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.
Tuberculosis remains a leading killer worldwide, and new approaches for its treatment and prevention are urgently needed. This effort will benefit greatly from a better understanding of gene regulation in Mycobacterium tuberculosis, particularly with respect to this pathogen's response to its host environment. We examined the behavior of two promoters from the divergently transcribed M. tuberculosis genes acr/hspX/Rv2031c (alpha-crystallin homolog) and Rv2032/acg (acr-coregulated gene) by using a promoter-GFP fusion assay in Mycobacterium bovis BCG. We found that Rv2032 is a novel macrophage-induced gene whose expression is coregulated with that of acr. Relative levels of intracellular induction for both promoters were significantly affected by shallow standing versus shaking bacterial culture conditions prior to macrophage infection, and both promoters were strongly induced under low oxygen conditions. Deletion analyses showed that DNA sequences within a 43-bp region were required for expression of these promoters under all conditions. Multiple sequence alignment and database searches performed with PROBE indicated that Rv2032 is one of eight M. tuberculosis genes of previously unknown function that belong to an unusual superfamily of classical nitroreductases, which may have a role for bacteria within the host environment. These findings show that mycobacterial culture conditions can greatly influence the results and interpretation of subsequent gene regulation experiments. We propose that these differences might be exploited for dissection of the regulatory factors that affect mycobacterial gene expression within the host.
Purkayastha
Anjan
A
Department of Biomedical Sciences, University of Albany School of Public Health, Albany, New York 12201-2002, USA.
McCue
Lee Ann
LA
McDonough
Kathleen A
KA
D020816:Amino Acid Motifs; D000818:Animals; D000942:Antigens, Bacterial / Q000096:biosynthesis / Q000235:genetics*; D001431:Bacteriological Techniques; D001483:Base Sequence; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D008264:Macrophages / Q000382:microbiology*; D051379:Mice; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000201:enzymology / Q000235:genetics*; D009601:Nitroreductases / Q000096:biosynthesis / Q000235:genetics*; D010100:Oxygen / Q000378:metabolism; D011401:Promoter Regions, Genetic
D000942:Antigens, Bacterial; C072681:Mycobacterium tuberculosis antigens; D009601:Nitroreductases; D010100:Oxygen