Figure 3

MsvR binding and regulatory targets assessed by EMSA. (a) Sequences of the 50 bp region of Ma P _msvR used to confirm MaMsvR binding to boxes A and B. Sequence changes within the binding boxes are shown. (b) EMSA assays with the template (50 nM) variations shown in (a) and 1 μM (20-fold excess over DNA) reduced MaMsvR (R, 5 mM DTT). A 50 bp region of Ma P _msvR was included as a binding control. The gel wells are indicated (W). (c) EMSA analysis with reduced MaMsvR (R, 5 mM DTT) and its own promoter (Ma P _msvR , 10 nM), various intergenic regions of an oxidative stress response cluster (Ma P ₄₆₆₄ , P ₃₇₃₄ , P ₃₇₃₆ , 10 nM) as well as the control Ma histone A promoter (Ma P _hmaA , 10 nM). A region of rpoK (10 nM) was tested for binding because an MsvR binding site (TTCGN₈CGAA) is present in the coding region. The gel wells are indicated (W).