Abstract
Early studies on Agrobacterium tumefaciens showed that development of tumors on plants following infection by A. tumefaciens was optimal at temperatures around 22 degrees C and did not occur at temperatures above 29 degrees C. To assess whether this inability to induce tumors is due to a defect in the T-DNA transfer machinery, mobilization of an incompatibility group Q (IncQ) plasmid by the T-DNA transfer machinery of A. tumefaciens was tested at various temperatures. Optimal transfer occurred when matings were performed at 19 degrees C, and transfer was not seen when matings were incubated above 28 degrees C. Transfer of the IncQ plasmid was dependent upon induction of the virB and virD operons by acetosyringone but was not dependent upon induction of the tra genes by octopine. However, alterations in the level of vir gene induction could not account for the decrease in transfer with increasing temperature. A. tumefaciens did successfully mobilize IncQ plasmids at higher temperatures when alternative transfer machineries were provided. Thus, the defect in transfer at high temperature is apparently in the T-DNA transfer machinery itself. As these data correlate with earlier tumorigenesis studies, we propose that tumor suppression at higher temperatures results from a T-DNA transfer machinery which does not function properly.
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Selected References
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- Ankenbauer R. G., Best E. A., Palanca C. A., Nester E. W. Mutants of the Agrobacterium tumefaciens virA gene exhibiting acetosyringone-independent expression of the vir regulon. Mol Plant Microbe Interact. 1991 Jul-Aug;4(4):400–406. doi: 10.1094/mpmi-4-400. [DOI] [PubMed] [Google Scholar]
- BRAUN A. C. Conditioning of the host cell as a factor in the transformation process in crown gall. Growth. 1952 Jun;16(2):65–74. [PubMed] [Google Scholar]
- Beijersbergen A., Dulk-Ras A. D., Schilperoort R. A., Hooykaas P. J. Conjugative Transfer by the Virulence System of Agrobacterium tumefaciens. Science. 1992 May 29;256(5061):1324–1327. doi: 10.1126/science.256.5061.1324. [DOI] [PubMed] [Google Scholar]
- Beijersbergen A., Smith S. J., Hooykaas P. J. Localization and topology of VirB proteins of Agrobacterium tumefaciens. Plasmid. 1994 Sep;32(2):212–218. doi: 10.1006/plas.1994.1057. [DOI] [PubMed] [Google Scholar]
- Berger B. R., Christie P. J. Genetic complementation analysis of the Agrobacterium tumefaciens virB operon: virB2 through virB11 are essential virulence genes. J Bacteriol. 1994 Jun;176(12):3646–3660. doi: 10.1128/jb.176.12.3646-3660.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Berger B. R., Christie P. J. The Agrobacterium tumefaciens virB4 gene product is an essential virulence protein requiring an intact nucleoside triphosphate-binding domain. J Bacteriol. 1993 Mar;175(6):1723–1734. doi: 10.1128/jb.175.6.1723-1734.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bilge S. S., Apostol J. M., Jr, Fullner K. J., Moseley S. L. Transcriptional organization of the F1845 fimbrial adhesin determinant of Escherichia coli. Mol Microbiol. 1993 Mar;7(6):993–1006. doi: 10.1111/j.1365-2958.1993.tb01191.x. [DOI] [PubMed] [Google Scholar]
- Bradley D. E., Whelan J. Conjugation systems of IncT plasmids. J Gen Microbiol. 1985 Oct;131(10):2665–2671. doi: 10.1099/00221287-131-10-2665. [DOI] [PubMed] [Google Scholar]
- Braun A. C. A Physiological Basis for Autonomous Growth of the Crown-Gall Tumor Cell. Proc Natl Acad Sci U S A. 1958 Apr;44(4):344–349. doi: 10.1073/pnas.44.4.344. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bundock P., den Dulk-Ras A., Beijersbergen A., Hooykaas P. J. Trans-kingdom T-DNA transfer from Agrobacterium tumefaciens to Saccharomyces cerevisiae. EMBO J. 1995 Jul 3;14(13):3206–3214. doi: 10.1002/j.1460-2075.1995.tb07323.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cangelosi G. A., Best E. A., Martinetti G., Nester E. W. Genetic analysis of Agrobacterium. Methods Enzymol. 1991;204:384–397. doi: 10.1016/0076-6879(91)04020-o. [DOI] [PubMed] [Google Scholar]
