The redox-sensing regulator YodB senses quinones and diamide via a thiol-disulfide switch in Bacillus subtilis.
The MarR/DUF24-type repressor YodB controls the azoreductase AzoR1, the nitroreductase YodC and the redox-sensing regulator Spx in response to quinones and diamide in Bacillus subtilis. Previously, we showed using a yodBCys6-Ala mutant that the conserved Cys6 apparently contributes to the DNA-binding activity of YodB in vivo. Here, we present data that mutation of Cys6 to Ser led to a form of the protein that was reduced in redox-sensing in response to diamide and 2-methylhydroquinone (MHQ) in vivo. DNA-binding experiments indicate that YodB is regulated by a reversible thiol-modification in response to diamide and MHQ in vitro. Redox-regulation of YodB involves Cys6-Cys101' intermolecular disulfide formation by diamide and quinones in vitro. Diagonal Western blot analyses confirm the formation of intersubunit disulfides in YodB in vivo that require the conserved Cys6 and either of the C-terminal Cys101' or Cys108' residues. This study reveals a thiol-disulfide switch model of redox-regulation for the YodB repressor to sense electrophilic compounds in vivo.
Chi
Bui Khanh
BK
Institute for Microbiology, Ernst-Moritz-Arndt-University of Greifswald, Greifswald, Germany.
Albrecht
Dirk
D
Gronau
Katrin
K
Becher
Dörte
D
Hecker
Michael
M
Antelmann
Haike
H
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D003545:Cysteine / Q000378:metabolism; D003958:Diamide / Q000378:metabolism*; D004220:Disulfides / Q000378:metabolism*; D008958:Models, Molecular; D058977:Molecular Sequence Annotation; D008969:Molecular Sequence Data; D009154:Mutation; D010084:Oxidation-Reduction; D011809:Quinones / Q000378:metabolism*; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015203:Reproducibility of Results; D016415:Sequence Alignment; D013438:Sulfhydryl Compounds / Q000378:metabolism
D001426:Bacterial Proteins; D004220:Disulfides; D011809:Quinones; D012097:Repressor Proteins; D013438:Sulfhydryl Compounds; D003958:Diamide; D003545:Cysteine