A protein that binds to the regulatory region of the Escherichia coli ilvIH operon.
The ilvIH operon of Escherichia coli encodes acetohydroxyacid synthase III, an isoenzyme involved in branched-chain amino acid biosynthesis. Transcription of the ilvIH operon is repressed by growing cells in the presence of leucine (C.H. Squires, M. DeFelice, S.R. Wessler, and J.M. Calvo, J. Bacteriol. 147:797-804, 1981). A protein in crude extracts of E. coli, termed the ilvIH-binding (IHB) protein, bound specifically in vitro to DNA upstream of the ilvIH operon. The binding protein, partially purified by Polymin precipitation, gel filtration, and phosphocellulose chromatography, has a native molecular weight of 43,000 and is composed of two subunits of identical size. As determined by protection against lambda exonuclease and DNase I, the protein binds within a region -190 to -260 relative to the start point of transcription. In addition, the IHB protein binds to a site between positions -100 and -40. The following evidence suggests that binding of this protein to the region upstream of ilvIH is related to the regulation of this operon by leucine. Binding of the IHB protein to the ilvIH regulatory region in vitro was reduced by leucine but not by isoleucine, valine, or threonine. In a mutant strain isolated by M.V. Ursini, P. Arcari, and M. DeFelice (Mol. Gen. Genet. 181:491-496, 1981), transcription was not repressed by leucine. A protein in extracts of this mutant strain bound to the ilvIH regulatory region, but the complex migrated through agarose gels with a mobility different from that of the complex formed by wild-type protein. Furthermore, a concentration of leucine that substantially reduced binding of the wild-type to DNA did not affect binding of the protein from the mutant strain. A simple model consistent with these findings is that transcription from the ilvIH promoter is stimulated by binding the IHB protein to one or more sites upstream of the promoter and that leucine interferes with this binding.
Ricca
E
E
Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, New York 14853.
Aker
D A
DA
Calvo
J M
JM
D000095:Acetolactate Synthase / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005796:Genes; D005798:Genes, Bacterial; D005809:Genes, Regulator; D007527:Isoenzymes / Q000235:genetics*; D009876:Operon; D007652:Oxo-Acid-Lyases / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping
D004268:DNA-Binding Proteins; D007527:Isoenzymes; D000095:Acetolactate Synthase; D007652:Oxo-Acid-Lyases