title: Redox-sensitive DNA binding by homodimeric Methanosarcina acetivorans MsvR is modulated by cysteine residues.
abstract: BACKGROUND
  Methanoarchaea are among the strictest known anaerobes, yet they can survive exposure to oxygen. The mechanisms by which they sense and respond to oxidizing conditions are unknown. MsvR is a transcription regulatory protein unique to the methanoarchaea. Initially identified and characterized in the methanogen Methanothermobacter thermautotrophicus (Mth), MthMsvR displays differential DNA binding under either oxidizing or reducing conditions. Since MthMsvR regulates a potential oxidative stress operon in M. thermautotrophicus, it was hypothesized that the MsvR family of proteins were redox-sensitive transcription regulators.
  
  
  RESULTS
  An MsvR homologue from the methanogen Methanosarcina acetivorans, MaMsvR, was overexpressed and purified. The two MsvR proteins bound the same DNA sequence motif found upstream of all known MsvR encoding genes, but unlike MthMsvR, MaMsvR did not bind the promoters of select genes involved in the oxidative stress response. Unlike MthMsvR that bound DNA under both non-reducing and reducing conditions, MaMsvR bound DNA only under reducing conditions. MaMsvR appeared as a dimer in gel filtration chromatography analysis and site-directed mutagenesis suggested that conserved cysteine residues within the V4R domain were involved in conformational rearrangements that impact DNA binding.
  
  
  CONCLUSIONS
  Results presented herein suggest that homodimeric MaMsvR acts as a transcriptional repressor by binding Ma PmsvR under non-reducing conditions. Changing redox conditions promote conformational changes that abrogate binding to Ma PmsvR which likely leads to de-repression.
authors.affiliation: Department of Microbiology and Plant Biology, University of Oklahoma, 770 Van Vleet Oval, Norman, OK, 73019, USA.
authors.forename: Catherine E
authors.initials: CE
authors.lastname: Isom
authors.forename: Jessica L
authors.initials: JL
authors.lastname: Turner
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Lessner
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Karr
mesh_terms: D019843:Archaeal Proteins / Q000302:isolation & purification / Q000378:metabolism*; D001665:Binding Sites; D003545:Cysteine / Q000235:genetics / Q000378:metabolism*; D004252:DNA Mutational Analysis; D019641:DNA, Archaeal / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D017013:Methanobacteriaceae / Q000378:metabolism; D017020:Methanosarcina / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D010084:Oxidation-Reduction; D011401:Promoter Regions, Genetic; D055503:Protein Multimerization; D012097:Repressor Proteins / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D019843:Archaeal Proteins; D019641:DNA, Archaeal; D004268:DNA-Binding Proteins; D012097:Repressor Proteins; D003545:Cysteine