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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Barker RF, Idler KB, Thompson DV, Kemp JD: Plant Mol Biol 2: 335–350 (1983).
Bevan MW, Chilton M-D: Ann Rev Genet 16: 357–384 (1982).
Birnboim HC, Doly J: Nucleic Acids Res 7: 1513–1523 (1979).
Burnette WM: Anal Biochem 112: 195–203 (1981).
Das A, Stachel S, Ebert P, Allenza P, Montoya A, Nester E: Nucleic Acids Res 14: 1355–1364 (1986).
DeFramond AJ, Barton KA, Chilton M-D: Biotechnology 1: 262–269 (1983).
DeMaagd RA, Lugtenberg B: J Bacteriol 167 1083–1085 (1986).
Devereux JR, Haeberli D, Smithies O: Nucleic Acids Res 12: 387–395 (1984).
DeVos G, DeBeuckeleer M, VanMontagu M, Schell J: Plasmid 6: 249–253 (1981).
Hall MN, Silhavy TJ: J Mol Biol 151: 1–15 (1981).
Halverson LJ, Stacey G: Microbiol Rev 50: 193–225 (1986).
Hille J, Klasen I, Schilperoort RA: Plasmid 7: 107–116 (1982).
Hille J, VanKan J, Klasen I, Schilperoort RA: J Bacteriol 154: 693–701 (1983).
Hille J, VanKan J, Schilperoort RA: J Bacteriol 158: 754–756 (1984).
Hoekema A, Hirsch PR, Hooykaas PJJ, Schilperoort RA: Nature 303: 179–180 (1983).
Hooykaas PJJ, Klapwijk PM, Nuti PM, Schilperoort RA, Rörsch A: J Gen Microbiol 98: 477–484 (1977).
Hooykaas PJJ, Hofker M, DemDulk-Ras H, Schilperoort RA: Plasmid 11: 195–205 (1984).
Hooykaas PJJ, Schilperoort RA: Adv Genet 22: 210–283 (1984).
Keleti G, Lederer WH: Handbook of Micromethods for the Biological Sciences. Van Nostrand Reinhold Co., New York (1974) pp 74–75.
Klee H, White FF, Iyer VN, Gordon MP, Nester EW: J Bacteriol 153: 878–883 (1983).
Knauf V, Yanofsky M, Monotya A, Nester E: J Bacteriol 160: 564–568 (1984).
Koekman BP, Hooykaas PJJ, Schilperoort RA: Plasmid 4: 184–195 (1980).
Krikos A, Mutoh N, Boyd A, Simon MI: Cell 33: 615–622 (1985).
Kyte J, Doolittle RF: J Mol Biol 157: 105–132 (1982).
Laemmli UK: Nature 227: 680–685 (1970).
Leemans J, Deblaere R, Willmitzer L, DeGreve H, Hernalsteens JP, VanMontagu M, Schell J: EMBO J 1: 147–152 (1982).
Leroux B, Yanofsky MF, Winans SC, Ward JE, Ziegler SF, Nester EW: EMBO J 6: 849–856 (1987).
Lipman DJ, Pearson WR: Science 227: 1435–1441 (1985).
Lundquist RC, Close TJ, Kado CI: Mol Gen Genet 193: 1–7 (1984).
Machida Y, Sakurai M, Kiyokawa S, Ubasawa A, Suzuki Y, Ikeda J-E: Proc Natl Acad Sci USA 81: 7495–7499 (1984).
Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: a Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).
Marsch JW, Erfle M, Wykes EJ: Gene 32: 481–485 (1984).
Maxam AM, Gilbert W: Methods Enzymol 65: 499–560 (1980).
McClure WR: Ann Rev Biochem 54: 171–204 (1985).
Melchers LS, Thompson DV, Idler KB, Schilperoort RA, Hooykaas PJJ: Nucleic Acids Res 14: 9933–9942 (1986).
Nester EW, Gordon MP, Amasino RM, Yanofsky MF: Ann Rev Plant Physiol 35: 387–413 (1984).
Ninfa AJ, Magasanik B: Proc Natl Acad Sci USA 83: 5909–5913 (1986).
Nixon BT, Ronson CW, Ausubel FM: Proc Natl Acad Sci USA 83: 7850–7854 (1986).
Okker RJH, Spaink H, Hille J, VanBrussel TAN, Lugtenberg B, Schilperoort RA. Nature 312: 564–566 (1984).
Osborn MJ, Gander JE, Parisi E, Carson J: J Biol Chem 247: 3962–3972 (1972).
Shine J, Dalgarno L: Proc Natl Acad Sci USA 77: 7117–7121 (1974).
Stachel SE, Messens E, VanMontagu M, Zambryski P: Nature 318: 624–629 (1985).
Stachel SE, Nester EW: EMBO J 5: 1445–1454 (1986).
Stachel SE, Zambryski PC: Cell 46: 325–333 (1986).
VonHeyne G: J Mol Biol 173: 243–251 (1984).
Winans SC, Ebert PR, Stachel SE, Gordon MP, Nester EW: Proc Natl Acad Sci USA 83: 8278–8282 (1986).
Yadav NS, Van derLeyden J, Bennett DR, Barnes WM, Chilton M-D: Proc Natl Acad Sci USA 79: 6322–6326 (1982).
Yamada T, Lee P-D, Kosuge T: Proc Natl Acad Sci USA 83: 8263–8267 (1986).
Yanofsky MF, Nester EW: J Bacteriol 168: 244–250 (1986).
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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