Expression, autoregulation, and DNA binding properties of the Mycobacterium tuberculosis TrcR response regulator.

Journal of Bacteriology
Shelley E HaydelJ E Clark-Curtiss

Abstract

The TrcRS two-component system of Mycobacterium tuberculosis is comprised of the TrcS histidine kinase and the TrcR response regulator, which is homologous to the OmpR class of DNA binding response regulators. Reverse transcription-PCRs with total RNA showed that the trcR and trcS two-component system genes are transcribed in broth-grown M. tuberculosis. Analysis of the trcR and trcS genes using various SCOTS (selective capture of transcribed sequences) probes also confirmed that these genes are expressed in broth-grown cultures and after 18 h of M. tuberculosis growth in cultured human primary macrophages. To determine if the TrcR response regulator is autoregulated, a trcR-lacZ fusion plasmid and a TrcR expression plasmid were cotransformed into Escherichia coli. Upon induction of the TrcR protein, there was a >500-fold increase in beta-galactosidase activity from the trcR-lacZ fusion, indicating that TrcR is involved in transcriptional autoactivation. Gel mobility shift assays with the trcR promoter and TrcR established that the response regulator was autoregulating via direct binding. By use of a delimiting series of overlapping trcR PCR fragments in gel mobility shift assays with TrcR, an AT-rich region of the trcR promote...Continue Reading

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Citations

Apr 24, 2012·Journal of Microbiological Methods·Kristy L Nowak-LovatoElizabeth Hong-Geller
Jul 4, 2006·Journal of Bacteriology·Yih-Ling TzengDavid S Stephens
Sep 2, 2008·Journal of Bacteriology·Jesús Gonzalo-AsensioCarlos Martín
Dec 1, 2011·Microbiology and Molecular Biology Reviews : MMBR·Daniel J BretlThomas C Zahrt
Oct 20, 2010·BMC Genomics·Marlene Castro-MelchorWei-Shou Hu
Oct 18, 2011·Biochemical and Biophysical Research Communications·Monolekha Bhattacharya, Amit Kumar Das
Apr 1, 2008·Microbiology·Margaret E BauerStanley M Spinola
Jul 5, 2007·Methods in Enzymology·Paul A TuckerJens Preben Morth
Feb 18, 2016·Journal of Bacteriology·Kelly FlentieChristina L Stallings
Feb 1, 2014·Microbiology Spectrum·Tanya Parish

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