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. 1991 Apr;173(7):2341–2353. doi: 10.1128/jb.173.7.2341-2353.1991

Mutations replacing the leucine codons or altering the length of the amino acid-coding portion of the ilvGMEDA leader region of Escherichia coli.

J W Chen 1, E Harms 1, H E Umbarger 1
PMCID: PMC207787  PMID: 2007556

Abstract

The specificity of regulation by attenuation of the ilvGMEDA operon of Escherichia coli was examined by making alterations in the peptide-coding portion of the leader region. The effects of the alterations on attenuation control were monitored by operon fusions with the lacZ or cat gene. Substitution of the tandem leucine codons with arginine codons did not result in arginine control of attenuation even though the altered leader transcripts contained three consecutive arginine codons. Substitution of the single leucine codon with a proline codon at position 10 of the putative peptide, which had been shown to be important in the regulation of the Serratia marcescens ilv operon, did not result in control of attenuation by proline. Since the formation of neither proline nor arginine biosynthetic enzymes is regulated by attenuation control, the effect of tandem phenylalanine codons in place of the tandem leucine codons was examined and found not to result in control by phenylalanine supply. The latter failure may have been due to a configuration in the secondary structure of the protector stem of the leader transcript different from that of the wild-type transcript. The results of the study favored the idea that the lead ribosome does not initiate translation of the leader transcript until after the RNA polymerase has reached the pause site (117 bases into the leader region).

