Genomic Studies with Escherichia coli MelR Protein: Applications of Chromatin Immunoprecipitation and Microarrays
- 15 October 2004
- journal article
- research article
- Published by American Society for Microbiology in Journal of Bacteriology
- Vol. 186 (20) , 6938-6943
- https://doi.org/10.1128/jb.186.20.6938-6943.2004
Abstract
Escherichia coli MelR protein is a transcription activator that is essential for melibiose-dependent expression of the melAB genes. We have used chromatin immunoprecipitation to study the binding of MelR and RNA polymerase to the melAB promoter in vivo. Our results show that MelR is associated with promoter DNA, both in the absence and presence of the inducer melibiose. In contrast, RNA polymerase is recruited to the melAB promoter only in the presence of inducer. The MelR DK261 positive control mutant binds to the melAB promoter but cannot recruit RNA polymerase. Further analysis of immunoprecipitated DNA, by using an Affymetrix GeneChip array, showed that the melAB promoter is the major, if not the sole, target in E. coli for MelR. This was confirmed by a transcriptomics experiment to analyze RNA in cells either with or without melR.Keywords
This publication has 15 references indexed in Scilit:
- Regulators Encoded in the Escherichia coli Type III Secretion System 2 Gene Cluster Influence Expression of Genes within the Locus for Enterocyte Effacement in Enterohemorrhagic E. coli O157:H7Infection and Immunity, 2004
- Transcription activation at the Escherichia coli melAB promoter: interactions of MelR with its DNA target site and with domain 4 of the RNA polymerase σ subunitMolecular Microbiology, 2004
- Identifying global regulators in transcriptional regulatory networks in bacteriaCurrent Opinion in Microbiology, 2003
- DNA binding of the transcription activator protein MelR from Escherichia coli and its C-terminal domainNucleic Acids Research, 2002
- Evidence for “Pre-recruitment” as a New Mechanism of Transcription Activation in Escherichia coli: The Large Excess of SoxS Binding Sites per Cell Relative to the Number of SoxS Molecules per CellBiochemical and Biophysical Research Communications, 2002
- In vivo assays to examine transcription factor localization and target gene specificityMethods, 2002
- Complex formation between activator and RNA polymerase as the basis for transcriptional activation by MarA and SoxS in Escherichia coliMolecular Microbiology, 2002
- The Complete Genome Sequence of Escherichia coli K-12Science, 1997
- Organisation of the regulatory region of the Escherichia coli melibiose operonGene, 1987
- A plasmid vector that allows fusion of the Escherichia coli galactokinase gene to the translation startpoint of other genesFEBS Letters, 1985