Regulation of aroL expression by TyrR protein and Trp repressor in Escherichia coli K-12.
The promoter-operator region of the aroL gene of Escherichia coli K-12 contains three TYR R boxes and one TrpR binding site. Mutational analysis showed that TYR R boxes 1 and 3 are essential for TyrR-mediated regulation of aroL expression, while a fully functional TYR R box 2 does not appear to be essential for regulation. Regulation mediated by the TrpR protein required the TYR R boxes and TrpR site to be functional and was observed in vivo only with a tyrR+ strain. Under conditions favoring the formation of TyrR hexamers, DNase I protection experiments revealed the presence of phased hypersensitive sites, indicative of DNA backbone strain. This suggests that TyrR-mediated repression involves DNA looping. Purified TrpR protein protected the putative TrpR binding site in the presence of tryptophan, and this protection was slightly enhanced in the presence of TyrR protein. This result along with the in vivo findings implies that TyrR and TrpR are able to interact in some way. Inserting 4 bp between TYR R box 1 and the TrpR binding site results in increased tyrosine repression and the abolition of the tryptophan effect. Identification of a potential integration host factor binding site and repression studies of a himA mutant support the notion that integration host factor binding normally exerts a negative effect on tyrosine-mediated repression.
Department of Microbiology, University of Melbourne, Parkville, Victoria, Australia.
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Lawley
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Pittard
D001426:Bacterial Proteins; D001483:Base Sequence; D001665:Binding Sites; D004794:Enzyme Repression; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D005786:Gene Expression Regulation; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D034982:Integration Host Factors; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009875:Operator Regions, Genetic / Q000235:genetics; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000096:biosynthesis*; D011485:Protein Binding; D012097:Repressor Proteins / Q000378:metabolism*; D014443:Tyrosine / Q000494:pharmacology
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D034982:Integration Host Factors; D012097:Repressor Proteins; C027190:TRPR protein, E coli; C069256:TyrR protein, E coli; D014443:Tyrosine; D017853:Phosphotransferases (Alcohol Group Acceptor); C022642:shikimate kinase