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Journal of Bacteriology, January 2006, p. 150-159, Vol. 188, No. 1
0021-9193/06/$08.00+0     doi:10.1128/JB.188.1.150-159.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

The Mycobacterium tuberculosis TrcR Response Regulator Represses Transcription of the Intracellularly Expressed Rv1057 Gene, Encoding a Seven-Bladed ß-Propeller

Shelley E. Haydel1*,{dagger} and Josephine E. Clark-Curtiss1,2,{dagger}

Departments of Biology,1 Molecular Microbiology, Washington University, St. Louis, Missouri 631302

Received 3 August 2005/ Accepted 11 October 2005

The Mycobacterium tuberculosis TrcR response regulator binds and regulates its own promoter via an AT-rich sequence. Sequences within this AT-rich region determined to be important for TrcR binding were used to search the M. tuberculosis H37Rv genome to identify additional related TrcR binding sites. A similar AT-rich sequence was identified within the intergenic region located upstream of the Rv1057 gene. In the present work, we demonstrate that TrcR binds to a 69-bp AT-rich sequence within the Rv1057 intergenic region and generates specific contacts on the same side of the DNA helix. An M. tuberculosis trcRS deletion mutant, designated STS10, was constructed and used to determine that TrcR functions as a repressor of Rv1057 expression. Additionally, identification of the Rv1057 transcriptional start site suggests that a SigE-regulated promoter also mediates control of Rv1057 expression. Using selective capture of transcribed sequences (SCOTS) analysis as an evaluation of intracellular expression, Rv1057 was shown to be expressed during early M. tuberculosis growth in human macrophages, and the Rv1057 expression profile correlated with a gene that would be repressed by TrcR. Based on structural predictions, motif analyses, and molecular modeling, Rv1057 consists of a series of antiparallel ß-strands which adopt a ß-propeller fold, and it was determined to be the only seven-bladed ß-propeller encoded in the M. tuberculosis genome. These results provide evidence of TrcR response regulator repression of the Rv1057 ß-propeller gene that is expressed during growth of M. tuberculosis within human macrophages.

* Corresponding author. Present address: Center for Infectious Diseases and Vaccinology, The Biodesign Institute, School of Life Sciences, Arizona State University, Tempe, AZ 85287. Phone: (480) 727-7234. Fax: (480) 965-6899. E-mail: Shelley.Haydel{at}asu.edu.

{dagger} Present address: Center for Infectious Diseases and Vaccinology, The Biodesign Institute, School of Life Sciences, Arizona State University, Tempe, AZ 85287.

Journal of Bacteriology, January 2006, p. 150-159, Vol. 188, No. 1
0021-9193/06/$08.00+0     doi:10.1128/JB.188.1.150-159.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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