Skip to main content
PMC home page
The Plant Cell logoLink to The Plant Cell
. 1992 Dec;4(12):1471–1484. doi: 10.1105/tpc.4.12.1471

Cauliflower Mosaic Virus Gene VI Controls Translation from Dicistronic Expression Units in Transgenic Arabidopsis Plants.

C Zijlstra 1, T Hohn 1
PMCID: PMC160234  PMID: 12297640

Abstract

Transformed Arabidopsis plants were used to study the effect of the cauliflower mosaic virus (CaMV) inclusion body protein on translation of dicistronic RNA. Reporter plants contain a dicistronic transcription unit with CaMV open reading frame VII (ORF VII) as the first and the [beta]-glucuronidase (GUS) reporter ORF as the second cistron. "Transactivator plants" contain CaMV ORF VI under the control of the strong CaMV 35S promoter. The transactivator plants were difficult to regenerate and showed an abnormal phenotype. Expression of GUS activity in the reporter plants was very low, but high GUS activity could be induced by introduction of gene VI, either by crossing with plants containing gene VI as a transgene or by infection with CaMV. Histological GUS assays showed that transactivation occurred in all types of tissue and at all developmental stages. The practical implications of the induction of GUS expression from the dicistronic unit by virus infection are discussed.

Full Text

The Full Text of this article is available as a PDF (3.2 MB).

