1887

Abstract

The tomato golden mosaic virus (TGMV) coat protein and AL1 genes are located in opposite directions on either side of an intergenic region. To enable the effects of the AL1, AL2 and AL3 gene products on expression of the coat protein and AL1 genes to be studied simultaneously, a plasmid was constructed, containing the intergenic region linked on one side to a 5′ -terminal portion of the AL1 gene fused to a -glucuronidase (GUS) reporter gene (to replace most of the AL1 gene) and on the other side to a neomycin phosphotransferase (NEO) reporter gene (to replace the coat protein gene). This GUS-NEO plasmid was mixed with plant expression plasmids containing the AL1, AL2 or AL3 coding regions, the DNA was transformed into protoplasts and GUS activities and NEO protein levels were measured. Control transformations were carried out with the GUS-NEO plasmid mixed with the AL1, AL2 or AL3 plasmids in which mutations were introduced to prevent translation of the open reading frames (ORFs). The results showed that transactivation of the coat protein gene by the AL2 gene product and suppression of the AL1 gene by the expression of AL1 DNA (both reported previously) can occur simultaneously. It was also shown that expression of AL4, a small ORF contained within AL1 DNA but in a different reading frame, as well as expression of ORF AL1, can cause significant suppression of AL1 gene expression. Neither the AL1 nor the AL3 gene products affected the expression of the coat protein gene.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-75-4-721
1994-04-01
2021-09-24
Loading full text...

Full text loading...

/deliver/fulltext/jgv/75/4/JV0750040721.html?itemId=/content/journal/jgv/10.1099/0022-1317-75-4-721&mimeType=html&fmt=ahah

