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. 1988 Dec;170(12):5479–5488. doi: 10.1128/jb.170.12.5479-5488.1988

Molecular analysis of a pathogenicity locus in Pseudomonas syringae pv. syringae.

P Mukhopadhyay 1, J Williams 1, D Mills 1
PMCID: PMC211641  PMID: 3192509

Abstract

One of the chromosomal regions of Pseudomonas syringae pv. syringae encoding pathogenicity factors had been mapped into a 3.9-kilobase-pair fragment in previous studies. Promoter probe analysis indicated the existence of a promoter near one end of the fragment. DNA sequencing of this fragment revealed the existence of a consensus promoter sequence in the region of the promoter activity and two open reading frames (ORFs) downstream. These ORFs, ORF1 and ORF2, encoded putative polypeptides of 40 and 83 kilodaltons, respectively. All ORF1::Tn5 as well as ORF2::Tn5 mutant strains were nonpathogenic on susceptible host bean plants and were unable to elicit hypersensitive reactions on nonhost tobacco plants. The deduced amino acid sequence of the 83-kilodalton polypeptide contained features characteristic of known integral membrane proteins. Fusion of the lacZ gene to ORF2 led to the expression of a hybrid protein inducible in Escherichia coli. The functions of the putative proteins encoded by ORF1 and ORF2 are unknown at present.

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Selected References

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  1. Alff-Steinberger C. Evidence for a coding pattern on the non-coding strand of the E. coli genome. Nucleic Acids Res. 1984 Mar 12;12(5):2235–2241. doi: 10.1093/nar/12.5.2235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berg D. E., Weiss A., Crossland L. Polarity of Tn5 insertion mutations in Escherichia coli. J Bacteriol. 1980 May;142(2):439–446. doi: 10.1128/jb.142.2.439-446.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boucher C. A., Van Gijsegem F., Barberis P. A., Arlat M., Zischek C. Pseudomonas solanacearum genes controlling both pathogenicity on tomato and hypersensitivity on tobacco are clustered. J Bacteriol. 1987 Dec;169(12):5626–5632. doi: 10.1128/jb.169.12.5626-5632.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Coles L. S., Wells J. R. An H1 histone gene-specific 5' element and evolution of H1 and H5 genes. Nucleic Acids Res. 1985 Jan 25;13(2):585–594. doi: 10.1093/nar/13.2.585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Conrad B., Mount D. W. Microcomputer programs for DNA sequence analysis. Nucleic Acids Res. 1982 Jan 11;10(1):31–38. doi: 10.1093/nar/10.1.31. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cuppels D. A. Generation and Characterization of Tn5 Insertion Mutations in Pseudomonas syringae pv. tomato. Appl Environ Microbiol. 1986 Feb;51(2):323–327. doi: 10.1128/aem.51.2.323-327.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Deininger C. A., Mueller G. M., Wolber P. K. Immunological characterization of ice nucleation proteins from Pseudomonas syringae, Pseudomonas fluorescens, and Erwinia herbicola. J Bacteriol. 1988 Feb;170(2):669–675. doi: 10.1128/jb.170.2.669-675.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Engelman D. M., Steitz T. A., Goldman A. Identifying nonpolar transbilayer helices in amino acid sequences of membrane proteins. Annu Rev Biophys Biophys Chem. 1986;15:321–353. doi: 10.1146/annurev.bb.15.060186.001541. [DOI] [PubMed] [Google Scholar]
  9. Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. doi: 10.1016/0378-1119(84)90153-7. [DOI] [PubMed] [Google Scholar]
  10. Hopp T. P., Woods K. R. Prediction of protein antigenic determinants from amino acid sequences. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3824–3828. doi: 10.1073/pnas.78.6.3824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  12. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  13. Langley K. E., Villarejo M. R., Fowler A. V., Zamenhof P. J., Zabin I. Molecular basis of beta-galactosidase alpha-complementation. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1254–1257. doi: 10.1073/pnas.72.4.1254. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lindgren P. B., Peet R. C., Panopoulos N. J. Gene cluster of Pseudomonas syringae pv. "phaseolicola" controls pathogenicity of bean plants and hypersensitivity of nonhost plants. J Bacteriol. 1986 Nov;168(2):512–522. doi: 10.1128/jb.168.2.512-522.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. McKenney K., Shimatake H., Court D., Schmeissner U., Brady C., Rosenberg M. A system to study promoter and terminator signals recognized by Escherichia coli RNA polymerase. Gene Amplif Anal. 1981;2:383–415. [PubMed] [Google Scholar]
  16. Mizusawa S., Nishimura S., Seela F. Improvement of the dideoxy chain termination method of DNA sequencing by use of deoxy-7-deazaguanosine triphosphate in place of dGTP. Nucleic Acids Res. 1986 Feb 11;14(3):1319–1324. doi: 10.1093/nar/14.3.1319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mount D. W., Conrad B. Microcomputer programs for graphic analysis of nucleic acid and protein sequences. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):811–817. doi: 10.1093/nar/12.1part2.811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Murooka Y., Iwamoto H., Hamamoto A., Yamauchi T. Efficient transformation of phytopathogenic strains of Xanthomonas species. J Bacteriol. 1987 Sep;169(9):4406–4409. doi: 10.1128/jb.169.9.4406-4409.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nathans J., Hogness D. S. Isolation, sequence analysis, and intron-exon arrangement of the gene encoding bovine rhodopsin. Cell. 1983 Oct;34(3):807–814. doi: 10.1016/0092-8674(83)90537-8. [DOI] [PubMed] [Google Scholar]
  20. Niepold F., Anderson D., Mills D. Cloning determinants of pathogenesis from Pseudomonas syringae pathovar syringae. Proc Natl Acad Sci U S A. 1985 Jan;82(2):406–410. doi: 10.1073/pnas.82.2.406. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Osbourn A. E., Barber C. E., Daniels M. J. Identification of plant-induced genes of the bacterial pathogen Xanthomonas campestris pathovar campestris using a promoter-probe plasmid. EMBO J. 1987 Jan;6(1):23–28. doi: 10.1002/j.1460-2075.1987.tb04713.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Remaut E., Tsao H., Fiers W. Improved plasmid vectors with a thermoinducible expression and temperature-regulated runaway replication. Gene. 1983 Apr;22(1):103–113. doi: 10.1016/0378-1119(83)90069-0. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. 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]
  26. Sutcliffe J. G. Nucleotide sequence of the ampicillin resistance gene of Escherichia coli plasmid pBR322. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3737–3741. doi: 10.1073/pnas.75.8.3737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Xu P. L., Leong S., Sequeira L. Molecular cloning of genes that specify virulence in Pseudomonas solanacearum. J Bacteriol. 1988 Feb;170(2):617–622. doi: 10.1128/jb.170.2.617-622.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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]
  29. 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]

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