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. 1983 Jun;154(3):1054–1063. doi: 10.1128/jb.154.3.1054-1063.1983

IlvHI locus of Salmonella typhimurium.

C H Squires, M De Felice, C T Lago, J M Calvo
PMCID: PMC217575  PMID: 6189818

Abstract

In Escherichia coli K-12, the ilvHI locus codes for one of two acetohydroxy acid synthase isoenzymes. A region of the Salmonella typhimurium genome adjacent to the leucine operon was cloned on plasmid pBR322, yielding plasmids pCV47 and pCV49 (a shortened version of pCV47). This region contains DNA homologous to the E. coli ilvHI locus, as judged by hybridization experiments. Plasmid pCV47 did not confer isoleucine-valine prototrophy upon either E. coli or S. typhimurium strains lacking acetohydroxy acid synthase activity, suggesting that S. typhimurium lacks a functional ilvHI locus. However, isoleucine-valine prototrophs were readily isolated from such strains after mutagenesis with nitrosoguanidine. In one case we found that the Ilv+ phenotype resulted from an alteration in bacterial DNA on the plasmid (new plasmid designated pCV50). Furthermore, a new acetohydroxy acid synthase activity was observed in Ilv+ revertants; this enzyme was similar to E. coli acetohydroxy acid synthase III in its lack of activity at low pH. This new activity was correlated with the appearance in minicells of a new polypeptide having an approximate molecular weight of 61,000. Strains carrying either pCV49 or pCV50 produced a substantial amount of ilvHI-specific mRNA. These results, together with results from other laboratories, suggest that S. typhimurium has functional ilvB and ilvG genes and a cryptic ilvHI locus. E. coli K-12, on the other hand, has functional ilvB and ilvHI genes and a cryptic ilvG locus.

