title: Naturally occurring sites within the Shigella dysenteriae tryptophan operon severely limit tryptophan biosynthesis.
abstract: We investigated the structural, functional, and regulatory properties of the Shigella dysenteriae tryptophan (trp.) operon  in transduction hybrids in which the cysB-trp-region of Escherichia coli is replaced by the corresponding region from S. dysenteriae. Tryptophan biosynthesis was largely blocked in the hybrids, although the order of the structural genes was identical with that of E. coli. Nutritional tests and enzyme assays revealed that the hybrids produced a defective anthranilate synthetase (ASase). Deletion mapping identified two distinct sites in trpE, each of which was partially responsible for the instability and low activity of ASase. We also discovered a pleiotropic site trpP (S) that maps outside the structural gene region and is closely linked to the S. dysenteriae trp operator. trpP (S) reduced the rate of trp messenger ribonucleic acid synthesis, and consequently trp enzyme levels, 10-fold relative to wild-type E. coli. In recombinants in which the structural genes of E coli were under the control of the S. dysenteriae promoter, enzyme levels were also reduced 10-fold. In some fast-growing revertants of the original hybrids, the rates of trp messenger ribonucleic acid synthesis and levels of tryptophan synthetase were restored to values characteristic of wild-type E.coli. Thus, the Trp auxotrophy associated with the S dysenteriae trp operon derives from the combination of a defective ASase and decreased expression of the entire operon imposed by trpP (S).
mesh_terms: D000877:Anthranilate Phosphoribosyltransferase / Q000378:metabolism; D000878:Anthranilate Synthase / Q000378:metabolism; D002874:Chromosome Mapping; D004926:Escherichia coli / Q000201:enzymology / Q000378:metabolism; D005796:Genes; D006824:Hybridization, Genetic; D009154:Mutation; D009876:Operon; D012329:RNA, Bacterial / Q000096:biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis; D012762:Shigella dysenteriae / Q000201:enzymology / Q000378:metabolism*; D014161:Transduction, Genetic; D014364:Tryptophan / Q000096:biosynthesis*; D014367:Tryptophan Synthase / Q000378:metabolism
chemical_list: D012329:RNA, Bacterial; D012333:RNA, Messenger; D014364:Tryptophan; D000877:Anthranilate Phosphoribosyltransferase; D000878:Anthranilate Synthase; D014367:Tryptophan Synthase