- Christie P. J., Ward J. E., Jr, Gordon M. P., Nester E. W. A gene required for transfer of T-DNA to plants encodes an ATPase with autophosphorylating activity. Proc Natl Acad Sci U S A. 1989 Dec;86(24):9677–9681. doi: 10.1073/pnas.86.24.9677. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cook D. M., Farrand S. K. The oriT region of the Agrobacterium tumefaciens Ti plasmid pTiC58 shares DNA sequence identity with the transfer origins of RSF1010 and RK2/RP4 and with T-region borders. J Bacteriol. 1992 Oct;174(19):6238–6246. doi: 10.1128/jb.174.19.6238-6246.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Doty S. L., Chang M., Nester E. W. The chromosomal virulence gene, chvE, of Agrobacterium tumefaciens is regulated by a LysR family member. J Bacteriol. 1993 Dec;175(24):7880–7886. doi: 10.1128/jb.175.24.7880-7886.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Farrand S. K., O'Morchoe S. P., McCutchan J. Construction of an Agrobacterium tumefaciens C58 recA mutant. J Bacteriol. 1989 Oct;171(10):5314–5321. doi: 10.1128/jb.171.10.5314-5321.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fullner K. J., Stephens K. M., Nester E. W. An essential virulence protein of Agrobacterium tumefaciens, VirB4, requires an intact mononucleotide binding domain to function in transfer of T-DNA. Mol Gen Genet. 1994 Dec 15;245(6):704–715. doi: 10.1007/BF00297277. [DOI] [PubMed] [Google Scholar]
- 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]
- Jin S., Song Y. N., Deng W. Y., Gordon M. P., Nester E. W. The regulatory VirA protein of Agrobacterium tumefaciens does not function at elevated temperatures. J Bacteriol. 1993 Nov;175(21):6830–6835. doi: 10.1128/jb.175.21.6830-6835.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kado C. I. Promiscuous DNA transfer system of Agrobacterium tumefaciens: role of the virB operon in sex pilus assembly and synthesis. Mol Microbiol. 1994 Apr;12(1):17–22. doi: 10.1111/j.1365-2958.1994.tb00990.x. [DOI] [PubMed] [Google Scholar]
- Koukolíková-Nicola Z., Raineri D., Stephens K., Ramos C., Tinland B., Nester E. W., Hohn B. Genetic analysis of the virD operon of Agrobacterium tumefaciens: a search for functions involved in transport of T-DNA into the plant cell nucleus and in T-DNA integration. J Bacteriol. 1993 Feb;175(3):723–731. doi: 10.1128/jb.175.3.723-731.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Labes M., Pühler A., Simon R. A new family of RSF1010-derived expression and lac-fusion broad-host-range vectors for gram-negative bacteria. Gene. 1990 Apr 30;89(1):37–46. doi: 10.1016/0378-1119(90)90203-4. [DOI] [PubMed] [Google Scholar]
- Lessl M., Lanka E. Common mechanisms in bacterial conjugation and Ti-mediated T-DNA transfer to plant cells. Cell. 1994 May 6;77(3):321–324. doi: 10.1016/0092-8674(94)90146-5. [DOI] [PubMed] [Google Scholar]
- Maher D., Sherburne R., Taylor D. E. H-pilus assembly kinetics determined by electron microscopy. J Bacteriol. 1993 Apr;175(8):2175–2183. doi: 10.1128/jb.175.8.2175-2183.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Okamoto S., Toyoda-Yamamoto A., Ito K., Takebe I., Machida Y. Localization and orientation of the VirD4 protein of Agrobacterium tumefaciens in the cell membrane. Mol Gen Genet. 1991 Aug;228(1-2):24–32. doi: 10.1007/BF00282443. [DOI] [PubMed] [Google Scholar]
- Piper K. R., Beck von Bodman S., Farrand S. K. Conjugation factor of Agrobacterium tumefaciens regulates Ti plasmid transfer by autoinduction. Nature. 1993 Apr 1;362(6419):448–450. doi: 10.1038/362448a0. [DOI] [PubMed] [Google Scholar]
- Pohlman R. F., Genetti H. D., Winans S. C. Common ancestry between IncN conjugal transfer genes and macromolecular export systems of plant and animal pathogens. Mol Microbiol. 1994 Nov;14(4):655–668. doi: 10.1111/j.1365-2958.1994.tb01304.x. [DOI] [PubMed] [Google Scholar]
- Prentki P., Krisch H. M. In vitro insertional mutagenesis with a selectable DNA fragment. Gene. 1984 Sep;29(3):303–313. doi: 10.1016/0378-1119(84)90059-3. [DOI] [PubMed] [Google Scholar]
- Sciaky D., Montoya A. L., Chilton M. D. Fingerprints of Agrobacterium Ti plasmids. Plasmid. 1978 Feb;1(2):238–253. doi: 10.1016/0147-619x(78)90042-2. [DOI] [PubMed] [Google Scholar]
