title: The Methanosarcina acetivorans thioredoxin system activates DNA binding of the redox-sensitive transcriptional regulator MsvR.
abstract: The production of biogas (methane) by an anaerobic digestion is an important facet to renewable energy, but is subject to instability due to the sensitivity of strictly anaerobic methanogenic archaea (methanogens) to environmental perturbations, such as oxygen. An understanding of the oxidant-sensing mechanisms used by methanogens may lead to the development of more oxidant tolerant (i.e., stable) methanogen strains. MsvR is a redox-sensitive transcriptional regulator that is found exclusively in methanogens. We show here that oxidation of MsvR from Methanosarcina acetivorans (MaMsvR) with hydrogen peroxide oxidizes cysteine thiols, which inactivates MaMsvR binding to its own promoter (P(msvR)). Incubation of oxidized MaMsvR with the M. acetivorans thioredoxin system (NADPH, MaTrxR, and MaTrx7) results in reduction of the cysteines back to thiols and activation of P msvR binding. These data confirm that cysteines are critical for the thiol-disulfide regulation of P(msvR) binding by MaMsvR and support a role for the M. acetivorans thioredoxin system in the in vivo activation of MaMsvR. The results support the feasibility of using MaMsvR and P(msvR), along with the Methanosarcina genetic system, to design methanogen strains with oxidant-regulated gene expression systems, which may aid in stabilizing anaerobic digestion.
authors: Department of Biological Sciences, University of Arkansas-Fayetteville, Fayetteville, AR, 72701, USA.
authors: Ryan
authors: R
authors: Sheehan
authors: Addison C
authors: AC
authors: McCarver
authors: Catherine E
authors: CE
authors: Isom
authors: Elizabeth A
authors: EA
authors: Karr
authors: Daniel J
authors: DJ
authors: Lessner
mesh_terms: D000693:Anaerobiosis; D019843:Archaeal Proteins / Q000378:metabolism*; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D004220:Disulfides / Q000378:metabolism; D019848:Gene Expression Regulation, Archaeal / Q000187:drug effects; D006861:Hydrogen Peroxide / Q000378:metabolism / Q000494:pharmacology; D017020:Methanosarcina / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D009249:NADP / Q000378:metabolism; D016877:Oxidants / Q000378:metabolism / Q000494:pharmacology; D010084:Oxidation-Reduction / Q000187:drug effects; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding / Q000187:drug effects; D013438:Sulfhydryl Compounds / Q000378:metabolism; D013880:Thioredoxin-Disulfide Reductase / Q000378:metabolism; D013879:Thioredoxins / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic / Q000187:drug effects / Q000235:genetics
chemical_list: D019843:Archaeal Proteins; D004220:Disulfides; D016877:Oxidants; D013438:Sulfhydryl Compounds; D014157:Transcription Factors; D013879:Thioredoxins; D009249:NADP; D004247:DNA; D006861:Hydrogen Peroxide; D013880:Thioredoxin-Disulfide Reductase; D003545:Cysteine