Dual control by regulatory gene fdsR of the fds operon encoding the NAD+-linked formate dehydrogenase of Ralstonia eutropha.
The transcriptional regulator gene fdsR was identified 150 bp upstream of the divergently oriented fdsGBACD operon encoding the soluble, NAD+-linked formate dehydrogenase in the chemoautotrophic bacterium Ralstonia eutropha H16. Its deduced product, FdsR, displays a basal sequence similarity to the regulatory proteins of the LysR family. The carboxy-terminal domain of FdsR contains a short region that is conserved in formate dehydrogenases. Deletion of fdsR revealed a dual regulatory effect of FdsR on the fds operon by acting as transcriptional activator in the presence of formate or as repressor in the absence of formate. Studies with fdsR transcriptional fusions also suggested a negative autoregulation of the gene. A promoter structure resembling sigma70-dependent promoters from Escherichia coli was identified upstream of the fdsR transcriptional start site. FdsR purified to homogeneity after overexpression of fdsR in E. coli is a 130 kDa homotetramer binding to the fds control region located between the fdsR and fdsG genes. Formate significantly increased the binding affinity of FdsR for this region. Two FdsR binding sites characterized by the inverted-repeat structure ATANG-N10-CNTAT were identified. The regulatory pattern found in R. eutropha was also observed in the heterologous host E. coli and results from a novel mode of control of formate dehydrogenase genes.
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J I
JI
Institut für Mikrobiologie und Genetik, Georg-August-Universität Göttingen, Grisebachstrasse 8, D-37077 Göttingen, Germany.
Bowien
B
B
D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D001483:Base Sequence; D020118:Cupriavidus necator / Q000201:enzymology / Q000235:genetics*; D005560:Formate Dehydrogenases / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D005809:Genes, Regulator; D008969:Molecular Sequence Data; D009243:NAD / Q000378:metabolism; D009876:Operon; D010957:Plasmids / Q000235:genetics; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D017422:Sequence Analysis, DNA; D015534:Trans-Activators / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
D001426:Bacterial Proteins; C402927:FdsR protein, Ralstonia eutropha; D012097:Repressor Proteins; D015534:Trans-Activators; D009243:NAD; D005560:Formate Dehydrogenases