Sequence analysis of experimentally confirmed SigH promoters.

<p>25 promoters experimentally identified in this research plus 7 previously identified promoters recognized by SigH were analyzed using the MEME analysis suite. <b>A</b>. LOGO showing consensus -10 and -35 promoter elements derived from these sequences. <b>B</b>. 25 promoter sequences that match the SigH consensus sequences. The position of the TSP and the ratio of heat stress-induced expression in wild type relative to expression in a Δ<i>sigH</i> strain are also shown. <b>C</b>. 7 promoter sequences that do not match the SigH consensus.</p

Genes linked to SigH binding sites that show SigH-dependent expression following heat stress.

<p>Genes linked to SigH binding sites that show SigH-dependent expression following heat stress.</p

Expression of <i>sigH</i> and <i>sigH</i>-FLAG in <i>M</i>. <i>tuberculosis</i>.

<p><b>A.</b><i>sigH</i>-FLAG expression was induced in <i>M</i>. <i>tuberculosis</i> H37Rv (wild type) containing a vector expressing <i>sigH</i>-FLAG under control of a TetR-regulated promoter. Samples were obtained at serial time points following addition of aTc to a final concentration of 200 ng/ml, protein was extracted and Western blotting was performed with an anti-FLAG antibody (Sigma-Aldrich). <b>B</b>. <i>M</i>. <i>tuberculosis</i> H37Rv (wild type) or the Δ<i>sigH</i> strain were exposed to 52°C for 15 minutes. The same strains containing a vector expressing <i>sigH</i>-FLAG under control of a TetR-regulated promoter were induced by addition aTc to a final concentration of of 200 ng/ml (aTc). RNA was extracted after 24h and qRT-PCR was performed and analyzed as described in the Materials and Methods. The higher level of induced <i>sigH</i> expression in wild type compared to the Δ<i>sigH</i> strain likely results from increased expression in wild type of the native copy of <i>sigH</i> from its SigH-regulated promoter following induction of the TetR-regulated copy of <i>sigH</i>.</p

ChIP-Seq results for <i>M</i>. <i>tuberculosis</i> SigH.

<p><b>A</b>. Sequencing read coverage for a region with a known SigH binding site 5’ of <i>sigE</i> (Rv1221) in two independent experiments. The total coverage is shown in blue, and the forward and reverse coverages are shown in red and green, respectively. The binding displays the expected shift in position between the forward and reverse reads. <b>B.</b> Genome-wide fold read coverage.</p