The long term objective of this proposal is to gain a better understanding of how Mycobacterium tuberculosis establishes infection so that effective strategies can be developed to prevent it. A multidisciplinary approach will focus on the interaction of tubercle bacilli with macrophages at the molecular, genetic and cellular levels, with an emphasis on M. tuberculosis gene expression within macrophages.
The specific aims are: 1) Identifying M. tuberculosis genes that are induced when bacteria are within macrophages by 2D gel electrophoresis coupled with mass spectrometry. 2) Identifying class-specific regulatory motifs among intracellular induced promoters using a combination of molecular and computational techniques. 3) Characterizing the roles in the M. tuberculosis-macrophage interaction of selected intracellular induced M. tuberculosis genes. Assays will include bacterial survival, replication and trafficking in macrophages. 4) Assessing the role of cAMP signaling in M. tuberculosis within macrophages by defining the distribution and expression among mycobacteria of genes encoding novel cyclase and cyclic NMP binding proteins; estimating the minimum number of cAMP-responsive proteins using 2D gels; and determining the effects of a novel adenylate cyclase gene knock-out on M. tuberculosis interaction with macrophages using tissue culture, microscopy, and 2D gel analyses. This work will contribute to our understanding of the factors needed for the establishment of tuberculosis infection and disease and will identify potential targets for tuberculosis vaccines, therapeutics, and diagnostic purposes.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI045658-05
Application #
6746933
Study Section
Bacteriology and Mycology Subcommittee 2 (BM)
Program Officer
Sizemore, Christine F
Project Start
2000-07-01
Project End
2006-08-31
Budget Start
2004-06-01
Budget End
2006-08-31
Support Year
5
Fiscal Year
2004
Total Cost
$299,357
Indirect Cost
Name
Wadsworth Center
Department
Type
DUNS #
153695478
City
Menands
State
NY
Country
United States
Zip Code
12204
Girardin, Roxie C; Bai, Guangchun; He, Jie et al. (2018) AbmR (Rv1265) is a novel transcription factor of Mycobacterium tuberculosis that regulates host cell association and expression of the non-coding small RNA Mcr11. Mol Microbiol 110:811-830
Ranganathan, Sridevi; Bai, Guangchun; Lyubetskaya, Anna et al. (2016) Characterization of a cAMP responsive transcription factor, Cmr (Rv1675c), in TB complex mycobacteria reveals overlap with the DosR (DevR) dormancy regulon. Nucleic Acids Res 44:134-51
Bai, Guangchun; Knapp, Gwendowlyn S; McDonough, Kathleen A (2011) Cyclic AMP signalling in mycobacteria: redirecting the conversation with a common currency. Cell Microbiol 13:349-58
Bai, Guangchun; Schaak, Damen D; McDonough, Kathleen A (2009) cAMP levels within Mycobacterium tuberculosis and Mycobacterium bovis BCG increase upon infection of macrophages. FEMS Immunol Med Microbiol 55:68-73
Vasudeva-Rao, Hema M; McDonough, Kathleen A (2008) Expression of the Mycobacterium tuberculosis acr-coregulated genes from the DevR (DosR) regulon is controlled by multiple levels of regulation. Infect Immun 76:2478-89
Bai, Guangchun; Gazdik, Michaela A; Schaak, Damen D et al. (2007) The Mycobacterium bovis BCG cyclic AMP receptor-like protein is a functional DNA binding protein in vitro and in vivo, but its activity differs from that of its M. tuberculosis ortholog, Rv3676. Infect Immun 75:5509-17
Bai, Guangchun; McCue, Lee Ann; McDonough, Kathleen A (2005) Characterization of Mycobacterium tuberculosis Rv3676 (CRPMt), a cyclic AMP receptor protein-like DNA binding protein. J Bacteriol 187:7795-804
Florczyk, Matthew A; McCue, Lee Ann; Purkayastha, Anjan et al. (2003) A family of acr-coregulated Mycobacterium tuberculosis genes shares a common DNA motif and requires Rv3133c (dosR or devR) for expression. Infect Immun 71:5332-43
Purkayastha, Anjan; McCue, Lee Ann; McDonough, Kathleen A (2002) Identification of a Mycobacterium tuberculosis putative classical nitroreductase gene whose expression is coregulated with that of the acr aene within macrophages, in standing versus shaking cultures, and under low oxygen conditions. Infect Immun 70:1518-29
Florczyk, M A; McCue, L A; Stack, R F et al. (2001) Identification and characterization of mycobacterial proteins differentially expressed under standing and shaking culture conditions, including Rv2623 from a novel class of putative ATP-binding proteins. Infect Immun 69:5777-85