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Selected References

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

  1. Barnes W. M. DNA sequence from the histidine operon control region: seven histidine codons in a row. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4281–4285. doi: 10.1073/pnas.75.9.4281. [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. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  4. Chen J. W., Bennett D. C., Umbarger H. E. Specificity of attenuation control in the ilvGMEDA operon of Escherichia coli K-12. J Bacteriol. 1991 Apr;173(7):2328–2340. doi: 10.1128/jb.173.7.2328-2340.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Duggan D. E., Wechsler J. A. An assay for transaminase B enzyme activity in Escherichia coli K-12. Anal Biochem. 1973 Jan;51(1):67–79. doi: 10.1016/0003-2697(73)90453-3. [DOI] [PubMed] [Google Scholar]
  6. Edelmann P., Gallant J. Mistranslation in E. coli. Cell. 1977 Jan;10(1):131–137. doi: 10.1016/0092-8674(77)90147-7. [DOI] [PubMed] [Google Scholar]
  7. Gardner J. F. Regulation of the threonine operon: tandem threonine and isoleucine codons in the control region and translational control of transcription termination. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1706–1710. doi: 10.1073/pnas.76.4.1706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Garnier J., Osguthorpe D. J., Robson B. Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J Mol Biol. 1978 Mar 25;120(1):97–120. doi: 10.1016/0022-2836(78)90297-8. [DOI] [PubMed] [Google Scholar]
  9. Gayda D. J., Leathers T. D., Noti J. D., Smith F. J., Smith J. M., Subrahmanyam C. S., Umbarger H. E. Location of the multivalent control site for the ilvEDA operon of Escherichia coli. J Bacteriol. 1980 May;142(2):556–567. doi: 10.1128/jb.142.2.556-567.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gemmill R. M., Wessler S. R., Keller E. B., Calvo J. M. leu operon of Salmonella typhimurium is controlled by an attenuation mechanism. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4941–4945. doi: 10.1073/pnas.76.10.4941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Grosjean H., Fiers W. Preferential codon usage in prokaryotic genes: the optimal codon-anticodon interaction energy and the selective codon usage in efficiently expressed genes. Gene. 1982 Jun;18(3):199–209. doi: 10.1016/0378-1119(82)90157-3. [DOI] [PubMed] [Google Scholar]
  12. Grove C. L., Gunsalus R. P. Regulation of the aroH operon of Escherichia coli by the tryptophan repressor. J Bacteriol. 1987 May;169(5):2158–2164. doi: 10.1128/jb.169.5.2158-2164.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Harms E., Higgins E., Chen J. W., Umbarger H. E. Translational coupling between the ilvD and ilvA genes of Escherichia coli. J Bacteriol. 1988 Oct;170(10):4798–4807. doi: 10.1128/jb.170.10.4798-4807.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Hauser C. A., Sharp J. A., Hatfield L. K., Hatfield G. W. Pausing of RNA polymerase during in vitro transcription through the ilvB and ilvGEDA attenuator regions of Escherichia coli K12. J Biol Chem. 1985 Feb 10;260(3):1765–1770. [PubMed] [Google Scholar]
  16. Hsu J. H., Harms E., Umbarger H. E. Leucine regulation of the ilvGEDA operon of Serratia marcescens by attenuation is modulated by a single leucine codon. J Bacteriol. 1985 Oct;164(1):217–222. doi: 10.1128/jb.164.1.217-222.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Huang W. M., Ao S. Z., Casjens S., Orlandi R., Zeikus R., Weiss R., Winge D., Fang M. A persistent untranslated sequence within bacteriophage T4 DNA topoisomerase gene 60. Science. 1988 Feb 26;239(4843):1005–1012. doi: 10.1126/science.2830666. [DOI] [PubMed] [Google Scholar]
  18. Landick R., Yanofsky C., Choo K., Phung L. Replacement of the Escherichia coli trp operon attenuation control codons alters operon expression. J Mol Biol. 1990 Nov 5;216(1):25–37. doi: 10.1016/S0022-2836(05)80058-0. [DOI] [PubMed] [Google Scholar]
  19. Landick R., Yanofsky C. Stability of an RNA secondary structure affects in vitro transcription pausing in the trp operon leader region. J Biol Chem. 1984 Sep 25;259(18):11550–11555. [PubMed] [Google Scholar]
  20. Lawther R. P., Calhoun D. H., Gray J., Adams C. W., Hauser C. A., Hatfield G. W. DNA sequence fine-structure analysis of ilvG (IlvG+) mutations of Escherichia coli K-12. J Bacteriol. 1982 Jan;149(1):294–298. doi: 10.1128/jb.149.1.294-298.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lawther R. P., Hatfield G. W. Multivalent translational control of transcription termination at attenuator of ilvGEDA operon of Escherichia coli K-12. Proc Natl Acad Sci U S A. 1980 Apr;77(4):1862–1866. doi: 10.1073/pnas.77.4.1862. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lawther R. P., Lopes J. M., Ortuno M. J., White M. C. Analysis of regulation of the ilvGMEDA operon by using leader-attenuator-galK gene fusions. J Bacteriol. 1990 May;172(5):2320–2327. doi: 10.1128/jb.172.5.2320-2327.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lee F., Yanofsky C. Transcription termination at the trp operon attenuators of Escherichia coli and Salmonella typhimurium: RNA secondary structure and regulation of termination. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4365–4369. doi: 10.1073/pnas.74.10.4365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lopes J. M., Soliman N., Smith P. K., Lawther R. P. Transcriptional polarity enhances the contribution of the internal promoter, ilvEp, in the expression of the ilvGMEDA operon in wild-type Escherichia coli K12. Mol Microbiol. 1989 Aug;3(8):1039–1051. doi: 10.1111/j.1365-2958.1989.tb00254.x. [DOI] [PubMed] [Google Scholar]
  25. Nargang F. E., Subrahmanyam C. S., Umbarger H. E. Nucleotide sequence of ilvGEDA operon attenuator region of Escherichia coli. Proc Natl Acad Sci U S A. 1980 Apr;77(4):1823–1827. doi: 10.1073/pnas.77.4.1823. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Parker J. Errors and alternatives in reading the universal genetic code. Microbiol Rev. 1989 Sep;53(3):273–298. doi: 10.1128/mr.53.3.273-298.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rosner J. L. Formation, induction, and curing of bacteriophage P1 lysogens. Virology. 1972 Jun;48(3):679–689. doi: 10.1016/0042-6822(72)90152-3. [DOI] [PubMed] [Google Scholar]
  28. Sgaramella V., Ehrlich S. D., Bursztyn H., Lederberg J. Enhancement of transfecting activity of bacteriophage P22 DNA upon exonucleolytic erosion. J Mol Biol. 1976 Aug 25;105(4):587–602. doi: 10.1016/0022-2836(76)90237-0. [DOI] [PubMed] [Google Scholar]
  29. Simons R. W., Houman F., Kleckner N. Improved single and multicopy lac-based cloning vectors for protein and operon fusions. Gene. 1987;53(1):85–96. doi: 10.1016/0378-1119(87)90095-3. [DOI] [PubMed] [Google Scholar]
  30. Springer M., Trudel M., Graffe M., Plumbridge J., Fayat G., Mayaux J. F., Sacerdot C., Blanquet S., Grunberg-Manago M. Escherichia coli phenylalanyl-tRNA synthetase operon is controlled by attenuation in vivo. J Mol Biol. 1983 Dec 15;171(3):263–279. doi: 10.1016/0022-2836(83)90093-1. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. 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]
  33. 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]
  34. 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]
  35. Zoller M. J., Smith M. Oligonucleotide-directed mutagenesis of DNA fragments cloned into M13 vectors. Methods Enzymol. 1983;100:468–500. doi: 10.1016/0076-6879(83)00074-9. [DOI] [PubMed] [Google Scholar]
  36. Zuker M., Stiegler P. Optimal computer folding of large RNA sequences using thermodynamics and auxiliary information. Nucleic Acids Res. 1981 Jan 10;9(1):133–148. doi: 10.1093/nar/9.1.133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Zurawski G., Brown K., Killingly D., Yanofsky C. Nucleotide sequence of the leader region of the phenylalanine operon of Escherichia coli. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4271–4275. doi: 10.1073/pnas.75.9.4271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Zurawski G., Elseviers D., Stauffer G. V., Yanofsky C. Translational control of transcription termination at the attenuator of the Escherichia coli tryptophan operon. Proc Natl Acad Sci U S A. 1978 Dec;75(12):5988–5992. doi: 10.1073/pnas.75.12.5988. [DOI] [PMC free article] [PubMed] [Google Scholar]

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