Selected References

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

  1. 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]
  2. Benfey P. N., Chua N. H. The Cauliflower Mosaic Virus 35S Promoter: Combinatorial Regulation of Transcription in Plants. Science. 1990 Nov 16;250(4983):959–966. doi: 10.1126/science.250.4983.959. [DOI] [PubMed] [Google Scholar]
  3. Bevan M. Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res. 1984 Nov 26;12(22):8711–8721. doi: 10.1093/nar/12.22.8711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bonneville J. M., Volovitch M., Modjtahedi N., Demery D., Yot P. In vitro synthesis of cauliflower mosaic virus DNA in viroplasms. Adv Exp Med Biol. 1984;179:113–119. doi: 10.1007/978-1-4684-8730-5_11. [DOI] [PubMed] [Google Scholar]
  5. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  6. Chambers S. P., Prior S. E., Barstow D. A., Minton N. P. The pMTL nic- cloning vectors. I. Improved pUC polylinker regions to facilitate the use of sonicated DNA for nucleotide sequencing. Gene. 1988 Aug 15;68(1):139–149. doi: 10.1016/0378-1119(88)90606-3. [DOI] [PubMed] [Google Scholar]
  7. Citovsky V., Knorr D., Zambryski P. Gene I, a potential cell-to-cell movement locus of cauliflower mosaic virus, encodes an RNA-binding protein. Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2476–2480. doi: 10.1073/pnas.88.6.2476. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Damm B., Schmidt R., Willmitzer L. Efficient transformation of Arabidopsis thaliana using direct gene transfer to protoplasts. Mol Gen Genet. 1989 May;217(1):6–12. doi: 10.1007/BF00330935. [DOI] [PubMed] [Google Scholar]
  9. Daubert S. D., Schoelz J., Debao L., Shepherd R. J. Expression of disease symptoms in cauliflower mosaic virus genomic hybrids. J Mol Appl Genet. 1984;2(6):537–547. [PubMed] [Google Scholar]
  10. Daubert S., Routh G. Point mutations in cauliflower mosaic virus gene VI confer host-specific symptom changes. Mol Plant Microbe Interact. 1990 Sep-Oct;3(5):341–345. doi: 10.1094/mpmi-3-341. [DOI] [PubMed] [Google Scholar]
  11. De Zoeten G. A., Penswick J. R., Horisberger M. A., Ahl P., Schultze M., Hohn T. The expression, localization, and effect of a human interferon in plants. Virology. 1989 Sep;172(1):213–222. doi: 10.1016/0042-6822(89)90123-2. [DOI] [PubMed] [Google Scholar]
  12. Deblaere R., Bytebier B., De Greve H., Deboeck F., Schell J., Van Montagu M., Leemans J. Efficient octopine Ti plasmid-derived vectors for Agrobacterium-mediated gene transfer to plants. Nucleic Acids Res. 1985 Jul 11;13(13):4777–4788. doi: 10.1093/nar/13.13.4777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dhaese P., De Greve H., Decraemer H., Schell J., Van Montagu M. Rapid mapping of transposon insertion and deletion mutations in the large Ti-plasmids of Agrobacterium tumefaciens. Nucleic Acids Res. 1979 Dec 11;7(7):1837–1849. doi: 10.1093/nar/7.7.1837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Dixon L. K., Hohn T. Initiation of translation of the cauliflower mosaic virus genome from a polycistronic mRNA: evidence from deletion mutagenesis. EMBO J. 1984 Dec 1;3(12):2731–2736. doi: 10.1002/j.1460-2075.1984.tb02203.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  16. Fütterer J., Gordon K., Pfeiffer P., Sanfaçon H., Pisan B., Bonneville J. M., Hohn T. Differential inhibition of downstream gene expression by the cauliflower mosaic virus 35S RNA leader. Virus Genes. 1989 Sep;3(1):45–55. doi: 10.1007/BF00301986. [DOI] [PubMed] [Google Scholar]
  17. Fütterer J., Hohn T. Role of an upstream open reading frame in the translation of polycistronic mRNAs in plant cells. Nucleic Acids Res. 1992 Aug 11;20(15):3851–3857. doi: 10.1093/nar/20.15.3851. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gordon K., Pfeiffer P., Fütterer J., Hohn T. In vitro expression of cauliflower mosaic virus genes. EMBO J. 1988 Feb;7(2):309–317. doi: 10.1002/j.1460-2075.1988.tb02814.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gowda S., Wu F. C., Scholthof H. B., Shepherd R. J. Gene VI of figwort mosaic virus (caulimovirus group) functions in posttranscriptional expression of genes on the full-length RNA transcript. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9203–9207. doi: 10.1073/pnas.86.23.9203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hartley R. W. Barnase and barstar: two small proteins to fold and fit together. Trends Biochem Sci. 1989 Nov;14(11):450–454. doi: 10.1016/0968-0004(89)90104-7. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Kozak M. The scanning model for translation: an update. J Cell Biol. 1989 Feb;108(2):229–241. doi: 10.1083/jcb.108.2.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Mason W. S., Taylor J. M., Hull R. Retroid virus genome replication. Adv Virus Res. 1987;32:35–96. doi: 10.1016/s0065-3527(08)60474-1. [DOI] [PubMed] [Google Scholar]
  25. Mattanovich D., Rüker F., Machado A. C., Laimer M., Regner F., Steinkellner H., Himmler G., Katinger H. Efficient transformation of Agrobacterium spp. by electroporation. Nucleic Acids Res. 1989 Aug 25;17(16):6747–6747. doi: 10.1093/nar/17.16.6747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mittelsten Scheid O., Paszkowski J., Potrykus I. Reversible inactivation of a transgene in Arabidopsis thaliana. Mol Gen Genet. 1991 Aug;228(1-2):104–112. doi: 10.1007/BF00282454. [DOI] [PubMed] [Google Scholar]
  27. Murray M. G., Thompson W. F. Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res. 1980 Oct 10;8(19):4321–4325. doi: 10.1093/nar/8.19.4321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Natsoulis G., Boeke J. D. New antiviral strategy using capsid-nuclease fusion proteins. Nature. 1991 Aug 15;352(6336):632–635. doi: 10.1038/352632a0. [DOI] [PubMed] [Google Scholar]
  29. Olszewski N., Hagen G., Guilfoyle T. J. A transcriptionally active, covalently closed minichromosome of cauliflower mosaic virus DNA isolated from infected turnip leaves. Cell. 1982 Jun;29(2):395–402. doi: 10.1016/0092-8674(82)90156-8. [DOI] [PubMed] [Google Scholar]
  30. Pfeiffer P., Hohn T. Involvement of reverse transcription in the replication of cauliflower mosaic virus: a detailed model and test of some aspects. Cell. 1983 Jul;33(3):781–789. doi: 10.1016/0092-8674(83)90020-x. [DOI] [PubMed] [Google Scholar]
  31. Pietrzak M., Shillito R. D., Hohn T., Potrykus I. Expression in plants of two bacterial antibiotic resistance genes after protoplast transformation with a new plant expression vector. Nucleic Acids Res. 1986 Jul 25;14(14):5857–5868. doi: 10.1093/nar/14.14.5857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sohn G., Sautter C. R-phycoerythrin as a fluorescent label for immunolocalization of bound atrazine residues. J Histochem Cytochem. 1991 Jul;39(7):921–926. doi: 10.1177/39.7.1865109. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Stirpe F., Barbieri L., Battelli M. G., Soria M., Lappi D. A. Ribosome-inactivating proteins from plants: present status and future prospects. Biotechnology (N Y) 1992 Apr;10(4):405–412. doi: 10.1038/nbt0492-405. [DOI] [PubMed] [Google Scholar]
  35. Stratford R., Covey S. N. Segregation of cauliflower mosaic virus symptom genetic determinants. Virology. 1989 Oct;172(2):451–459. doi: 10.1016/0042-6822(89)90187-6. [DOI] [PubMed] [Google Scholar]
  36. Thomas C. M., Hull R., Bryant J. A., Maule A. J. Isolation of a fraction from cauliflower mosaic virus-infected protoplasts which is active in the synthesis of (+) and (-) strand viral DNA and reverse transcription of primed RNA templates. Nucleic Acids Res. 1985 Jun 25;13(12):4557–4576. doi: 10.1093/nar/13.12.4557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Woolston C. J., Covey S. N., Penswick J. R., Davies J. W. Aphid transmission and a polypeptide are specified by a defined region of the cauliflower mosaic virus genome. Gene. 1983 Jul;23(1):15–23. doi: 10.1016/0378-1119(83)90212-3. [DOI] [PubMed] [Google Scholar]
  38. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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

RESOURCES