Skip to main content
The Plant Cell logoLink to The Plant Cell
. 1989 Nov;1(11):1057–1067. doi: 10.1105/tpc.1.11.1057

Functional expression of the leftward open reading frames of the A component of tomato golden mosaic virus in transgenic tobacco plants.

L Hanley-Bowdoin 1, J S Elmer 1, S G Rogers 1
PMCID: PMC159843  PMID: 2562559

Abstract

The genome of the geminivirus tomato golden mosaic virus (TGMV) consists of two circular DNA molecules designated as components A and B. We have constructed Nicotiana benthamiana plants that are transgenic for the three overlapping open reading frames, AL1, AL2, and AL3, from the left side of TGMV A. In the transgenic plants, the AL open reading frames are under the control of the cauliflower mosaic virus (CaMV) 35S promoter. In TGMV infectivity assays, seven of 10 transgenic lines complemented TGMV A variants with mutations in AL1, AL2, or AL3 when co-inoculated with the B component. The 35S-AL construct was transcribed as a single RNA species in the transgenic plants, indicating that AL1, AL2, and AL3 were expressed from a polycistronic mRNA. This differs from the complex transcription pattern in TGMV-infected plants, which contains five AL transcripts. There was no quantitative correlation between the efficiency of complementation in the infectivity assay and the level of expression of transgenic AL RNA in the leaves of a transgenic line. One line that failed to complement defects in AL1, AL2, and AL3 in infectivity assays contained high levels of transgenic AL RNA and functional AL1 protein. These results provide evidence that chromosomal position can affect the cell- and tissue-specific transcription of the 35S promoter in transgenic plants. Comparison of the complementing plants and wild-type infected plants may provide insight into the TGMV infection process and the use of the CaMV 35S promoter for gene expression in transgenic plants.

Full Text

The Full Text of this article is available as a PDF (1.9 MB).

