Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1994 Jun;176(12):3765–3774. doi: 10.1128/jb.176.12.3765-3774.1994

Regulation of Caulobacter crescentus ilvBN gene expression.

J C Tarleton 1, J Malakooti 1, B Ely 1
PMCID: PMC205566  PMID: 8206855

Abstract

As part of an effort to determine the mechanisms employed by Caulobacter crescentus to regulate gene expression, the ilvBN genes encoding the two subunits of an acetohydroxy acid synthase (AHAS) have been characterized. Analysis of the DNA sequences indicated that the C. crescentus AHAS was highly homologous to AHAS isozymes from other organisms. S1 nuclease and primer extension studies demonstrated that transcription initiation occurred 172 bp upstream of the AHAS coding region. The region between the AHAS coding region and the transcription initiation site was shown to have the properties of a transcription attenuator. Deletion analysis of the region containing the stem-loop structure of the proposed attenuator resulted in the derepression of ilvBN expression. Thus, it appears that C. crescentus uses attenuation to regulate the expression of the ilvBN operon.

Full text

PDF
3765

Images in this article

Selected References

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

  1. Bagdasarian M., Lurz R., Rückert B., Franklin F. C., Bagdasarian M. M., Frey J., Timmis K. N. Specific-purpose plasmid cloning vectors. II. Broad host range, high copy number, RSF1010-derived vectors, and a host-vector system for gene cloning in Pseudomonas. Gene. 1981 Dec;16(1-3):237–247. doi: 10.1016/0378-1119(81)90080-9. [DOI] [PubMed] [Google Scholar]
  2. Bender R. A., Refson C. M., O'Neill E. A. Role of the flagellum in cell-cycle-dependent expression of bacteriophage receptor activity in Caulobacter crescentus. J Bacteriol. 1989 Feb;171(2):1035–1040. doi: 10.1128/jb.171.2.1035-1040.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chory J., Kaplan S. The in vitro transcription-translation of DNA and RNA templates by extracts of Rhodopseudomonas sphaeroides. Optimization and comparison of template specificity with Escherichia coli extracts and in vivo synthesis. J Biol Chem. 1982 Dec 25;257(24):15110–15121. [PubMed] [Google Scholar]
  4. Close T. J., Rodriguez R. L. Construction and characterization of the chloramphenicol-resistance gene cartridge: a new approach to the transcriptional mapping of extrachromosomal elements. Gene. 1982 Dec;20(2):305–316. doi: 10.1016/0378-1119(82)90048-8. [DOI] [PubMed] [Google Scholar]
  5. Coleman J., Green P. J., Inouye M. The use of RNAs complementary to specific mRNAs to regulate the expression of individual bacterial genes. Cell. 1984 Jun;37(2):429–436. doi: 10.1016/0092-8674(84)90373-8. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ely B. DNA sequence of the 3' end of the Caulobacter crescentus 16S rRNA gene. Nucleic Acids Res. 1992 Mar 25;20(6):1423–1423. doi: 10.1093/nar/20.6.1423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Falco S. C., Dumas K. S., Livak K. J. Nucleotide sequence of the yeast ILV2 gene which encodes acetolactate synthase. Nucleic Acids Res. 1985 Jun 11;13(11):4011–4027. doi: 10.1093/nar/13.11.4011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Friden P., Donegan J., Mullen J., Tsui P., Freundlich M., Eoyang L., Weber R., Silverman P. M. The ilvB locus of Escherichia coli K-12 is an operon encoding both subunits of acetohydroxyacid synthase I. Nucleic Acids Res. 1985 Jun 11;13(11):3979–3993. doi: 10.1093/nar/13.11.3979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. 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]
