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. 1988 Oct;170(10):4798–4807. doi: 10.1128/jb.170.10.4798-4807.1988

Translational coupling between the ilvD and ilvA genes of Escherichia coli.

E Harms 1, E Higgins 1, J W Chen 1, H E Umbarger 1
PMCID: PMC211523  PMID: 3049548

Abstract

The hypothesis that translation of the ilvD and ilvA genes of Escherichia coli may be linked has been examined in strains in which lacZ-ilvD protein fusions are translated in all three reading frames with respect to ilvD. In these strains, the nucleotide sequence was altered to obtain premature termination of ilvD translation, and in one strain translation termination of ilvD DNA occurred two bases downstream of the ilvA initiation codon. In the wild-type strain, the ilvD translation termination site was located two bases upstream of the ilvA start codon. In each of the mutant strains, expression of ilvA, as determined by the level of threonine deaminase activity, was strikingly lower than in the wild-type strain. The data suggest that expression of ilvD and ilvA is translationally coupled. By inserting a promoterless cat gene downstream of ilvA, it was shown that the differences in enzyme activity were not the result of differences in the amount of ilvA mRNA produced.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aksoy S., Squires C. L., Squires C. Translational coupling of the trpB and trpA genes in the Escherichia coli tryptophan operon. J Bacteriol. 1984 Feb;157(2):363–367. doi: 10.1128/jb.157.2.363-367.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bennett D. C., Umbarger H. E. Isolation and analysis of two Escherichia coli K-12 ilv attenuator deletion mutants with high-level constitutive expression of an ilv-lac fusion operon. J Bacteriol. 1984 Mar;157(3):839–845. doi: 10.1128/jb.157.3.839-845.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Berg C. M., Shaw K. J., Vender J., Borucka-Mankiewicz M. Physiological characterization of polar Tn5-induced isoleucine-valine auxotrophs in Escherichia coli K.12: evidence for an internal promoter in the ilvOGEDA operon. Genetics. 1979 Oct;93(2):308–319. [PMC free article] [PubMed] [Google Scholar]
  4. Blazey D. L., Burns R. O. Transcriptional activity of the transposable element Tn10 in the Salmonella typhimurium ilvGEDA operon. Proc Natl Acad Sci U S A. 1982 Aug;79(16):5011–5015. doi: 10.1073/pnas.79.16.5011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brosius J. Plasmid vectors for the selection of promoters. Gene. 1984 Feb;27(2):151–160. doi: 10.1016/0378-1119(84)90136-7. [DOI] [PubMed] [Google Scholar]
  6. Cone K. C., Steege D. A. Functional analysis of lac repressor restart sites in translational initiation and reinitiation. J Mol Biol. 1985 Dec 20;186(4):733–742. doi: 10.1016/0022-2836(85)90393-6. [DOI] [PubMed] [Google Scholar]
  7. Cone K. C., Steege D. A. Messenger RNA conformation and ribosome selection of translational reinitiation sites in the lac repressor mRNA. J Mol Biol. 1985 Dec 20;186(4):725–732. doi: 10.1016/0022-2836(85)90392-4. [DOI] [PubMed] [Google Scholar]
  8. Cox J. L., Cox B. J., Fidanza V., Calhoun D. H. The complete nucleotide sequence of the ilvGMEDA cluster of Escherichia coli K-12. Gene. 1987;56(2-3):185–198. doi: 10.1016/0378-1119(87)90136-3. [DOI] [PubMed] [Google Scholar]
  9. Das A., Yanofsky C. A ribosome binding site sequence is necessary for efficient expression of the distal gene of a translationally-coupled gene pair. Nucleic Acids Res. 1984 Jun 11;12(11):4757–4768. doi: 10.1093/nar/12.11.4757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Davis M. A., Simons R. W., Kleckner N. Tn10 protects itself at two levels from fortuitous activation by external promoters. Cell. 1985 Nov;43(1):379–387. doi: 10.1016/0092-8674(85)90043-1. [DOI] [PubMed] [Google Scholar]
  11. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hall M. N., Gabay J., Débarbouillé M., Schwartz M. A role for mRNA secondary structure in the control of translation initiation. Nature. 1982 Feb 18;295(5850):616–618. doi: 10.1038/295616a0. [DOI] [PubMed] [Google Scholar]
  13. Harms E., Hsu J. H., Subrahmanyam C. S., Umbarger H. E. Comparison of the regulatory regions of ilvGEDA operons from several enteric organisms. J Bacteriol. 1985 Oct;164(1):207–216. doi: 10.1128/jb.164.1.207-216.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Harms E., Umbarger H. E. Role of codon choice in the leader region of the ilvGMEDA operon of Serratia marcescens. J Bacteriol. 1987 Dec;169(12):5668–5677. doi: 10.1128/jb.169.12.5668-5677.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lawther R. P., Hatfield W. Biochemical characterization of an Escherichia coli hisT strain. J Bacteriol. 1977 Apr;130(1):552–557. doi: 10.1128/jb.130.1.552-557.