Selected References

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

  1. Accotto G. P., Donson J., Mullineaux P. M. Mapping of Digitaria streak virus transcripts reveals different RNA species from the same transcription unit. EMBO J. 1989 Apr;8(4):1033–1039. doi: 10.1002/j.1460-2075.1989.tb03470.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baughman G. A., Jacobs J. D., Howell S. H. Cauliflower mosaic virus gene VI produces a symptomatic phenotype in transgenic tobacco plants. Proc Natl Acad Sci U S A. 1988 Feb;85(3):733–737. doi: 10.1073/pnas.85.3.733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benfey P. N., Chua N. H. Regulated genes in transgenic plants. Science. 1989 Apr 14;244(4901):174–181. doi: 10.1126/science.244.4901.174. [DOI] [PubMed] [Google Scholar]
  4. Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
  5. Chang L. J., Pryciak P., Ganem D., Varmus H. E. Biosynthesis of the reverse transcriptase of hepatitis B viruses involves de novo translational initiation not ribosomal frameshifting. Nature. 1989 Jan 26;337(6205):364–368. doi: 10.1038/337364a0. [DOI] [PubMed] [Google Scholar]
  6. Day M. L., Schwartz D., Wiegand R. C., Stockman P. T., Brunnert S. R., Tolunay H. E., Currie M. G., Standaert D. G., Needleman P. Ventricular atriopeptin. Unmasking of messenger RNA and peptide synthesis by hypertrophy or dexamethasone. Hypertension. 1987 May;9(5):485–491. doi: 10.1161/01.hyp.9.5.485. [DOI] [PubMed] [Google Scholar]
  7. Deom C. M., Oliver M. J., Beachy R. N. The 30-kilodalton gene product of tobacco mosaic virus potentiates virus movement. Science. 1987 Jul 24;237(4813):389–394. doi: 10.1126/science.237.4813.389. [DOI] [PubMed] [Google Scholar]
  8. Elmer J. S., Brand L., Sunter G., Gardiner W. E., Bisaro D. M., Rogers S. G. Genetic analysis of the tomato golden mosaic virus. II. The product of the AL1 coding sequence is required for replication. Nucleic Acids Res. 1988 Jul 25;16(14B):7043–7060. doi: 10.1093/nar/16.14.7043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Etessami P., Callis R., Ellwood S., Stanley J. Delimitation of essential genes of cassava latent virus DNA 2. Nucleic Acids Res. 1988 Jun 10;16(11):4811–4829. doi: 10.1093/nar/16.11.4811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fang R. X., Nagy F., Sivasubramaniam S., Chua N. H. Multiple cis regulatory elements for maximal expression of the cauliflower mosaic virus 35S promoter in transgenic plants. Plant Cell. 1989 Jan;1(1):141–150. doi: 10.1105/tpc.1.1.141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gardiner W. E., Sunter G., Brand L., Elmer J. S., Rogers S. G., Bisaro D. M. Genetic analysis of tomato golden mosaic virus: the coat protein is not required for systemic spread or symptom development. EMBO J. 1988 Apr;7(4):899–904. doi: 10.1002/j.1460-2075.1988.tb02894.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hamilton W. D., Bisaro D. M., Coutts R. H., Buck K. W. Demonstration of the bipartite nature of the genome of a single-stranded DNA plant virus by infection with the cloned DNA components. Nucleic Acids Res. 1983 Nov 11;11(21):7387–7396. doi: 10.1093/nar/11.21.7387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hamilton W. D., Stein V. E., Coutts R. H., Buck K. W. Complete nucleotide sequence of the infectious cloned DNA components of tomato golden mosaic virus: potential coding regions and regulatory sequences. EMBO J. 1984 Sep;3(9):2197–2205. doi: 10.1002/j.1460-2075.1984.tb02114.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hammer R. E., Krumlauf R., Camper S. A., Brinster R. L., Tilghman S. M. Diversity of alpha-fetoprotein gene expression in mice is generated by a combination of separate enhancer elements. Science. 1987 Jan 2;235(4784):53–58. doi: 10.1126/science.2432657. [DOI] [PubMed] [Google Scholar]
  15. Hanley-Bowdoin L., Elmer J. S., Rogers S. G. Transient expression of heterologous RNAs using tomato golden mosaic virus. Nucleic Acids Res. 1988 Nov 25;16(22):10511–10528. doi: 10.1093/nar/16.22.10511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hanley-Bowdoin L., Orozco E. M., Jr, Chua N. H. In vitro synthesis and processing of a maize chloroplast transcript encoded by the ribulose 1,5-bisphosphate carboxylase large subunit gene. Mol Cell Biol. 1985 Oct;5(10):2733–2745. doi: 10.1128/mcb.5.10.2733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Horsch R. B., Klee H. J. Rapid assay of foreign gene expression in leaf discs transformed by Agrobacterium tumefaciens: Role of T-DNA borders in the transfer process. Proc Natl Acad Sci U S A. 1986 Jun;83(12):4428–4432. doi: 10.1073/pnas.83.12.4428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hughes S., Mellstrom K., Kosik E., Tamanoi F., Brugge J. Mutation of a termination codon affects src initiation. Mol Cell Biol. 1984 Sep;4(9):1738–1746. doi: 10.1128/mcb.4.9.1738. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hyde-DeRuyscher R., Carmichael G. G. Polyomavirus early-late switch is not regulated at the level of transcription initiation and is associated with changes in RNA processing. