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Microbiology 152 (2006), 2749-2756; DOI  10.1099/mic.0.28924-0
© 2006 Society for General Microbiology

Regulation of the expression of whiB1 in Mycobacterium tuberculosis: role of cAMP receptor protein

Nisheeth Agarwal, Tirumalai R. Raghunand and William R. Bishai

Department of Medicine, Johns Hopkins University, CRB2, Rm 1.08, 1550 Orleans Street, Baltimore, MD 21231-1044, USA

Correspondence
William R. Bishai
wbishai{at}jhmi.edu

The wbl (whiB-like) genes encode putative transcription factors unique to actinomycetes. This study characterized the promoter element of one of the seven wbl genes of Mycobacterium tuberculosis, whiB1 (Rv3219c). The results reveal that whiB1 is transcribed by a class I-type cAMP receptor protein (CRP)-dependent promoter, harbouring a CRP-binding site positioned at –58.5 with respect to its transcription start point. In vivo promoter activity analysis and electrophoretic mobility shift assays suggest that the expression of whiB1 is indeed regulated by cAMP-dependent binding of CRPM (encoded by the M. tuberculosis gene Rv3676) to the whiB1 5' untranslated region (5'UTR). beta-Galactosidase gene fusion analysis revealed induction of the whiB1 promoter in M. tuberculosis on addition of exogenous dibutyric cAMP (a diffusible cAMP analogue) only when an intact CRP-binding site was present. These results indicate that M. tuberculosis whiB1 transcription is regulated in part by cAMP levels via direct binding of cAMP-activated CRPM to a consensus CRP-binding site in the whiB1 5'UTR.


Abbreviations: CRP, cAMP receptor protein; CRPM, CRP encoded by the M. tuberculosis gene Rv3676; db-cAMP, dibutyric cAMP; EMSA, electrophoretic mobility shift assay; 5'-RACE, rapid amplification of cDNA ends; TSP, transcription start point; 5'UTR, 5' untranslated region




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