Figure 2From: Redox-sensitive DNA binding by homodimeric Methanosarcina acetivorans MsvR is modulated by cysteine residuesEMSA of MsvR homologues on their respective promoters. The gel wells are indicated (W). (a and b) EMSA to test binding of MaMsvR (a) and MthMsvR (b) to the MaMsvR promoter (Ma P msvR , 10 nM), the MthMsvR/fpaA intergenic promoter region (Mth P msvR/fpaA , 10 nM), and the Mth histone B promoter (Mth P hmtB , 10 nM). Each promoter has a control lane (-) that contains no protein, a binding reaction that contains either Ma or Mth MsvR (200 nM) in the absence of DTT (non-reduced, +), and a binding reaction that contains either Ma or Mth MsvR (200 nM) in the presence of 5 mM DTT (reduced, R). (c) EMSA assay (10 nM Ma P msvR DNA) with decreasing concentrations of reduced MaMsvR (5 mM DTT) [monomer] 1 μM, 500 nM, 250 nM, 125 nM, 62.5 nM, 31.3 nM, 15.6 nM, 7.8 nM, and 3.9 nM. (d) EMSA assay (10 nM Mth P msvR/fpaA DNA) with decreasing concentrations of reduced MaMsvR (5 mM DTT) [monomer] 1 μM, 500 nM, 250 nM, 125 nM, 62.5 nM, 31.3 nM, 15.6 nM, 7.8 nM, and 3.9 nM. (e) EMSA assay (10 nM Mth P msvR/fpaA DNA) with decreasing concentrations of reduced MthMsvR (5 mM DTT) [monomer] 1 μM, 500 nM, 250 nM, 125 nM, 62.5 nM, 31.3 nM, 15.6 nM, 7.8 nM, and 3.9 nM.Back to article page