  R. Husson,et al.  A mycobacterial extracytoplasmic function sigma factor involved in survival following stress , 1997, Journal of bacteriology.

[28]  L. Wayne,et al.  An in vitro model for sequential study of shiftdown of Mycobacterium tuberculosis through two stages of nonreplicating persistence , 1996, Infection and immunity.

[29]  W. Bishai,et al.  A stationary-phase stress-response sigma factor from Mycobacterium tuberculosis. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[30]  S. Dhandayuthapani,et al.  Gene expression in mycobacteria: transcriptional fusions based on xylE and analysis of the promoter region of the response regulator mtrA from Mycobacterium tuberculosis , 1994, Molecular microbiology.

[31]  K. Rudd,et al.  Analysis of the Streptomyces coelicolor sigE gene reveals the existence of a subfamily of eubacterial RNA polymerase sigma factors involved in the regulation of extracytoplasmic functions. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[32]  W. Jacobs,et al.  Site-specific integration of mycobacteriophage L5: integration-proficient vectors for Mycobacterium smegmatis, Mycobacterium tuberculosis, and bacille Calmette-Guérin. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[33]  W. Jacobs,et al.  Isolation and characterization of efficient plasmid transformation mutants of Mycobacterium smegmatis , 1990, Molecular microbiology.

[34]  D. Sherman,et al.  Regulation of the Mycobacterium tuberculosis hypoxic response gene encoding alpha -crystallin. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[35]  Gary A. Churchill,et al.  Analysis of Variance for Gene Expression Microarray Data , 2000, J. Comput. Biol..