Abstract
By analysing mRNAs with the polymerase chain reaction (PCR) and by studying in vitro generated mutants we have identified an intron in the genome of wheat dwarf virus (WDV), a geminivirus of cereals. Polypeptides whose expression is essential for the replication of the viral DNA have been defined. They are encoded by two distinct overlapping open reading frames (ORFs). The joining of these two ORFs by deletion of the intron as well as the introduction of a frameshift mutation within the intron do not prevent replication of the viral genome in suspension culture cells. In contrast to WDV, the geminiviruses of dicotyledonous plants possess a single continuous ORF, highly homologous to the two individual ones of WDV. We propose that mRNA splicing is a common feature of all geminiviruses of the Gramineae and might contribute to their host class specificity. The existence of a functional intron is a novel finding for the plant viruses.
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Selected References
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- Andersen M. T., Richardson K. A., Harbison S. A., Morris B. A. Nucleotide sequence of the geminivirus chloris striate mosaic virus. Virology. 1988 Jun;164(2):443–449. doi: 10.1016/0042-6822(88)90558-2. [DOI] [PubMed] [Google Scholar]
- Donson J., Accotto G. P., Boulton M. I., Mullineaux P. M., Davies J. W. The nucleotide sequence of a geminivirus from Digitaria sanguinalis. Virology. 1987 Nov;161(1):160–169. doi: 10.1016/0042-6822(87)90182-6. [DOI] [PubMed] [Google Scholar]
- Donson J., Morris-Krsinich B. A., Mullineaux P. M., Boulton M. I., Davies J. W. A putative primer for second-strand DNA synthesis of maize streak virus is virion-associated. EMBO J. 1984 Dec 20;3(13):3069–3073. doi: 10.1002/j.1460-2075.1984.tb02260.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gay N. J., Walker J. E. Homology between human bladder carcinoma oncogene product and mitochondrial ATP-synthase. Nature. 1983 Jan 20;301(5897):262–264. doi: 10.1038/301262a0. [DOI] [PubMed] [Google Scholar]
- Green M. R. Pre-mRNA splicing. Annu Rev Genet. 1986;20:671–708. doi: 10.1146/annurev.ge.20.120186.003323. [DOI] [PubMed] [Google Scholar]
- 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]
- Howarth A. J., Caton J., Bossert M., Goodman R. M. Nucleotide sequence of bean golden mosaic virus and a model for gene regulation in geminiviruses. Proc Natl Acad Sci U S A. 1985 Jun;82(11):3572–3576. doi: 10.1073/pnas.82.11.3572. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Howell S. H. Physical structure and genetic organisation of the genome of maize streak virus (Kenyan isolate). Nucleic Acids Res. 1984 Oct 11;12(19):7359–7375. doi: 10.1093/nar/12.19.7359. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kramer W., Drutsa V., Jansen H. W., Kramer B., Pflugfelder M., Fritz H. J. The gapped duplex DNA approach to oligonucleotide-directed mutation construction. Nucleic Acids Res. 1984 Dec 21;12(24):9441–9456. doi: 10.1093/nar/12.24.9441. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- MacDowell S. W., Macdonald H., Hamilton W. D., Coutts R. H., Buck K. W. The nucleotide sequence of cloned wheat dwarf virus DNA. EMBO J. 1985 Sep;4(9):2173–2180. doi: 10.1002/j.1460-2075.1985.tb03912.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
- Saiki R. K., Scharf S., Faloona F., Mullis K. B., Horn G. T., Erlich H. A., Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science. 1985 Dec 20;230(4732):1350–1354. doi: 10.1126/science.2999980. [DOI] [PubMed] [Google Scholar]
- Stanley J., Markham P. G., Callis R. J., Pinner M. S. The nucleotide sequence of an infectious clone of the geminivirus beet curly top virus. EMBO J. 1986 Aug;5(8):1761–1767. doi: 10.1002/j.1460-2075.1986.tb04424.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stanley J. The molecular biology of geminiviruses. Adv Virus Res. 1985;30:139–177. doi: 10.1016/s0065-3527(08)60450-9. [DOI] [PubMed] [Google Scholar]
- 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]
- Townsend R., Watts J., Stanley J. Synthesis of viral DNA forms in Nicotiana plumbaginifolia protoplasts inoculated with cassava latent virus (CLV); evidence for the independent replication of one component of the CLV genome. Nucleic Acids Res. 1986 Feb 11;14(3):1253–1265. doi: 10.1093/nar/14.3.1253. [DOI] [PMC free article] [PubMed] [Google Scholar]