tRNA(Trp) translation of leader peptide codon 12 and other factors that regulate expression of the tryptophanase operon.
Tryptophanase (tna) operon expression in Escherichia coli is induced by tryptophan. This response is mediated by features of a 319-base-pair leader region preceding the major structural genes of the operon. Translation of the coding region (tnaC) for a 24-amino-acid leader peptide is essential for induction. We have used site-directed mutagenesis to investigate the role of the single Trp codon, at position 12 in tnaC, in regulation of the operon. Codon 12 was changed to either a UAG or UGA stop codon or to a CGG arginine codon. Induction by tryptophan was eliminated by any of these changes. Studies with suppressor tRNAs indicated that tRNA(Trp) translation of codon 12 in tnaC is essential for induction of the operon. Reduction of tna expression by a miaA mutation supports a role for translation by tRNA(Trp) in regulation of the operon. Frameshift mutations and suppression that allows translation of tnaC to proceed beyond the normal stop codon result in constitutive tna operon expression. Deletion of a potential site for Rho factor utilization just beyond tnaC also results in partial constitutive expression. These studies suggest possible models for tryptophan induction of tna operon expression involving tRNA(Trp)-mediated frame shifting or readthrough at the tnaC stop codon.
Gollnick
P
P
Department of Biological Sciences, Stanford University, California 94305-5020.
Yanofsky
C
C
D002872:Chromosome Deletion; D003062:Codon; D015964:Gene Expression Regulation, Bacterial; D008190:Lyases / Q000235:genetics*; D009154:Mutation; D009690:Nucleic Acid Conformation; D009876:Operon; D014176:Protein Biosynthesis; D021382:Protein Sorting Signals / Q000235:genetics*; D012333:RNA, Messenger; D012345:RNA, Transfer, Amino Acid-Specific / Q000378:metabolism*; D012364:RNA, Transfer, Trp / Q000378:metabolism*; D013489:Suppression, Genetic; D014364:Tryptophan / Q000494:pharmacology; D014368:Tryptophanase / Q000235:genetics*
D003062:Codon; D021382:Protein Sorting Signals; D012333:RNA, Messenger; D012345:RNA, Transfer, Amino Acid-Specific; D012364:RNA, Transfer, Trp; D014364:Tryptophan; D008190:Lyases; D014368:Tryptophanase