Conditionally expressed missense mutations: the basis for the unusual phenotype of an apparent trpD nonsense mutant of Salmonella typhimurium.
Tryptophan auxotroph trpo-28 is anomalous since preliminary mapping and suppression studies indicate the presence of a single amber nonsense mutation either late in trpE or early in trpD, but enzymological tests indicate the complete inactivation of both genes in this strain. Since the trpE and trpD genes are contiguous and encode the two subunits of a multifunctional enzyme complex, it was of interest to learn the mechanism of action of this apparent pleiotropic nonsense mutation. Our study has revealed that the phenotype of this strain derives not from a single mutation, but from the presenc and interaction of multiple mutations. Besides the recognized amber mutation (designated trpD28), this strain carries two additional, conditionally expressed missense mutations (designated trpE1651 and trpD1652). The trpD28 amber codon maps in the promoter-proximal region 1 of trpD and eliminates the glutamine amidotransferase activity of the bifunctional trpD polypeptide. The trpD1652 mutation maps in the promoter-distal region 2 of trpD and severely reduces (but does not eliminate) the phosphoribosyl transferase activity of the trpD polypeptide. The trpE1651 mutation maps in the anterior part of trpE and causes a rapid loss of activity of the trpE polypeptide, but only when it exists as an umcomplexed subunit. The existence of the two missense mutations escaped prior notice in standard recombinational tests since the nature of ech mutation is such that neither is detectable by the nutritional screens normally used in such tests unless an unsuppressed chain-terminating mutation, such as trpD28, is also present.
D002874:Chromosome Mapping; D002876:Chromosomes, Bacterial / Q000648:ultrastructure; D005815:Genetic Code; D005816:Genetic Complementation Test; D009154:Mutation; D009876:Operon; D010641:Phenotype; D011995:Recombination, Genetic; D012486:Salmonella typhimurium / Q000235:genetics*; D014364:Tryptophan / Q000235:genetics*
D014364:Tryptophan