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. 1990 Oct;172(10):5593–5601. doi: 10.1128/jb.172.10.5593-5601.1990

Indoleacetic acid operon of Pseudomonas syringae subsp. savastanoi: transcription analysis and promoter identification.

T D Gaffney 1, O da Costa e Silva 1, T Yamada 1, T Kosuge 1
PMCID: PMC526870  PMID: 2120185

Abstract

Expression of the indoleacetic acid (iaa) operon, which contributes to the virulence of the phytopathogenic bacterium Pseudomonas syringae subsp. savastanoi, was monitored by using broad-host-range lacZ reporter gene plasmids. A combination of translational (gene) fusions and transcriptional (operon) fusions of P. syringae subsp. savastanoi sequences to lacZ allowed localization of the iaa operon promoter. RNA recovered from P. syringae subsp. savastanoi strains was mapped with iaa operon-specific probes to precisely locate the transcription initiation site. When transcripts from an iaaM::lacZ fusion in Escherichia coli were analyzed, an identical transcription initiation site was observed. The DNA sequence of the iaa operon promoter closely resembled the consensus E. coli promoter sequence. We detected an active, constitutive level of indoleacetic acid biosynthetic gene expression during bacterial growth under a variety of conditions in the absence of host plant influence.

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Selected References

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

  1. Aiba H., Adhya S., de Crombrugghe B. Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem. 1981 Nov 25;256(22):11905–11910. [PubMed] [Google Scholar]
  2. BERTANI G. Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli. J Bacteriol. 1951 Sep;62(3):293–300. doi: 10.1128/jb.62.3.293-300.1951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clewell D. B., Helinski D. R. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159–1166. doi: 10.1073/pnas.62.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Comai L., Kosuge T. Cloning characterization of iaaM, a virulence determinant of Pseudomonas savastanoi. J Bacteriol. 1982 Jan;149(1):40–46. doi: 10.1128/jb.149.1.40-46.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Comai L., Kosuge T. Involvement of plasmid deoxyribonucleic acid in indoleacetic acid synthesis in Pseudomonas savastanoi. J Bacteriol. 1980 Aug;143(2):950–957. doi: 10.1128/jb.143.2.950-957.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Comai L., Kosuge T. Transposable element that causes mutations in a plant pathogenic Pseudomonas sp. J Bacteriol. 1983 Jun;154(3):1162–1167. doi: 10.1128/jb.154.3.1162-1167.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ditta G., Schmidhauser T., Yakobson E., Lu P., Liang X. W., Finlay D. R., Guiney D., Helinski D. R. Plasmids related to the broad host range vector, pRK290, useful for gene cloning and for monitoring gene expression. Plasmid. 1985 Mar;13(2):149–153. doi: 10.1016/0147-619x(85)90068-x. [DOI] [PubMed] [Google Scholar]
  9. Ditta G., Stanfield S., Corbin D., Helinski D. R. Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7347–7351. doi: 10.1073/pnas.77.12.7347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Débarbouillé M., Raibaud O. Expression of the Escherichia coli malPQ operon remains unaffected after drastic alteration of its promoter. J Bacteriol. 1983 Mar;153(3):1221–1227. doi: 10.1128/jb.153.3.1221-1227.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fett W. F., Osman S. F., Dunn M. F. Auxin production by plant-pathogenic pseudomonads and xanthomonads. Appl Environ Microbiol. 1987 Aug;53(8):1839–1845. doi: 10.1128/aem.53.8.1839-1845.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Glass N. L., Kosuge T. Cloning of the gene for indoleacetic acid-lysine synthetase from Pseudomonas syringae subsp. savastanoi. J Bacteriol. 1986 May;166(2):598–603. doi: 10.1128/jb.166.2.598-603.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hawley D. K., McClure W. R. Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res. 1983 Apr 25;11(8):2237–2255. doi: 10.1093/nar/11.8.2237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hutcheson S. W., Kosuge T. Regulation of 3-indoleacetic acid production in Pseudomonas syringae pv. savastanoi. Purification and properties of tryptophan 2-monooxygenase. J Biol Chem. 1985 May 25;260(10):6281–6287. [PubMed] [Google Scholar]
  15. Inouye S., Asai Y., Nakazawa A., Nakazawa T. Nucleotide sequence of a DNA segment promoting transcription in Pseudomonas putida. J Bacteriol. 1986 Jun;166(3):739–745. doi: 10.1128/jb.166.3.739-745.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Itoh Y., Soldati L., Stalon V., Falmagne P., Terawaki Y., Leisinger T., Haas D. Anabolic ornithine carbamoyltransferase of Pseudomonas aeruginosa: nucleotide sequence and transcriptional control of the argF structural gene. J Bacteriol. 1988 Jun;170(6):2725–2734. doi: 10.1128/jb.170.6.2725-2734.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. KING E. O., WARD M. K., RANEY D. E. Two simple media for the demonstration of pyocyanin and fluorescin. J Lab Clin Med. 1954 Aug;44(2):301–307. [PubMed] [Google Scholar]
  18. Kosuge T., Heskett M. G., Wilson E. E. Microbial synthesis and degradation of indole-3-acetic acid. I. The conversion of L-tryptophan to indole-3-acetamide by an enzyme system from Pseudomonas savastanoi. J Biol Chem. 1966 Aug 25;241(16):3738–3744. [PubMed] [Google Scholar]
  19. Palm C. J., Gaffney T., Kosuge T. Cotranscription of genes encoding indoleacetic acid production in Pseudomonas syringae subsp. savastanoi. J Bacteriol. 1989 Feb;171(2):1002–1009. doi: 10.1128/jb.171.2.1002-1009.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Powell G. K., Morris R. O. Nucleotide sequence and expression of a Pseudomonas savastanoi cytokinin biosynthetic gene: homology with Agrobacterium tumefaciens tmr and tzs loci. Nucleic Acids Res. 1986 Mar 25;14(6):2555–2565. doi: 10.1093/nar/14.6.2555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Shinshi H., Mohnen D., Meins F. Regulation of a plant pathogenesis-related enzyme: Inhibition of chitinase and chitinase mRNA accumulation in cultured tobacco tissues by auxin and cytokinin. Proc Natl Acad Sci U S A. 1987 Jan;84(1):89–93. doi: 10.1073/pnas.84.1.89. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Vieira J., Messing J. Production of single-stranded plasmid DNA. Methods Enzymol. 1987;153:3–11. doi: 10.1016/0076-6879(87)53044-0. [DOI] [PubMed] [Google Scholar]
  26. Yamada T., Lee P. D., Kosuge T. Insertion sequence elements of Pseudomonas savastanoi: Nucleotide sequence and homology with Agrobacterium tumefaciens transfer DNA. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8263–8267. doi: 10.1073/pnas.83.21.8263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Yamada T., Palm C. J., Brooks B., Kosuge T. Nucleotide sequences of the Pseudomonas savastanoi indoleacetic acid genes show homology with Agrobacterium tumefaciens T-DNA. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6522–6526. doi: 10.1073/pnas.82.19.6522. [DOI] [PMC free article] [PubMed] [Google Scholar]

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