Transcriptional activation of the aldehyde reductase YqhD by YqhC and its implication in glyoxal metabolism of Escherichia coli K-12.
The reactive alpha-oxoaldehydes such as glyoxal (GO) and methylglyoxal (MG) are generated in vivo from sugars through oxidative stress. GO and MG are believed to be removed from cells by glutathione-dependent glyoxalases and other aldehyde reductases. We isolated a number of GO-resistant (GO(r)) mutants from Escherichia coli strain MG1655 on LB plates containing 10 mM GO. By tagging the mutations with the transposon TnphoA-132 and determining their cotransductional linkages, we were able to identify a locus to which most of the GO(r) mutations were mapped. DNA sequencing of the locus revealed that it contains the yqhC gene, which is predicted to encode an AraC-type transcriptional regulator of unknown function. The GO(r) mutations we identified result in missense changes in yqhC and were concentrated in the predicted regulatory domain of the protein, thereby constitutively activating the product of the adjacent gene yqhD. The transcriptional activation of yqhD by wild-type YqhC and its mutant forms was established by an assay with a beta-galactosidase reporter fusion, as well as with real-time quantitative reverse transcription-PCR. We demonstrated that YqhC binds to the promoter region of yqhD and that this binding is abolished by a mutation in the potential target site, which is similar to the consensus sequence of its homolog SoxS. YqhD facilitates the removal of GO through its NADPH-dependent enzymatic reduction activity by converting it to ethadiol via glycolaldehyde, as detected by nuclear magnetic resonance, as well as by spectroscopic measurements. Therefore, we propose that YqhC is a transcriptional activator of YqhD, which acts as an aldehyde reductase with specificity for certain aldehydes, including GO.
Lee
Changhan
C
Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Yuseong-gu, Daejeon, Republic of Korea.
Kim
Insook
I
Lee
Junghoon
J
Lee
Kang-Lok
KL
Min
Bumchan
B
Park
Chankyu
C
D000079:Acetaldehyde / Q000031:analogs & derivatives / Q000378:metabolism; D000449:Aldehyde Reductase / Q000096:biosynthesis*; D000595:Amino Acid Sequence; D019677:Artificial Gene Fusion; D001665:Binding Sites / Q000235:genetics; D004252:DNA Mutational Analysis; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D024881:Drug Resistance, Bacterial; D048168:Escherichia coli K12 / Q000187:drug effects / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000096:biosynthesis* / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter; D008040:Genetic Linkage; D006037:Glyoxal / Q000378:metabolism* / Q000633:toxicity; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D009249:NADP / Q000378:metabolism; D011485:Protein Binding; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D015534:Trans-Activators / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*; D015533:Transcriptional Activation; D014161:Transduction, Genetic; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D015534:Trans-Activators; D014157:Transcription Factors; C551650:YqhC protein, E coli; D006037:Glyoxal; D009249:NADP; D000449:Aldehyde Reductase; C528732:YqhD protein, E coli; D001616:beta-Galactosidase; D000079:Acetaldehyde; C010972:glycolaldehyde