- Shirasu K., Kado C. I. Membrane location of the Ti plasmid VirB proteins involved in the biosynthesis of a pilin-like conjugative structure on Agrobacterium tumefaciens. FEMS Microbiol Lett. 1993 Aug 1;111(2-3):287–294. doi: 10.1111/j.1574-6968.1993.tb06400.x. [DOI] [PubMed] [Google Scholar]
- Shirasu K., Koukolíková-Nicola Z., Hohn B., Kado C. I. An inner-membrane-associated virulence protein essential for T-DNA transfer from Agrobacterium tumefaciens to plants exhibits ATPase activity and similarities to conjugative transfer genes. Mol Microbiol. 1994 Feb;11(3):581–588. doi: 10.1111/j.1365-2958.1994.tb00338.x. [DOI] [PubMed] [Google Scholar]
- Stachel S. E., Nester E. W. The genetic and transcriptional organization of the vir region of the A6 Ti plasmid of Agrobacterium tumefaciens. EMBO J. 1986 Jul;5(7):1445–1454. doi: 10.1002/j.1460-2075.1986.tb04381.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Taylor D. E., Levine J. G. Studies of temperature-sensitive transfer and maintenance of H incompatibility group plasmids. J Gen Microbiol. 1980 Feb;116(2):475–484. doi: 10.1099/00221287-116-2-475. [DOI] [PubMed] [Google Scholar]
- Tempé J., Petit A., Holsters M., Montagu M., Schell J. Thermosensitive step associated with transfer of the Ti plasmid during conjugation: Possible relation to transformation in crown gall. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2848–2849. doi: 10.1073/pnas.74.7.2848. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thorstenson Y. R., Kuldau G. A., Zambryski P. C. Subcellular localization of seven VirB proteins of Agrobacterium tumefaciens: implications for the formation of a T-DNA transport structure. J Bacteriol. 1993 Aug;175(16):5233–5241. doi: 10.1128/jb.175.16.5233-5241.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thorstenson Y. R., Zambryski P. C. The essential virulence protein VirB8 localizes to the inner membrane of Agrobacterium tumefaciens. J Bacteriol. 1994 Mar;176(6):1711–1717. doi: 10.1128/jb.176.6.1711-1717.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Walmsley R. H. Temperature dependence of mating-pair formation in Escherichia coli. J Bacteriol. 1976 Apr;126(1):222–224. doi: 10.1128/jb.126.1.222-224.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ward J. E., Jr, Dale E. M., Binns A. N. Activity of the Agrobacterium T-DNA transfer machinery is affected by virB gene products. Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):9350–9354. doi: 10.1073/pnas.88.20.9350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ward J. E., Jr, Dale E. M., Christie P. J., Nester E. W., Binns A. N. Complementation analysis of Agrobacterium tumefaciens Ti plasmid virB genes by use of a vir promoter expression vector: virB9, virB10, and virB11 are essential virulence genes. J Bacteriol. 1990 Sep;172(9):5187–5199. doi: 10.1128/jb.172.9.5187-5199.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Willetts N., Crowther C. Mobilization of the non-conjugative IncQ plasmid RSF1010. Genet Res. 1981 Jun;37(3):311–316. doi: 10.1017/s0016672300020310. [DOI] [PubMed] [Google Scholar]
- Winans S. C., Allenza P., Stachel S. E., McBride K. E., Nester E. W. Characterization of the virE operon of the Agrobacterium Ti plasmid pTiA6. Nucleic Acids Res. 1987 Jan 26;15(2):825–837. doi: 10.1093/nar/15.2.825. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zhang L. H., Kerr A. A diffusible compound can enhance conjugal transfer of the Ti plasmid in Agrobacterium tumefaciens. J Bacteriol. 1991 Mar;173(6):1867–1872. doi: 10.1128/jb.173.6.1867-1872.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zhang L., Murphy P. J., Kerr A., Tate M. E. Agrobacterium conjugation and gene regulation by N-acyl-L-homoserine lactones. Nature. 1993 Apr 1;362(6419):446–448. doi: 10.1038/362446a0. [DOI] [PubMed] [Google Scholar]
- Zupan J. R., Zambryski P. Transfer of T-DNA from Agrobacterium to the plant cell. Plant Physiol. 1995 Apr;107(4):1041–1047. doi: 10.1104/pp.107.4.1041. [DOI] [PMC free article] [PubMed] [Google Scholar]