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lawther R. P., Wek R. C., Lopes J. M., Pereira R., Taillon B. E., Hatfield G. W. The complete nucleotide sequence of the ilvGMEDA operon of Escherichia coli K-12. Nucleic Acids Res. 1987 Mar 11;15(5):2137–2155. doi: 10.1093/nar/15.5.2137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lopes J. M., Lawther R. P. Analysis and comparison of the internal promoter, pE, of the ilvGMEDA operons from Escherichia coli K-12 and Salmonella typhimurium. Nucleic Acids Res. 1986 Mar 25;14(6):2779–2798. doi: 10.1093/nar/14.6.2779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. McPheeters D. S., Christensen A., Young E. T., Stormo G., Gold L. Translational regulation of expression of the bacteriophage T4 lysozyme gene. Nucleic Acids Res. 1986 Jul 25;14(14):5813–5826. doi: 10.1093/nar/14.14.5813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Morgan W. D., Bear D. G., von Hippel P. H. Specificity of release by Escherichia coli transcription termination factor rho of nascent mRNA transcripts initiated at the lambda PR. J Biol Chem. 1984 Jul 10;259(13):8664–8671. [PubMed] [Google Scholar]
  20. Munson L. M., Stormo G. D., Niece R. L., Reznikoff W. S. lacZ translation initiation mutations. J Mol Biol. 1984 Aug 25;177(4):663–683. doi: 10.1016/0022-2836(84)90043-3. [DOI] [PubMed] [Google Scholar]
  21. Normark S., Bergström S., Edlund T., Grundström T., Jaurin B., Lindberg F. P., Olsson O. Overlapping genes. Annu Rev Genet. 1983;17:499–525. doi: 10.1146/annurev.ge.17.120183.002435. [DOI] [PubMed] [Google Scholar]
  22. Novick R. P., Clowes R. C., Cohen S. N., Curtiss R., 3rd, Datta N., Falkow S. Uniform nomenclature for bacterial plasmids: a proposal. Bacteriol Rev. 1976 Mar;40(1):168–189. doi: 10.1128/br.40.1.168-189.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Oppenheim D. S., Yanofsky C. Translational coupling during expression of the tryptophan operon of Escherichia coli. Genetics. 1980 Aug;95(4):785–795. doi: 10.1093/genetics/95.4.785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Prentki P., Krisch H. M. In vitro insertional mutagenesis with a selectable DNA fragment. Gene. 1984 Sep;29(3):303–313. doi: 10.1016/0378-1119(84)90059-3. [DOI] [PubMed] [Google Scholar]
  25. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schümperli D., McKenney K., Sobieski D. A., Rosenberg M. Translational coupling at an intercistronic boundary of the Escherichia coli galactose operon. Cell. 1982 Oct;30(3):865–871. doi: 10.1016/0092-8674(82)90291-4. [DOI] [PubMed] [Google Scholar]
  27. Shine J., Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. doi: 10.1073/pnas.71.4.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Smith J. M., Smith F. J., Umbarger H. E. Mutations affecting the formation of acetohydroxy acid synthase II in Escherichia coli K-12. Mol Gen Genet. 1979 Feb 1;169(3):299–314. doi: 10.1007/BF00382276. [DOI] [PubMed] [Google Scholar]
  29. Smith J. M., Smolin D. E., Umbarger H. E. Polarity and the regulation of the ilv gene cluster in Escherichia coli strain K-12. Mol Gen Genet. 1976 Oct 18;148(2):111–124. doi: 10.1007/BF00268374. [DOI] [PubMed] [Google Scholar]
  30. Sor F., Bolotin-Fukuhara M., Nomura M. Mutational alterations of translational coupling in the L11 ribosomal protein operon of Escherichia coli. J Bacteriol. 1987 Aug;169(8):3495–3507. doi: 10.1128/jb.169.8.3495-3507.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Steege D. A. 5'-Terminal nucleotide sequence of Escherichia coli lactose repressor mRNA: features of translational initiation and reinitiation sites. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4163–4167. doi: 10.1073/pnas.74.10.4163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. Subrahmanyam C. S., Noti J. D., Umbarger H. E. Regulation of ilvEDA expression occurs upstream of ilvG in Escherichia coli: additional evidence for an ilvGEDA operon. J Bacteriol. 1980 Oct;144(1):279–290. doi: 10.1128/jb.144.1.279-290.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Szentirmai A., Umbarger H. E. Isoleucine and valine metabolism of Escherichia coli. XIV. Effect of thiaisoleucine. J Bacteriol. 1968 May;95(5):1666–1671. doi: 10.1128/jb.95.5.1666-1671.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  36. Watson M. D., Wild J., Umbarger H. E. Positive control of ilvC expression in Escherichia coli K-12; identification and mapping of regulatory gene ilvY. J Bacteriol. 1979 Sep;139(3):1014–1020. doi: 10.1128/jb.139.3.1014-1020.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wechsler J. A., Adelberg E. A. Antipolarity in the ilv operon of Escherichia coli K-12. J Bacteriol. 1969 Jun;98(3):1179–1194. doi: 10.1128/jb.98.3.1179-1194.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wek R. C., Hatfield G. W. Examination of the internal promoter, PE, in the ilvGMEDA operon of E. coli K-12. Nucleic Acids Res. 1986 Mar 25;14(6):2763–2777. doi: 10.1093/nar/14.6.2763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  40. Yates J. L., Dean D., Strycharz W. A., Nomura M. E. coli ribosomal protein L10 inhibits translation of L10 and L7/L12 mRNAs by acting at a single site. Nature. 1981 Nov 12;294(5837):190–192. doi: 10.1038/294190a0. [DOI] [PubMed] [Google Scholar]

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