Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1982 May;79(9):2812–2816. doi: 10.1073/pnas.79.9.2812

Agrobacterium Ti plasmid indoleacetic acid gene is required for crown gall oncogenesis

S-T Liu 1, K L Perry 1, C L Schardl 1, C I Kado 1,*
PMCID: PMC346296  PMID: 16593184

Abstract

A gene (iaaP) necessary for virulence and indoleacetic acid (IAA) production has been located on a nopaline Ti plasmid of Agrobacterium tumefaciens C58. iaaP function was established by using transformation to insert nopaline or octopine Ti plasmids into an avirulent, Ti plasmid-free mutant 1D1293-3 that was defective in IAA synthesis (iaaC-). The resulting transformants produced increased levels of IAA and virulence was restored. When these transformants were cured of their Ti plasmid, virulence and high IAA production levels were concomitantly lost. A Tn5 mutagenized TiC58 plasmid, deficient in the ability to direct increased synthesis of IAA, was inserted by transformation into mutant 1D1293-3. The resulting transformants 1D1293-3 (TiC58::Tn5) remained avirulent and iaaP-. Restriction analysis of the TiC58::Tn5 plasmid DNA identified the iaaP gene at 20.9 kilobases to the left of the T-DNA. A major aromatic-amino-acid aminotransferase is coded by the iaaC gene, but not by the iaaP gene. The possible reasons for the iaaP locus to be situated outside the T-DNA region are discussed.

Keywords: nopaline, octopine, oncogenesis, tryptophan aminotransferase

Full text

PDF
2812

Images in this article

Selected References

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

  1. Chilton M. D., Saiki R. K., Yadav N., Gordon M. P., Quetier F. T-DNA from Agrobacterium Ti plasmid is in the nuclear DNA fraction of crown gall tumor cells. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4060–4064. doi: 10.1073/pnas.77.7.4060. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Depicker A., De Wilde M., De Vos G., De Vos R., Van Montagu M., Schell J. Molecular cloning of overlapping segments of the nopaline Ti-plasmid pTiC58 as a means to restriction endonuclease mapping. Plasmid. 1980 Mar;3(2):193–211. doi: 10.1016/0147-619x(80)90109-2. [DOI] [PubMed] [Google Scholar]
  3. Garfinkel D. J., Nester E. W. Agrobacterium tumefaciens mutants affected in crown gall tumorigenesis and octopine catabolism. J Bacteriol. 1980 Nov;144(2):732–743. doi: 10.1128/jb.144.2.732-743.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Holsters M., Silva B., Van Vliet F., Genetello C., De Block M., Dhaese P., Depicker A., Inzé D., Engler G., Villarroel R. The functional organization of the nopaline A. tumefaciens plasmid pTiC58. Plasmid. 1980 Mar;3(2):212–230. doi: 10.1016/0147-619x(80)90110-9. [DOI] [PubMed] [Google Scholar]
  5. Holsters M., de Waele D., Depicker A., Messens E., van Montagu M., Schell J. Transfection and transformation of Agrobacterium tumefaciens. Mol Gen Genet. 1978 Jul 11;163(2):181–187. doi: 10.1007/BF00267408. [DOI] [PubMed] [Google Scholar]
  6. Jensen R. E., Zdybak W. T., Yasuda K., Chilton W. S. A useful synthesis of nopaline, a crown gall tumor metabolite. Biochem Biophys Res Commun. 1977 Apr 25;75(4):1066–1070. doi: 10.1016/0006-291x(77)91490-5. [DOI] [PubMed] [Google Scholar]
  7. KAPER J. M., VELDSTRA H. On the metabolism of tryptophan by Agrobacterium tumefaciens. Biochim Biophys Acta. 1958 Nov;30(2):401–420. doi: 10.1016/0006-3002(58)90065-9. [DOI] [PubMed] [Google Scholar]
  8. Kado C. I., Liu S. T. Rapid procedure for detection and isolation of large and small plasmids. J Bacteriol. 1981 Mar;145(3):1365–1373. doi: 10.1128/jb.145.3.1365-1373.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kline E. L., Brown C. S., Bankaitis V., Montefiori D. C., Craig K. Metabolite gene regulation of the L-arabinose operon in Escherichia coli with indoleacetic acid and other indole derivatives. Proc Natl Acad Sci U S A. 1980 Apr;77(4):1768–1772. doi: 10.1073/pnas.77.4.1768. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. LIN E. C., PITT B. M., CIVEN M., KNOX W. E. The assay of aromatic amino acid transaminations and keto acid oxidation by the enol borate-tautomerase method. J Biol Chem. 1958 Sep;233(3):668–673. [PubMed] [Google Scholar]
  11. LeBon J. M., Kado C. I., Rosenthal L. J., Chirikjian J. G. DNA modifying enzymes of Agrobacterium tumefaciens: effect of DNA topoisomerase, restriction endonuclease, and unique DNA endonuclease on plasmid and plant DNA. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4097–4101. doi: 10.1073/pnas.75.9.4097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lin B. C., Kado C. I. Studies on Agrobacterium tumefaciens. VIII. Avirulence induced by temperature and ethidium bromide. Can J Microbiol. 1977 Nov;23(11):1554–1561. doi: 10.1139/m77-229. [DOI] [PubMed] [Google Scholar]
  13. Liu S. T., Kado C. I. Indoleacetic acid production: a plasmid function of Agrobacterium tumefaciens C58. Biochem Biophys Res Commun. 1979 Sep 12;90(1):171–178. doi: 10.1016/0006-291x(79)91605-x. [DOI] [PubMed] [Google Scholar]
  14. Maniatis T., Jeffrey A., Kleid D. G. Nucleotide sequence of the rightward operator of phage lambda. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1184–1188. doi: 10.1073/pnas.72.3.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ooms G., Klapwijk P. M., Poulis J. A., Schilperoort R. A. Characterization of Tn904 insertions in octopine Ti plasmid mutants of Agrobacterium tumefaciens. J Bacteriol. 1980 Oct;144(1):82–91. doi: 10.1128/jb.144.1.82-91.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Paris C. G., Magasanik B. Tryptophan metabolism in Klebsiella aerogenes: regulation of the utilization of aromatic amino acids as sources of nitrogen. J Bacteriol. 1981 Jan;145(1):257–265. doi: 10.1128/jb.145.1.257-265.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Watson B., Currier T. C., Gordon M. P., Chilton M. D., Nester E. W. Plasmid required for virulence of Agrobacterium tumefaciens. J Bacteriol. 1975 Jul;123(1):255–264. doi: 10.1128/jb.123.1.255-264.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES