Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1993 Nov;67(11):6432–6438. doi: 10.1128/jvi.67.11.6432-6438.1993

Two amino acid substitutions in the tomato mosaic virus 30-kilodalton movement protein confer the ability to overcome the Tm-2(2) resistance gene in the tomato.

H Weber 1, S Schultze 1, A J Pfitzner 1
PMCID: PMC238078  PMID: 8411345

Abstract

The Tm-2(2) resistance gene is used in most commercial tomato cultivars for protection against infection with tobacco mosaic virus and its close relative tomato mosaic virus (ToMV). To study the mechanism of this resistance gene, cDNA clones encompassing the complete genome of a ToMV strain (ToMV-2(2)) that was able to break the Tm-2(2) resistance were generated. Chimeric full-length viral cDNA clones were constructed under the control of the cauliflower mosaic virus 35S RNA promoter, combining parts of the wild-type virus and ToMV-2(2). Using these clones in cDNA infection experiments, we showed that the 30-kDa movement protein of ToMV-2(2) is responsible for overcoming the Tm-2(2) resistance gene in the tomato. DNA sequence analysis revealed four amino acid exchanges between the 30-kDa proteins from wild-type ToMV and ToMV-2(2), Lys-130 to Glu, Gly-184 to Glu, Ser-238 to Arg, and Lys-244 to Glu. To clarify the involvement of the altered amino acid residues in the resistance-breaking properties of the ToMV-2(2) movement protein, different combinations of these amino acid exchanges were introduced in the genome of wild-type ToMV. Only one mutant strain which contained two amino acid substitutions, Arg-238 and Glu-244, was able to multiply in Tm-2(2) tomato plants. Both amino acid exchanges are found within the carboxy-terminal region of the movement protein, which displays a high variability among different tobamoviruses and has been shown to be dispensable for virus transport in tobacco plants. These observations suggest that the resistance conferred by the Tm-2(2) gene against ToMV depends on specific recognition events in this host-pathogen interaction rather than interfering with fundamental functions of the 30-kDa protein.

Full text

PDF
6432

Images in this article

Selected References

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

  1. Calder V. L., Palukaitis P. Nucleotide sequence analysis of the movement genes of resistance breaking strains of tomato mosaic virus. J Gen Virol. 1992 Jan;73(Pt 1):165–168. doi: 10.1099/0022-1317-73-1-165. [DOI] [PubMed] [Google Scholar]
  2. Gafny R., Lapidot M., Berna A., Holt C. A., Deom C. M., Beachy R. N. Effects of terminal deletion mutations on function of the movement protein of tobacco mosaic virus. Virology. 1992 Apr;187(2):499–507. doi: 10.1016/0042-6822(92)90452-u. [DOI] [PubMed] [Google Scholar]
  3. Goelet P., Lomonossoff G. P., Butler P. J., Akam M. E., Gait M. J., Karn J. Nucleotide sequence of tobacco mosaic virus RNA. Proc Natl Acad Sci U S A. 1982 Oct;79(19):5818–5822. doi: 10.1073/pnas.79.19.5818. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Meshi T., Ishikawa M., Motoyoshi F., Semba K., Okada Y. In vitro transcription of infectious RNAs from full-length cDNAs of tobacco mosaic virus. Proc Natl Acad Sci U S A. 1986 Jul;83(14):5043–5047. doi: 10.1073/pnas.83.14.5043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Meshi T., Motoyoshi F., Maeda T., Yoshiwoka S., Watanabe H., Okada Y. Mutations in the tobacco mosaic virus 30-kD protein gene overcome Tm-2 resistance in tomato. Plant Cell. 1989 May;1(5):515–522. doi: 10.1105/tpc.1.5.515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ohno T., Aoyagi M., Yamanashi Y., Saito H., Ikawa S., Meshi T., Okada Y. Nucleotide sequence of the tobacco mosaic virus (tomato strain) genome and comparison with the common strain genome. J Biochem. 1984 Dec;96(6):1915–1923. doi: 10.1093/oxfordjournals.jbchem.a135026. [DOI] [PubMed] [Google Scholar]
  8. Pfitzner U. M., Goodman H. M. Isolation and characterization of cDNA clones encoding pathogenesis-related proteins from tobacco mosaic virus infected tobacco plants. Nucleic Acids Res. 1987 Jun 11;15(11):4449–4465. doi: 10.1093/nar/15.11.4449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Pfitzner U. M., Pfitzner A. J. Expression of a viral avirulence gene in transgenic plants is sufficient to induce the hypersensitive defense reaction. Mol Plant Microbe Interact. 1992 Jul-Aug;5(4):318–321. doi: 10.1094/mpmi-5-318. [DOI] [PubMed] [Google Scholar]
  10. Saito T., Imai Y., Meshi T., Okada Y. Interviral homologies of the 30K proteins of tobamoviruses. Virology. 1988 Dec;167(2):653–656. [PubMed] [Google Scholar]
  11. 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]
  12. Watanabe Y., Kishibayashi N., Motoyoshi F., Okada Y. Characterization of Tm-1 gene action on replication of common isolates and a resistance-breaking isolate of TMV. Virology. 1987 Dec;161(2):527–532. doi: 10.1016/0042-6822(87)90147-4. [DOI] [PubMed] [Google Scholar]
  13. Watanabe Y., Ogawa T., Okada Y. In vivo phosphorylation of the 30-kDa protein of tobacco mosaic virus. FEBS Lett. 1992 Nov 23;313(2):181–184. doi: 10.1016/0014-5793(92)81440-w. [DOI] [PubMed] [Google Scholar]
  14. Weber H., Haeckel P., Pfitzner A. J. A cDNA clone of tomato mosaic virus is infectious in plants. J Virol. 1992 Jun;66(6):3909–3912. doi: 10.1128/jvi.66.6.3909-3912.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES