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Molecular characterization of the virulence gene virA of the Agrobacterium tumefaciens octopine Ti plasmid

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Abstract

The virulence loci play an essential role in tumor formation by Agrobacterium tumefaciens. Induction of vir gene expression by plant signal molecules is solely dependent on the virulence loci virA and virG. This study focused on the virA locus of the octopine type Ti plasmid pTi15955. The nucleic acid sequence of a 5.7-kilobase fragment encompassing virA was determined. Genetic analysis of this region revealed that virA contains one open reading frame coding for a protein of 91 639 daltons. Immunodetection with antibodies raised against a 35-kDa VirA fusion protein produced in E. coli identified by the VirA product in wild-type Agrobacterium cells. Moreover, it is shown that the VirA protein is located in the cytoplasmic membrane fraction of Agrobacterium. These data confirm the proposed regulatory function of VirA whereby VirA acts as a membrane sensor protein to identify plant signal molecules in the environment. The proposed sensory function of VirA strikingly resembles the function of the chemotaxis receptor proteins of E. coli.

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References

  1. Barker RF, Idler KB, Thompson DV, Kemp JD: Plant Mol Biol 2: 335–350 (1983).

    Google Scholar 

  2. Bevan MW, Chilton M-D: Ann Rev Genet 16: 357–384 (1982).

    Google Scholar 

  3. Birnboim HC, Doly J: Nucleic Acids Res 7: 1513–1523 (1979).

    Google Scholar 

  4. Burnette WM: Anal Biochem 112: 195–203 (1981).

    Google Scholar 

  5. Das A, Stachel S, Ebert P, Allenza P, Montoya A, Nester E: Nucleic Acids Res 14: 1355–1364 (1986).

    Google Scholar 

  6. DeFramond AJ, Barton KA, Chilton M-D: Biotechnology 1: 262–269 (1983).

    Google Scholar 

  7. DeMaagd RA, Lugtenberg B: J Bacteriol 167 1083–1085 (1986).

    Google Scholar 

  8. Devereux JR, Haeberli D, Smithies O: Nucleic Acids Res 12: 387–395 (1984).

    Google Scholar 

  9. DeVos G, DeBeuckeleer M, VanMontagu M, Schell J: Plasmid 6: 249–253 (1981).

    Google Scholar 

  10. Hall MN, Silhavy TJ: J Mol Biol 151: 1–15 (1981).

    Google Scholar 

  11. Halverson LJ, Stacey G: Microbiol Rev 50: 193–225 (1986).

    Google Scholar 

  12. Hille J, Klasen I, Schilperoort RA: Plasmid 7: 107–116 (1982).

    Google Scholar 

  13. Hille J, VanKan J, Klasen I, Schilperoort RA: J Bacteriol 154: 693–701 (1983).

    Google Scholar 

  14. Hille J, VanKan J, Schilperoort RA: J Bacteriol 158: 754–756 (1984).

    Google Scholar 

  15. Hoekema A, Hirsch PR, Hooykaas PJJ, Schilperoort RA: Nature 303: 179–180 (1983).

    Google Scholar 

  16. Hooykaas PJJ, Klapwijk PM, Nuti PM, Schilperoort RA, Rörsch A: J Gen Microbiol 98: 477–484 (1977).

    Google Scholar 

  17. Hooykaas PJJ, Hofker M, DemDulk-Ras H, Schilperoort RA: Plasmid 11: 195–205 (1984).

    Google Scholar 

  18. Hooykaas PJJ, Schilperoort RA: Adv Genet 22: 210–283 (1984).

    Google Scholar 

  19. Keleti G, Lederer WH: Handbook of Micromethods for the Biological Sciences. Van Nostrand Reinhold Co., New York (1974) pp 74–75.

