Mutants of Salmonella typhimurium with an altered leucyl-transfer ribonucleic acid synthetase.
Two trifluoroleucine-resistant mutants of Salmonella typhimurium, strains CV69 and CV117, had an altered leucyl-transfer ribonucleic acid (tRNA) synthetase. The mutant enzymes had higher apparent K(m) values for leucine (ca. 10-fold) and lower specific activities (ca. twofold) than the parent enzyme when tested in crude extracts. Preparations of synthetase purified ca. 60-fold from the parent and strain CV117 differed sixfold in their leucine K(m) values. In addition, the mutant enzyme was inactivated faster than the parent enzyme at 50 C. The growth rates of strains CV69 and CV117 at 37 C were not significantly different from that of the parent, whereas at 42 C strain CV69 grew more slowly than the parent. Leucine-, valine-, and isoleucine-forming enzymes were partially derepressed when the mutants were grown in minimal medium; the addition of leucine repressed these enzymes to wild-type levels. During growth in minimal medium, the proportion of leucine tRNA that was charged in the mutants was about 75% of that in the parent. The properties of strain CV117 were shown to result from a single mutation located near gal at minute 18 on the genetic map. These studies suggest that leucyl-tRNA synthetase is involved in repression of the enzymes required for the synthesis of branched-chain amino acids.
D000255:Adenosine Triphosphate / Q000494:pharmacology; D001431:Bacteriological Techniques; D002247:Carbon Isotopes; D002874:Chromosome Mapping; D003227:Conjugation, Genetic; D003433:Crosses, Genetic; D003470:Culture Media; D004352:Drug Resistance, Microbial; D004794:Enzyme Repression; D005461:Fluorine / Q000494:pharmacology; D006836:Hydro-Lyases / Q000378:metabolism; D007532:Isoleucine / Q000096:biosynthesis; D007930:Leucine / Q000096:biosynthesis / Q000494:pharmacology; D008025:Ligases / Q000302:isolation & purification / Q000378:metabolism*; D008274:Magnesium / Q000494:pharmacology; D008956:Models, Chemical; D009154:Mutation; D012343:RNA, Transfer / Q000378:metabolism*; D012486:Salmonella typhimurium / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism; D014316:Tritium; D014633:Valine / Q000096:biosynthesis
D002247:Carbon Isotopes; D003470:Culture Media; D007532:Isoleucine; D014316:Tritium; D005461:Fluorine; D000255:Adenosine Triphosphate; D012343:RNA, Transfer; D006836:Hydro-Lyases; D008025:Ligases; D007930:Leucine; D014633:Valine; D008274:Magnesium