  13. Hauser C. A., Hatfield G. W. Attenuation of the ilvB operon by amino acids reflecting substrates or products of the ilvB gene product. Proc Natl Acad Sci U S A. 1984 Jan;81(1):76–79. doi: 10.1073/pnas.81.1.76. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hemsley A., Arnheim N., Toney M. D., Cortopassi G., Galas D. J. A simple method for site-directed mutagenesis using the polymerase chain reaction. Nucleic Acids Res. 1989 Aug 25;17(16):6545–6551. doi: 10.1093/nar/17.16.6545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Johnson R. C., Ely B. Isolation of spontaneously derived mutants of Caulobacter crescentus. Genetics. 1977 May;86(1):25–32. doi: 10.1093/genetics/86.1.25. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Landick R., Carey J., Yanofsky C. Translation activates the paused transcription complex and restores transcription of the trp operon leader region. Proc Natl Acad Sci U S A. 1985 Jul;82(14):4663–4667. doi: 10.1073/pnas.82.14.4663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. 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]
  20. Mazur B. J., Chui C. F., Smith J. K. Isolation and characterization of plant genes coding for acetolactate synthase, the target enzyme for two classes of herbicides. Plant Physiol. 1987 Dec;85(4):1110–1117. doi: 10.1104/pp.85.4.1110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pereira R. F., Ortuno M. J., Lawther R. P. Binding of integration host factor (IHF) to the ilvGp1 promoter of the ilvGMEDA operon of Escherichia coli K12. Nucleic Acids Res. 1988 Jul 11;16(13):5973–5989. doi: 10.1093/nar/16.13.5973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ross C. M., Winkler M. E. Structure of the Caulobacter crescentus trpFBA operon. J Bacteriol. 1988 Feb;170(2):757–768. doi: 10.1128/jb.170.2.757-768.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schloss J. V., Van Dyk D. E., Vasta J. F., Kutny R. M. Purification and properties of Salmonella typhimurium acetolactate synthase isozyme II from Escherichia coli HB101/pDU9. Biochemistry. 1985 Aug 27;24(18):4952–4959. doi: 10.1021/bi00339a034. [DOI] [PubMed] [Google Scholar]
  24. Schoenlein P. V., Gallman L. S., Ely B. Organization of the flaFG gene cluster and identification of two additional genes involved in flagellum biogenesis in Caulobacter crescentus. J Bacteriol. 1989 Mar;171(3):1544–1553. doi: 10.1128/jb.171.3.1544-1553.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Schoenlein P. V., Gallman L. S., Winkler M. E., Ely B. Nucleotide sequence of the Caulobacter crescentus flaF and flbT genes and an analysis of codon usage in organisms with G + C-rich genomes. Gene. 1990 Sep 1;93(1):17–25. doi: 10.1016/0378-1119(90)90130-j. [DOI] [PubMed] [Google Scholar]
  26. Shine J., Dalgarno L. Determinant of cistron specificity in bacterial ribosomes. Nature. 1975 Mar 6;254(5495):34–38. doi: 10.1038/254034a0. [DOI] [PubMed] [Google Scholar]
  27. Smith J. K., Schloss J. V., Mazur B. J. Functional expression of plant acetolactate synthase genes in Escherichia coli. Proc Natl Acad Sci U S A. 1989 Jun;86(11):4179–4183. doi: 10.1073/pnas.86.11.4179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Squires C. H., De Felice M., Devereux J., Calvo J. M. Molecular structure of ilvIH and its evolutionary relationship to ilvG in Escherichia coli K12. Nucleic Acids Res. 1983 Aug 11;11(15):5299–5313. doi: 10.1093/nar/11.15.5299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Tarleton J. C., Ely B. Isolation and characterization of ilvA, ilvBN, and ilvD mutants of Caulobacter crescentus. J Bacteriol. 1991 Feb;173(3):1259–1267. doi: 10.1128/jb.173.3.1259-1267.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wek R. C., Hauser C. A., Hatfield G. W. The nucleotide sequence of the ilvBN operon of Escherichia coli: sequence homologies of the acetohydroxy acid synthase isozymes. Nucleic Acids Res. 1985 Jun 11;13(11):3995–4010. doi: 10.1093/nar/13.11.3995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Winkler M. E., Schoenlein P. V., Ross C. M., Barrett J. T., Ely B. Genetic and physical analyses of Caulobacter crescentus trp genes. J Bacteriol. 1984 Oct;160(1):279–287. doi: 10.1128/jb.160.1.279-287.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yanofsky C. Attenuation in the control of expression of bacterial operons. Nature. 1981 Feb 26;289(5800):751–758. doi: 10.1038/289751a0. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES