This research seeks to investigate in greater depth the function of two alternative sigma factors of M. tuberculosis, SigL and SigM, which have been the focus of recent study in the Pi'slaboratory. Data from the Pi's laboratory indicate that these sigma factors regulate genes involved in surface lipid and secreted protein synthesis. These two sigma factors appear to play distinct roles in regulating different aspects of surface lipid biosynthesis in M. tuberculosis. SigM negatively regulates several lipid biosynthetic pathways that have been shown to be important for early stages of M. tuberculosis infection. In contrast, SigL, which is critical for the virulence of this organism, positively regulates two polyketide synthasegenes, pks7 and pkslO, that also have been implicated in virulence. In addition to their role in regulating lipid biosynthesis, both of these sigma factors regulate genes encoding secreted proteins. SigL regulates the gene for the major secreted protein antigen Mpt53, while SigM is required for the expression of two pairs of ESAT-6-like secreted proteins. The hypothesis underlying this research is that these sigma factors and the genes they regulate play key roles in the modulation of the interaction between M. tuberculosis and the host during the course of infection, via regulation of genes for surface lipid and secreted protein production. The overall goal of this proposal is thus to investigate the function of these two sigma factors by characterizing the regulation and function of these lipids and secreted proteins, and their role in host-pathogen interactions. This goal will be achieved through four specific aims.
Aim 1 will characterize the role of SigL in regulating specific lipid biosynthesis, and the role of these lipids in virulence.
Aim 2 will determine the function of the SigL-regulated major secreted protein antigen Mpt53 and its role in virulence.
Aim 3 will characterize the role of SigM in regulating secreted protein and lipid biosynthesis.
Aim 4 will investigate the role of SigM in M. tuberculosis pathogenesis and characterize the cellular immune response to the SigM-regulated secreted proteins EsxE, EsxF, EsxT and EsxU. Health Significance: Tuberculosis remains a major human pathogen worldwide and in the United States, and treatment requires prolonged multi-drug therapy. The insights achieved by this research into M. tuberculosis pathogenesis and the host responses to this pathogen may identify new approaches to developing more effective treatment for this infection.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI037901-14
Application #
7995207
Study Section
Host Interactions with Bacterial Pathogens Study Section (HIBP)
Program Officer
Lacourciere, Karen A
Project Start
1995-06-01
Project End
2012-11-30
Budget Start
2010-12-01
Budget End
2012-11-30
Support Year
14
Fiscal Year
2011
Total Cost
$408,383
Indirect Cost
Name
Children's Hospital Boston
Department
Type
DUNS #
076593722
City
Boston
State
MA
Country
United States
Zip Code
02115
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Ymele-Leki, Patrick; Cao, Shugeng; Sharp, Jared et al. (2012) A high-throughput screen identifies a new natural product with broad-spectrum antibacterial activity. PLoS One 7:e31307
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Song, Taeksun; Song, Seung-Eun; Raman, Sahadevan et al. (2008) Critical role of a single position in the -35 element for promoter recognition by Mycobacterium tuberculosis SigE and SigH. J Bacteriol 190:2227-30
Williams, Diana L; Pittman, Tana L; Deshotel, Mike et al. (2007) Molecular basis of the defective heat stress response in Mycobacterium leprae. J Bacteriol 189:8818-27
Raman, Sahadevan; Puyang, Xiaoling; Cheng, Tan-Yun et al. (2006) Mycobacterium tuberculosis SigM positively regulates Esx secreted protein and nonribosomal peptide synthetase genes and down regulates virulence-associated surface lipid synthesis. J Bacteriol 188:8460-8
Hahn, Mi-Young; Raman, Sahadevan; Anaya, Mauricio et al. (2005) The Mycobacterium tuberculosis extracytoplasmic-function sigma factor SigL regulates polyketide synthases and secreted or membrane proteins and is required for virulence. J Bacteriol 187:7062-71
Raman, Sahadevan; Hazra, Rohan; Dascher, Christopher C et al. (2004) Transcription regulation by the Mycobacterium tuberculosis alternative sigma factor SigD and its role in virulence. J Bacteriol 186:6605-16
Blangero, John (2004) Localization and identification of human quantitative trait loci: king harvest has surely come. Curr Opin Genet Dev 14:233-40

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