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Selected References

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  1. Alwine J. C., Kemp D. J., Stark G. R. Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5350–5354. doi: 10.1073/pnas.74.12.5350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bachmann B. J., Low K. B. Linkage map of Escherichia coli K-12, edition 6. Microbiol Rev. 1980 Mar;44(1):1–56. doi: 10.1128/mr.44.1.1-56.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bailey J. M., Davidson N. Methylmercury as a reversible denaturing agent for agarose gel electrophoresis. Anal Biochem. 1976 Jan;70(1):75–85. doi: 10.1016/s0003-2697(76)80049-8. [DOI] [PubMed] [Google Scholar]
  4. Berg C. M., Shaw K. J., Berg D. E. The ilvG gene is expressed in Escherichia coli K-12. Gene. 1980 Dec;12(1-2):165–170. doi: 10.1016/0378-1119(80)90028-1. [DOI] [PubMed] [Google Scholar]
  5. Calvo J. M., Freundlich M., Umbarger H. E. Regulation of branched-chain amino acid biosynthesis in Salmonella typhimurium: isolation of regulatory mutants. J Bacteriol. 1969 Mar;97(3):1272–1282. doi: 10.1128/jb.97.3.1272-1282.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. DEMEREC M., OHTA N. GENETIC ANALYSES OF SALMONELLA TYPHIMURIUM X ESCHERICHIA COLI HYBRIDS. Proc Natl Acad Sci U S A. 1964 Aug;52:317–323. doi: 10.1073/pnas.52.2.317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. De Felice M., Guardiola J., Esposito B., Iaccarino M. Structural genes for a newly recognized acetolactate synthase in Escherichia coli K-12. J Bacteriol. 1974 Dec;120(3):1068–1077. doi: 10.1128/jb.120.3.1068-1077.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. De Felice M., Guardiola J., Malorni M. C., Klopotowski T., Iaccarino M. Regulation of the pool size of valine in Escherichia coli K-12. J Bacteriol. 1974 Dec;120(3):1058–1067. doi: 10.1128/jb.120.3.1058-1067.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. De Felice M., Guardiola J., Schreil W., Levinthal M., Iaccarino M. Metabolic interlock between the acetolactate synthase isoenzymes and lysine biosynthesis in Escherichia coli K-12. Mol Gen Genet. 1977 Nov 4;156(1):9–16. doi: 10.1007/BF00272246. [DOI] [PubMed] [Google Scholar]
  10. De Felice M., Levinthal M., Iaccarino M., Guardiola J. Growth inhibition as a consequence of antagonism between related amino acids: effect of valine in Escherichia coli K-12. Microbiol Rev. 1979 Mar;43(1):42–58. doi: 10.1128/mr.43.1.42-58.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. De Felice M., Levinthal M. The acetohydroxy acid synthase III isoenzyme of Escherichia coli K-12: regulation of synthesis by leucine. Biochem Biophys Res Commun. 1977 Nov 7;79(1):82–87. doi: 10.1016/0006-291x(77)90063-8. [DOI] [PubMed] [Google Scholar]
  12. De Felice M., Squires C., Levinthal M., Guardiola J., Lamberti A., Iaccarino M. Growth inhibition of Escherichia coli K-12 by L-valine: a consequence of a regulatory pattern. Mol Gen Genet. 1977 Nov 4;156(1):1–7. doi: 10.1007/BF00272245. [DOI] [PubMed] [Google Scholar]
  13. Defez R., De Felice M. The metabolism of beta-glucosides in Escherichia coli K12. Ann Microbiol (Paris) 1982 May-Jun;133(3):347–350. [PubMed] [Google Scholar]
  14. Friden P., Newman T., Freundlich M. Nucleotide sequence of the ilvB promoter-regulatory region: a biosynthetic operon controlled by attenuation and cyclic AMP. Proc Natl Acad Sci U S A. 1982 Oct;79(20):6156–6160. doi: 10.1073/pnas.79.20.6156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Groves W. E., Davis F. C., Jr, Sells B. H. Spectrophotometric determination of microgram quantities of protein without nucleic acid interference. Anal Biochem. 1968 Feb;22(2):195–210. doi: 10.1016/0003-2697(68)90307-2. [DOI] [PubMed] [Google Scholar]
  16. Guardiola J., De Felice M., Iaccarino M. Mutant of Escherichia coli K-12 missing acetolactate synthase activity. J Bacteriol. 1974 Oct;120(1):536–538. doi: 10.1128/jb.120.1.536-538.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Guardiola J., De Felice M., Lamberti A., Iaccarino M. The acetolactate synthase isoenzymes of Escherichia coli K-12. Mol Gen Genet. 1977 Nov 4;156(1):17–25. doi: 10.1007/BF00272247. [DOI] [PubMed] [Google Scholar]
  18. Hertzberg K. M., Gemmill R., Jones J., Calvo J. M. Cloning of an EcoRI-generated fragment of the leucine operon of Salmonella typhimurium. Gene. 1980 Jan;8(2):135–152. doi: 10.1016/0378-1119(80)90033-5. [DOI] [PubMed] [Google Scholar]
  19. Lawther R. P., Calhoun D. H., Adams C. W., Hauser C. A., Gray J., Hatfield G. W. Molecular basis of valine resistance in Escherichia coli K-12. Proc Natl Acad Sci U S A. 1981 Feb;78(2):922–925. doi: 10.1073/pnas.78.2.922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Newman T. C., Levinthal M. A new map location for the ilvB locus of Escherichia coli. Genetics. 1980 Sep;96(1):59–77. doi: 10.1093/genetics/96.1.59. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Reeve J. Use of minicells for bacteriophage-directed polypeptide synthesis. Methods Enzymol. 1979;68:493–503. doi: 10.1016/0076-6879(79)68038-2. [DOI] [PubMed] [Google Scholar]
  22. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  23. Robinson C. L., Jackson J. H. New acetohydroxy acid synthase activity from mutational activation of a cryptic gene in Escherichia coli K-12. Mol Gen Genet. 1982;186(2):240–246. doi: 10.1007/BF00331856. [DOI] [PubMed] [Google Scholar]
  24. Shaw K. J., Berg C. M., Sobol T. J. Salmonella typhimurium mutants defective in acetohydroxy acid synthases I and II. J Bacteriol. 1980 Mar;141(3):1258–1263. doi: 10.1128/jb.141.3.1258-1263.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Smith D. R., Calvo J. M. Regulation of dihydrofolate reductase synthesis in Escherichia coli. Mol Gen Genet. 1979 Aug;175(1):31–38. doi: 10.1007/BF00267852. [DOI] [PubMed] [Google Scholar]
  26. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  27. Squires C. H., De Felice M., Wessler S. R., Calvo J. M. Physical characterization of the ilvHI operon of Escherichia coli K-12. J Bacteriol. 1981 Sep;147(3):797–804. doi: 10.1128/jb.147.3.797-804.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Stüber D., Bujard H. Organization of transcriptional signals in plasmids pBR322 and pACYC184. Proc Natl Acad Sci U S A. 1981 Jan;78(1):167–171. doi: 10.1073/pnas.78.1.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Subrahmanyam C. S., McCorkle G. M., Umbarger H. E. Physical location of the ilvO determinant in Escherichia coli K-12 deoxyribonucleic acid. J Bacteriol. 1980 May;142(2):547–555. doi: 10.1128/jb.142.2.547-555.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. de Felice M., Lago C. T., Squires C. H., Calvo J. M. Acetohydroxy acid synthase isoenzymes of Escherichia coli K12 and Salmonella typhimurium. Ann Microbiol (Paris) 1982 Mar-Apr;133(2):251–256. [PubMed] [Google Scholar]

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