PMID:1569108
| Citation |
DiRusso, CC, Heimert, TL and Metzger, AK (1992) Characterization of FadR, a global transcriptional regulator of fatty acid metabolism in Escherichia coli. Interaction with the fadB promoter is prevented by long chain fatty acyl coenzyme A. J. Biol. Chem. 267:8685-91 |
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| Abstract |
The Escherichia coli fadR gene product, FadR, is a multifunctional regulator of fatty acid metabolism. In this work we have purified FadR by a two-step procedure employing two ion-exchange columns. The amino-terminal sequence of the purified protein confirms the sequence of the protein derived from analysis of the DNA sequence (DiRusso, C. C. (1988) Nucleic Acids Res. 16, 7995-8009) and indicates that the initiating methionine is cleaved from the mature protein. Purified FadR binds to a 326-base pair HaeIII fragment of fadB DNA which carries the fadB promoter. DNase I footprinting localizes the operator to a sequence, 5' ATCTGGTACGACCAGAT 3', at +1 to +17 nucleotides relative to the start of transcription. Using protein-DNA gel retention assays, we estimate the Keq of FadR binding to the fadB operator to be approximately 3 x 10(-10) M. Binding of FadR is specifically inhibited by long chain fatty acyl-CoA compounds. The apparent Ki values for oleoyl-CoA, palmitoyl-CoA, and palmitoleoyl-CoA are each 5 nM while that of myristoyl-CoA is 250 nM. Decanoyl-CoA, crotonoyl-CoA, and free fatty acids inhibited binding only at concentrations above 1 microM. |
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| Keywords |
Acyl Coenzyme A/metabolism; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Base Sequence; Chromatography, Liquid; DNA Fingerprinting; DNA, Bacterial/genetics; DNA, Bacterial/metabolism; Electrophoresis, Polyacrylamide Gel; Escherichia coli/genetics; Escherichia coli/metabolism; Fatty Acids/metabolism; Molecular Sequence Data; Operator Regions, Genetic; Plasmids; Promoter Regions, Genetic; Repressor Proteins; Transcription, Genetic |
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