title: Role for CysJ flavin reductase in molybdenum cofactor-dependent resistance of Escherichia coli to 6-N-hydroxylaminopurine.
abstract: We have previously described a novel Escherichia coli detoxification system for the removal of toxic and mutagenic N-hydroxylated nucleobases and related compounds that requires the molybdenum cofactor. Two subpathways (ycbX and yiiM) were identified, each employing a novel molybdo activity capable of inactivating N-hydroxylated compounds by reduction to the corresponding amine. In the present study, we identify the cysJ gene product as one additional component of this system. While the CysJ protein has been identified as the NADPH:flavin oxidoreductase component of the CysJI sulfite reductase complex (CysJ(8)I(4)), we show that the role of CysJ in base analog detoxification is unique and independent of CysI and sulfite reductase. We further show that CysJ functions as a specific partner of the YcbX molybdoenzyme. We postulate that the function of CysJ in this pathway is to provide, via its NADPH:flavin reductase activity, the reducing equivalents needed for the detoxification reaction at the YcbX molybdocenter. In support of the proposed interaction of the CysJ and YcbX proteins, we show that an apparent CysJ-YcbX "hybrid" protein from two Vibrio species is capable of compensating for a double cysJ ycbX defect in E. coli.
authors.affiliation: Laboratory of Molecular Genetics, MD E3-01, National Institute of Environmental Health Sciences, 111 TW Alexander Drive, Research Triangle Park, NC 27709, USA.
authors.forename: Stanislav G
authors.initials: SG
authors.lastname: Kozmin
authors.forename: Jian
authors.initials: J
authors.lastname: Wang
authors.forename: Roel M
authors.initials: RM
authors.lastname: Schaaper
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives* / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology; D003067:Coenzymes / Q000378:metabolism*; D003545:Cysteine / Q000494:pharmacology; D024881:Drug Resistance, Bacterial / Q000235:genetics / Q000502:physiology*; D004926:Escherichia coli / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000502:physiology*; D038181:FMN Reductase / Q000235:genetics / Q000502:physiology*; D008667:Metalloproteins / Q000378:metabolism*; D000090006:Molybdenum Cofactors; D016366:Open Reading Frames / Q000235:genetics; D009876:Operon / Q000235:genetics; D011621:Pteridines / Q000378:metabolism*
chemical_list: D000225:Adenine; D001426:Bacterial Proteins; D003067:Coenzymes; D003545:Cysteine; D029968:Escherichia coli Proteins; D038181:FMN Reductase; D008667:Metalloproteins; D000090006:Molybdenum Cofactors; D011621:Pteridines; C032610:6-N-hydroxylaminopurine; C022261:molybdopterin; C540768:Fre protein, E coli