    Google Scholar 

  20. Klee H, White FF, Iyer VN, Gordon MP, Nester EW: J Bacteriol 153: 878–883 (1983).

    Google Scholar 

  21. Knauf V, Yanofsky M, Monotya A, Nester E: J Bacteriol 160: 564–568 (1984).

    Google Scholar 

  22. Koekman BP, Hooykaas PJJ, Schilperoort RA: Plasmid 4: 184–195 (1980).

    Google Scholar 

  23. Krikos A, Mutoh N, Boyd A, Simon MI: Cell 33: 615–622 (1985).

    Google Scholar 

  24. Kyte J, Doolittle RF: J Mol Biol 157: 105–132 (1982).

    Google Scholar 

  25. Laemmli UK: Nature 227: 680–685 (1970).

    Google Scholar 

  26. Leemans J, Deblaere R, Willmitzer L, DeGreve H, Hernalsteens JP, VanMontagu M, Schell J: EMBO J 1: 147–152 (1982).

    Google Scholar 

  27. Leroux B, Yanofsky MF, Winans SC, Ward JE, Ziegler SF, Nester EW: EMBO J 6: 849–856 (1987).

    Google Scholar 

  28. Lipman DJ, Pearson WR: Science 227: 1435–1441 (1985).

    Google Scholar 

  29. Lundquist RC, Close TJ, Kado CI: Mol Gen Genet 193: 1–7 (1984).

    Google Scholar 

  30. Machida Y, Sakurai M, Kiyokawa S, Ubasawa A, Suzuki Y, Ikeda J-E: Proc Natl Acad Sci USA 81: 7495–7499 (1984).

    Google Scholar 

  31. Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: a Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).

    Google Scholar 

  32. Marsch JW, Erfle M, Wykes EJ: Gene 32: 481–485 (1984).

    Google Scholar 

  33. Maxam AM, Gilbert W: Methods Enzymol 65: 499–560 (1980).

    Google Scholar 

  34. McClure WR: Ann Rev Biochem 54: 171–204 (1985).

    Google Scholar 

  35. Melchers LS, Thompson DV, Idler KB, Schilperoort RA, Hooykaas PJJ: Nucleic Acids Res 14: 9933–9942 (1986).

    Google Scholar 

  36. Nester EW, Gordon MP, Amasino RM, Yanofsky MF: Ann Rev Plant Physiol 35: 387–413 (1984).

    Google Scholar 

  37. Ninfa AJ, Magasanik B: Proc Natl Acad Sci USA 83: 5909–5913 (1986).

    Google Scholar 

  38. Nixon BT, Ronson CW, Ausubel FM: Proc Natl Acad Sci USA 83: 7850–7854 (1986).

    Google Scholar 

  39. Okker RJH, Spaink H, Hille J, VanBrussel TAN, Lugtenberg B, Schilperoort RA. Nature 312: 564–566 (1984).

    Google Scholar 

  40. Osborn MJ, Gander JE, Parisi E, Carson J: J Biol Chem 247: 3962–3972 (1972).

    Google Scholar 

  41. Shine J, Dalgarno L: Proc Natl Acad Sci USA 77: 7117–7121 (1974).

    Google Scholar 

  42. Stachel SE, Messens E, VanMontagu M, Zambryski P: Nature 318: 624–629 (1985).

    Google Scholar 

  43. Stachel SE, Nester EW: EMBO J 5: 1445–1454 (1986).

    Google Scholar 

  44. Stachel SE, Zambryski PC: Cell 46: 325–333 (1986).

    Google Scholar 

  45. VonHeyne G: J Mol Biol 173: 243–251 (1984).

    Google Scholar 

  46. Winans SC, Ebert PR, Stachel SE, Gordon MP, Nester EW: Proc Natl Acad Sci USA 83: 8278–8282 (1986).

    Google Scholar 

  47. Yadav NS, Van derLeyden J, Bennett DR, Barnes WM, Chilton M-D: Proc Natl Acad Sci USA 79: 6322–6326 (1982).

    Google Scholar 

  48. Yamada T, Lee P-D, Kosuge T: Proc Natl Acad Sci USA 83: 8263–8267 (1986).

    Google Scholar 

  49. Yanofsky MF, Nester EW: J Bacteriol 168: 244–250 (1986).

    Google Scholar 

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Melchers, L.S., Thompson, D.V., Idler, K.B. et al. Molecular characterization of the virulence gene virA of the Agrobacterium tumefaciens octopine Ti plasmid. Plant Mol Biol 9, 635–645 (1987). https://doi.org/10.1007/BF00020539

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  • DOI: https://doi.org/10.1007/BF00020539

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