References

  1. Brough C. L., Haves R. J., Morgan A. J., Coutts R. H. A., Buck K. W. 1988; Effects of mutagenesis in vitro on the ability of cloned tomato golden mosaic virus DNA to infect Nicotiana benthamiana plants. Journal of General Virology 69:503–514
    [Google Scholar]
  2. Brough C. L., Sunter G., Gardiner W. E., Bisaro D. M. 1992; Kinetics of tomato golden mosaic virus DNA replication and coat protein promoter activity in Nicotiana tabacum protoplasts. Virology 187:1–9
    [Google Scholar]
  3. Dente L., Sollazo M., Baldari C., Cesarini G., Cortese R. 1985; The pEMBL family of single-stranded vectors. In DNA Cloning. A Practical Approach 1 pp 101–107 Glover D. M. Edited by Oxford: IRL Press;
    [Google Scholar]
  4. Elmer J. S., Brand L., Sunter G., Gardiner W. E., Bisaro D. M., Rogers S. G. 1988; Genetic analysis of the tomato golden mosaic virus II. The product of the ALI coding sequence is required for replication. Nucleic Acids Research 16:7043–7060
    [Google Scholar]
  5. Fisher R. A. 1954 Statistical Methods for Research Workers, 12th edn. London: Oliver & Boyd;
    [Google Scholar]
  6. Fisher R. A., Yates F. 1963 Statistical Tables for Biological, Agricultural and Medical Research, 6th edn. Edinburgh: Oliver & Boyd;
    [Google Scholar]
  7. Fontes E. P. B., Luckow V. A., Hanley-Bowdoin L. 1992; A geminivirus replication protein is a sequence-specific DNA binding protein. Plant Cell 4:597–608
    [Google Scholar]
  8. Francki R. I. B., Fauquet C. M., Knudson D. L., Brown F. editor 1991; Classification and Nomenclature of Viruses. Fifth Report of the International Committee on Taxonomy of Viruses. Archives of Virology, supplementum 2:173–177
    [Google Scholar]
  9. Gallie D. R., Sleat D. E., Watts J. W., Turner P. C., Wilson T. M. A. 1987; Characterisation of the 5′-leader sequence of tobacco mosaic virus RNA as a general enhancer of translation in vitro. Gene 217:217–225
    [Google Scholar]
  10. Haley A., Zhan X., Richardson K., Head K., Morris B. 1992; Regulation of the activities of African cassava mosaic virus promoters by the AC1, AC2 and AC3 gene products. Virology 188:905–909
    [Google Scholar]
  11. Hamilton W. D. O., Bisaro D. M., Coutts R. H. A., Buck K. W. 1983; Demonstration of the bipartite nature of the genome of a single-stranded DNA plant virus by infection with the cloned DNA components. Nucleic Acids Research 11:7387–7396
    [Google Scholar]
  12. Hamilton W. D. O., Stein V. E., Coutts R. H. A., Buck K. W. 1984; Complete nucleotide sequence of the infectious cloned DNA components of tomato golden mosaic virus: potential coding regions and regulatory sequences. EMBO Journal 3:2197–2205
    [Google Scholar]
  13. Hayes R. J., Buck K. W. 1989; Replication of tomato golden mosaic virus DNA m transgenic plants expressing open reading frames (ORFs) of DNA A: requirement of ORF AL2 for production of single-stranded DNA. Nucleic Acids Research 17:10213–10222
    [Google Scholar]
  14. Jefferson R. A. 1987; Assaying chimeric genes in plants: the GUS gene fusion system. Plant Molecular Biology Reporter 5:387–405
    [Google Scholar]
  15. Jefferson R. A., Kavanagh T. A., Bevan M W. 1987; GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO Journal 6:3901–3907
    [Google Scholar]
  16. Kallender H., Petty I. T. D., Stein V. E., Panico M., Blench I. P., Etienne A. T., Morris H. R., Coutts R. H. A., Buck K. W. 1988; Identification of the coat protein gene of tomato golden mosaic virus. Journal of General Virology 69:1351–1357
    [Google Scholar]
  17. Kay R., Chan A., Daly M., Mcpherson J. 1987; Duplication of CaMV 35S promoter sequences creates a strong enhancer for plant genes. Science 236:1299–1302
    [Google Scholar]
  18. Künkel T. A., Roberts J. O., Zakour R. A. 1987; Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods in Enzymology 154:367–382
    [Google Scholar]
  19. Lütcke H. A., Chow K. C., Mickel F. S., Moss K. A., Kern H. F., Scheele G. A. 1987; Selection of AUG initiation codons differs in plants and animals. EMBO Journal 6:43–48
    [Google Scholar]
  20. Menczel L., Nagy F., Kiss Z., Maliga P. 1981; Streptomycin- resistant and sensitive somatic hybrids of N. tabacum + N. knitiana: correlation of resistance to N. tabacum plastids. Theoretical and Applied Genetics 59:191–195
    [Google Scholar]
  21. Negrutio I., Shillito R., Potrykus I., Biasini G., Sala F. 1987; Hybrid genes in the analysis of transformation conditions. I. Setting up a simple method for direct gene transfer in plant protoplasts. Plant Molecular Biology 8:363–373
    [Google Scholar]
  22. Odell J. T., Nagy F., Chua N.-H. 1985; Identification of DNA sequences required for the activity of the cauliflower mosaic virus 35S promoter. Nature; London: 313810–812
    [Google Scholar]
  23. Petty I. T. D., Coutts R. H. A., Buck K. W. 1986; Geminivirus coat protein promoter sequences can function m Escherichia coli. Nucleic Acids Research 14:5113
    [Google Scholar]
  24. Power J. B., Chapman J. V. 1985; Isolation, culture and genetic manipulation of plant protoplasts. In Plant Cell Culture o A Practical Approach pp 37–66 Dixon R. A. Edited by Oxford: IRL Press;
    [Google Scholar]
  25. Rogers S. G., Bisaro D. M., Horsch R. B., Fraley R. T., Hoffman N. L., Brand L., Elmer J. S., Lloyd A. M. 1986; Tomato golden mosaic virus A component DNA replicates autonomously in transgenic plants. Cell 45:593–600
    [Google Scholar]
  26. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. 1988; Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491
    [Google Scholar]
  27. Saunders K., Lucy A., Stanley J. 1991; DNA forms of the geminivirus African cassava latent virus consistent with a rolling circle mechanism of replication. Nucleic Acids Research 19:2325–2330
    [Google Scholar]
  28. Stanley J., Latham J. R. 1992; A symptom variant of beet curly top geminivirus produced by mutation of open reading frame C4. Virology 190:506–509
    [Google Scholar]
  29. Stanley J., Latham J. R., Pinner M. S., Bedford I., Markham P. G. 1992; Mutational analysis of the monopartite geminivirus beet curly top virus. Virology 191:396–405
    [Google Scholar]
  30. Stenger D. C., Revrngton G. N., Stevenson M. C., Bisaro D. M. 1991; Replicational release of geminivirus genomes from tandemly repeated copies: evidence for rolling circle replication of plant viral DNA. Proceedings of the National Academv of Sciences U.S.A.: 888029–8033
    [Google Scholar]
  31. Sunter G., Bisaro D M. 1989; Transcriptional map of the B genome component of tomato golden mosaic virus and comparison with A component transcripts. Virology 173:647–655
    [Google Scholar]
  32. Sunter G., Bisaro D. M. 1991; Transactivation in geminivirus: AL2 gene product is needed for coat protein expression. Virology 180:416–419
    [Google Scholar]
  33. Sunter G., Bisaro D. M. 1992; Transactivation of geminivirus AR1 and BR1 gene expression by the viral AL2 gene product occurs at the level of transcription. Plant Cell 4:1321–1331
    [Google Scholar]
  34. Sunter G., Hartitz M. D., Hormuzdi S. G., Brough C. L., Bisaro D. M. 1990; Genetic analysis of tomato golden mosaic virus: ORF AL2 is required for coat protein accumulation while ORF AL3 is necessary for efficient DNA replication. Virology 179:69–77
    [Google Scholar]
  35. Sunter G., Hartitz M. D., Bisaro D. M. 1993; Tomato golden mosaic virus leftward gene expression: autoregulation of geminivirus replication protein. Virology 195:275–280
    [Google Scholar]
  36. Tan M. M. C., Boerrigter H. S., Kool A. J. 1987; A rapid procedure for plant regeneration from protoplasts isolated from suspension cultures and leaf mesophyll cells of wild Solanum species and Lycopersicon pennelii. Plant Science 49:63–72
    [Google Scholar]
  37. Thömmes P., Osman T. A. M., Hayes R. J., Buck K. W. 1993; TGMV replication protein AL1 binds preferentially to singlestranded DNA of the common region. FEBS Letters 319:95–99
    [Google Scholar]
  38. Yamaya J., Yoshioka M., Meshi T., Okada Y., Ohno T. 1988; Expression of tobacco mosaic virus RNA in transgenic plants. Molecular and General Genetics 211:520–525
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-75-4-721
Loading
/content/journal/jgv/10.1099/0022-1317-75-4-721
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error