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8993–8997. doi: 10.1073/pnas.85.23.8993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Jefferson R. A., Kavanagh T. A., Bevan M. W. GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 1987 Dec 20;6(13):3901–3907. doi: 10.1002/j.1460-2075.1987.tb02730.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Jones J. D., Dunsmuir P., Bedbrook J. High level expression of introduced chimaeric genes in regenerated transformed plants. EMBO J. 1985 Oct;4(10):2411–2418. doi: 10.1002/j.1460-2075.1985.tb03949.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Joshi C. P. An inspection of the domain between putative TATA box and translation start site in 79 plant genes. Nucleic Acids Res. 1987 Aug 25;15(16):6643–6653. doi: 10.1093/nar/15.16.6643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Khoury G., May E. Regulation of early and late simian virus 40 transcription: overproduction of early viral RNA in the absence of a functional T-antigen. J Virol. 1977 Jul;23(1):167–176. doi: 10.1128/jvi.23.1.167-176.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kozak M. Bifunctional messenger RNAs in eukaryotes. Cell. 1986 Nov 21;47(4):481–483. doi: 10.1016/0092-8674(86)90609-4. [DOI] [PubMed] [Google Scholar]
  25. Lazarowitz S. G. Infectivity and complete nucleotide sequence of the genome of a South African isolate of maize streak virus. Nucleic Acids Res. 1988 Jan 11;16(1):229–249. doi: 10.1093/nar/16.1.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Morris-Krsinich B. A., Mullineaux P. M., Donson J., Boulton M. I., Markham P. G., Short M. N., Davies J. W. Bidirectional transcription of maize streak virus DNA and identification of the coat protein gene. Nucleic Acids Res. 1985 Oct 25;13(20):7237–7256. doi: 10.1093/nar/13.20.7237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Nagy F., Kay S. A., Boutry M., Hsu M. Y., Chua N. H. Phytochrome-controlled expression of a wheat Cab gene in transgenic tobacco seedlings. EMBO J. 1986 Jun;5(6):1119–1124. doi: 10.1002/j.1460-2075.1986.tb04335.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nagy F., Morelli G., Fraley R. T., Rogers S. G., Chua N. H. Photoregulated expression of a pea rbcS gene in leaves of transgenic plants. EMBO J. 1985 Dec 1;4(12):3063–3068. doi: 10.1002/j.1460-2075.1985.tb04046.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Nevins J. R., Ginsberg H. S., Blanchard J. M., Wilson M. C., Darnell J. E., Jr Regulation of the primary expression of the early adenovirus transcription units. J Virol. 1979 Dec;32(3):727–733. doi: 10.1128/jvi.32.3.727-733.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nomiyama H., Fromental C., Xiao J. H., Chambon P. Cell-specific activity of the constituent elements of the simian virus 40 enhancer. Proc Natl Acad Sci U S A. 1987 Nov;84(22):7881–7885. doi: 10.1073/pnas.84.22.7881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ondek B., Shepard A., Herr W. Discrete elements within the SV40 enhancer region display different cell-specific enhancer activities. EMBO J. 1987 Apr;6(4):1017–1025. doi: 10.1002/j.1460-2075.1987.tb04854.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Peabody D. S., Subramani S., Berg P. Effect of upstream reading frames on translation efficiency in simian virus 40 recombinants. Mol Cell Biol. 1986 Jul;6(7):2704–2711. doi: 10.1128/mcb.6.7.2704. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rogers S. G., Bisaro D. M., Horsch R. B., Fraley R. T., Hoffmann N. L., Brand L., Elmer J. S., Lloyd A. M. Tomato golden mosaic virus A component DNA replicates autonomously in transgenic plants. Cell. 1986 May 23;45(4):593–600. doi: 10.1016/0092-8674(86)90291-6. [DOI] [PubMed] [Google Scholar]
  34. Sample J., Hummel M., Braun D., Birkenbach M., Kieff E. Nucleotide sequences of mRNAs encoding Epstein-Barr virus nuclear proteins: a probable transcriptional initiation site. Proc Natl Acad Sci U S A. 1986 Jul;83(14):5096–5100. doi: 10.1073/pnas.83.14.5096. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Schalk H. J., Matzeit V., Schiller B., Schell J., Gronenborn B. Wheat dwarf virus, a geminivirus of graminaceous plants needs splicing for replication. EMBO J. 1989 Feb;8(2):359–364. doi: 10.1002/j.1460-2075.1989.tb03385.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Schirm S., Jiricny J., Schaffner W. The SV40 enhancer can be dissected into multiple segments, each with a different cell type specificity. Genes Dev. 1987 Mar;1(1):65–74. doi: 10.1101/gad.1.1.65. [DOI] [PubMed] [Google Scholar]
  37. Sunter G., Gardiner W. E., Bisaro D. M. Identification of tomato golden mosaic virus-specific RNAs in infected plants. Virology. 1989 May;170(1):243–250. doi: 10.1016/0042-6822(89)90372-3. [DOI] [PubMed] [Google Scholar]
  38. Townsend R., Stanley J., Curson S. J., Short M. N. Major polyadenylated transcripts of cassava latent virus and location of the gene encoding coat protein. EMBO J. 1985 Jan;4(1):33–37. doi: 10.1002/j.1460-2075.1985.tb02313.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. de Martynoff G., Pays E., Vassart G. Synthesis of a full length DNA complementary to thyroglobulin 33 S messenger RNA. Biochem Biophys Res Commun. 1980 Apr 14;93(3):645–653. doi: 10.1016/0006-291x(80)91127-4. [DOI] [PubMed] [Google Scholar]

Articles from The Plant Cell are provided here courtesy of Oxford University Press

RESOURCES