Expression of a Butyrivibrio fibrisolvens E14 gene (cinB) encoding an enzyme with cinnamoyl ester hydrolase activity is negatively regulated by the product of an adjacent gene (cinR).
A second cinnamoyl ester hydrolase (CEH) encoding gene (cinB) has been characterized from the ruminal bacterium Butyrivibrio fibrisolvens E14. CinB is more similar to CinA (previously named CinI) (28% amino acid identify), the first CEH described from B. fibrisolvens E14, than either of the enzymes are to any other member of the family of hydrolases to which they belong. Upstream of cinB, and in the opposite orientation, is a gene (cinR) encoding a protein with substantial similarity to members of the MarR family of negative regulators of bacterial gene expression. By alignment of these sequences, a possible helix-turn-helix DNA-binding domain has been identified. CinR was expressed at a high level in Escherichia coli using the lac promoter. In E. coli CinR repressed the expression of CinB, but had no effect on the expression of CinA. In gel mobility-shift assays, CinR bound specifically to the cinR-cinB intergenic region. Two identical 16 nucleotide inverted repeats adjacent to the putative PcinR and PcinB promoters are likely binding sites for CinR. The addition of FAXX (O-[5-O-(trans-feruloyl)-alpha-L-arabinofuranosyl]-(1,3)- O-beta-D-xylopyranosyl-(1,4)-D-xylopyranose) and Fara [5-O-(trans-feruloyl)-arabinofuranose], but not xylobiose, ferulic acid and a number of other soluble components of hemicellulose, inhibited the binding of CinR to DNA.
Dalrymple
Brian P
BP
Division of Tropical Animal production. Commonwealth Scientific and Industrial Research Organisation, Private Bag no. 3 PO, Indooroopilly, Qld 4068, Australia.
Swadling
Yolande
Y
Division of Tropical Animal production. Commonwealth Scientific and Industrial Research Organisation, Private Bag no. 3 PO, Indooroopilly, Qld 4068, Australia.
D000595:Amino Acid Sequence; D001089:Arabinose / Q000031:analogs & derivatives / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics; D001438:Bacteroidaceae / Q000235:genetics*; D001483:Base Sequence; D002265:Carboxylic Ester Hydrolases / Q000096:biosynthesis / Q000235:genetics*; D003001:Cloning, Molecular; D003373:Coumaric Acids / Q000378:metabolism; D004252:DNA Mutational Analysis; D004268:DNA-Binding Proteins / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D005809:Genes, Regulator; D008957:Models, Genetic; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D011994:Recombinant Proteins / Q000096:biosynthesis; D012097:Repressor Proteins / Q000235:genetics; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D014312:Trisaccharides / Q000378:metabolism
D001426:Bacterial Proteins; D003373:Coumaric Acids; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C080004:MarR protein, E coli; D011994:Recombinant Proteins; D012097:Repressor Proteins; D014312:Trisaccharides; C064239:O-(5-O-(feruloyl)-alpha-arabinofuranosyl)-(1-3)-O-beta-xylopyranosyl-(1-4)-xylopyranose; C072516:5-O-feruloylarabinose; D001089:Arabinose; D002265:Carboxylic Ester Hydrolases; C070196:feruloyl esterase