The Escherichia coli RutR transcription factor binds at targets within genes as well as intergenic regions.
The Escherichia coli RutR protein is the master regulator of genes involved in pyrimidine catabolism. Here we have used chromatin immunoprecipitation in combination with DNA microarrays to measure the binding of RutR across the chromosome of exponentially growing E. coli cells. Twenty RutR-binding targets were identified and analysis of these targets generated a DNA consensus logo for RutR binding. Complementary in vitro binding assays showed high-affinity RutR binding to 16 of the 20 targets, with the four low-affinity RutR targets lacking predicted key binding determinants. Surprisingly, most of the DNA targets for RutR are located within coding segments of the genome and appear to have little or no effect on transcript levels in the conditions tested. This contrasts sharply with other E. coli transcription factors whose binding sites are primarily located in intergenic regions. We suggest that either RutR has yet undiscovered function or that evolution has been slow to eliminate non-functional DNA sites for RutR because they do not have an adverse effect on cell fitness.
Department of Frontier Bioscience and Micro-Nano Technology Research Centre, Hosei University, Koganei, Tokyo 184-8584, Nippon Institute for Biological Science, Ome, Tokyo 198-0024, Japan.
Tomohiro
T
Shimada
Akira
A
Ishihama
Stephen J W
SJ
Busby
David C
DC
Grainger
D001483:Base Sequence; D001665:Binding Sites; D047369:Chromatin Immunoprecipitation; D016384:Consensus Sequence; D004269:DNA, Bacterial / Q000378:metabolism; D021901:DNA, Intergenic / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development; D029968:Escherichia coli Proteins / Q000378:metabolism*; D005798:Genes, Bacterial; D016680:Genome, Bacterial; D020411:Oligonucleotide Array Sequence Analysis; D017422:Sequence Analysis, DNA; D014157:Transcription Factors / Q000378:metabolism*
D004269:DNA, Bacterial; D021901:DNA, Intergenic; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C530619:RutR protein, E coli; D014157:Transcription Factors