title: Novel regulatory system nemRA-gloA for electrophile reduction in Escherichia coli K-12.
abstract: Electrophilic compounds such as glyoxals, which are toxic due to their reactive carbonyl group, are generated in vivo through various pathways. In this study, we obtained evidence indicating that the nemRA operon, previously reported to encode a repressor and the N-ethylmaleimide reductase, respectively, is co-transcribed with the 3'-proximal gloA gene encoding glyoxalase I. The operon is not only involved in cytosolic detoxification but is also regulated by electrophiles such as quinones and glyoxals. A gel mobility shift assay revealed that purified NemR repressor bound to DNA was dissociated upon interaction with quinones and glyoxals, while their reduced forms were ineffective. The cysteines of NemR at 21 and 116 were essential in sensing electrophiles in vivo and in vitro. Reversible intermolecular disulphide bonds were observed with a reducing agent as well as with electrophiles. DNA binding affinity reduced by glyoxal was also increased with a reducing agent. The NemA reductase, an FMN-containing enzyme, exhibited catalytic activity toward various electrophiles including quinones, while GloA played a major role in glyoxal detoxification. Therefore, we propose that cells have a cytosolic system consisting of the nemRA-gloA operon for the reduction of electrophiles, especially quinones and glyoxals, to maintain an appropriate intracellular redox balance.
authors.affiliation: Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Yuseong-gu, Daejeon 305-701, Korea.
authors.forename: Changhan
authors.initials: C
authors.lastname: Lee
authors.forename: Jongcheol
authors.initials: J
authors.lastname: Shin
authors.forename: Chankyu
authors.initials: C
authors.lastname: Park
mesh_terms: D003600:Cytosol / Q000201:enzymology; D048168:Escherichia coli K12 / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D006037:Glyoxal / Q000378:metabolism* / Q000494:pharmacology; D007791:Lactoylglutathione Lyase / Q000235:genetics / Q000378:metabolism*; D009876:Operon; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D011809:Quinones / Q000378:metabolism* / Q000494:pharmacology; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D029968:Escherichia coli Proteins; C531319:NemR protein, E coli; D011809:Quinones; D014157:Transcription Factors; D006037:Glyoxal; D010088:Oxidoreductases; C040723:alpha,beta-ketoalkene reductase; D007791:Lactoylglutathione Lyase