title: The tryptophan repressor sequence is highly conserved among the Enterobacteriaceae.
abstract: Tryptophan biosynthesis in Escherichia coli is regulated by the product of the trpR gene, the tryptophan (Trp) repressor. Trp aporepressor binds the corepressor, L-tryptophan, to form a holorepressor complex, which binds trp operator DNA tightly, and inhibits transcription of the tryptophan biosynthetic operon. The conservation of trp operator sequences among enteric Gram-negative bacteria suggests that trpR genes from other bacterial species can be cloned by complementation in E. coli. To clone trpR homologues, a deletion of the E. coli trpR gene, delta trpR504, was made on a plasmid by site-directed mutagenesis, then crossed onto the E. coli genome. Plasmid clones of the trpR genes of Enterobacter aerogenes and Enterobacter cloacae were isolated by complementation of the delta trpR504 allele, scored as the ability to repress beta-galactosidase synthesis from a prophage-borne trpE-lacZ gene fusion. The predicted amino acid sequences of four enteric TrpR proteins show differences, clustered on the backside of the folded repressor, opposite the DNA-binding helix-turn-helix substructures. These differences are predicted to have little effect on the interactions of the aporepressor with tryptophan, holorepressor with operator DNA, or tandemly bound holorepressor dimers with one another. Although there is some variation observed at the dimer interface, interactions predicted to stabilize the interface are conserved. The phylogenetic relationships revealed by the TrpR amino acid sequence alignment agree with the results of others.
authors.affiliation: California Institute of Biological Research, La Jolla 92037.
authors.forename: D N
authors.initials: DN
authors.lastname: Arvidson
authors.forename: C G
authors.initials: CG
authors.lastname: Arvidson
authors.forename: C L
authors.initials: CL
authors.lastname: Lawson
authors.forename: J
authors.initials: J
authors.lastname: Miner
authors.forename: C
authors.initials: C
authors.lastname: Adams
authors.forename: P
authors.initials: P
authors.lastname: Youderian
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D001483:Base Sequence; D001665:Binding Sites; D017124:Conserved Sequence; D004269:DNA, Bacterial / Q000032:analysis; D004754:Enterobacter / Q000235:genetics; D004755:Enterobacteriaceae / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial / Q000235:genetics*; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009876:Operon / Q000235:genetics; D010802:Phylogeny; D011487:Protein Conformation; D012045:Regulatory Sequences, Nucleic Acid; D012097:Repressor Proteins / Q000235:genetics*; D012486:Salmonella typhimurium / Q000235:genetics; D016415:Sequence Alignment; D017384:Sequence Deletion / Q000235:genetics; D029968:Escherichia coli Proteins
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D012097:Repressor Proteins; C027190:TRPR protein, E coli; D029968:Escherichia coli Proteins