Abstract
To determine the sequences essential for viral multiplication in the 5' untranslated leader sequence of tobacco mosaic virus RNA, mutant TMV-L (a tomato strain) RNAs which carry several deletions in this 71-nucleotide sequence were constructed by an in vitro transcription system and their multiplication was analyzed by introducing mutant RNA into tobacco protoplasts by electroporation. Large deletions of the sequence from nucleotides 9 to 47 or 25 to 71 abolished viral multiplication; when about 10-nucleotide deletions were introduced throughout this 5' leader sequence, only deletion of the sequence from nucleotides 2 to 8 abolished detectable viral multiplication. This mutant RNA, however, directed the synthesis of the 130,000-molecular-weight protein in a rabbit reticulocyte lysate in vitro translation system, and consequently this 5'-proximal portion appears likely to be essential for replication.
Full text
PDF



Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ahlquist P., Janda M. cDNA cloning and in vitro transcription of the complete brome mosaic virus genome. Mol Cell Biol. 1984 Dec;4(12):2876–2882. doi: 10.1128/mcb.4.12.2876. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gallie D. R., Sleat D. E., Watts J. W., Turner P. C., Wilson T. M. In vivo uncoating and efficient expression of foreign mRNAs packaged in TMV-like particles. Science. 1987 May 29;236(4805):1122–1124. doi: 10.1126/science.3472350. [DOI] [PubMed] [Google Scholar]
- Gallie D. R., Sleat D. E., Watts J. W., Turner P. C., Wilson T. M. Mutational analysis of the tobacco mosaic virus 5'-leader for altered ability to enhance translation. Nucleic Acids Res. 1988 Feb 11;16(3):883–893. doi: 10.1093/nar/16.3.883. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gallie D. R., Sleat D. E., Watts J. W., Turner P. C., Wilson T. M. The 5'-leader sequence of tobacco mosaic virus RNA enhances the expression of foreign gene transcripts in vitro and in vivo. Nucleic Acids Res. 1987 Apr 24;15(8):3257–3273. doi: 10.1093/nar/15.8.3257. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Ishikawa M., Meshi T., Motoyoshi F., Takamatsu N., Okada Y. In vitro mutagenesis of the putative replicase genes of tobacco mosaic virus. Nucleic Acids Res. 1986 Nov 11;14(21):8291–8305. doi: 10.1093/nar/14.21.8291. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jonard G., Richards K., Mohier E., Gerlinger P. Nucleotide sequence at the 5' extremity of tobacco-mosaic-virus RNA. 2. The coding region (nucleotides 69-236). Eur J Biochem. 1978 Mar 15;84(2):521–531. doi: 10.1111/j.1432-1033.1978.tb12195.x. [DOI] [PubMed] [Google Scholar]
- Konarska M., Filipowicz W., Domdey H., Gross H. J. Binding of ribosomes to linear and circular forms of the 5'-terminal leader fragment of tobacco-mosaic-virus RNA. Eur J Biochem. 1981 Feb;114(2):221–227. doi: 10.1111/j.1432-1033.1981.tb05139.x. [DOI] [PubMed] [Google Scholar]
- Kukla B. A., Guilley H. A., Jonard G. X., Richards K. E., Mundry K. W. Characterization of long guanosine-free RNA sequences from the Dahlemense and U2 strains of tobacco mosaic virus. Eur J Biochem. 1979 Jul;98(1):61–66. doi: 10.1111/j.1432-1033.1979.tb13160.x. [DOI] [PubMed] [Google Scholar]
- Kunkel T. A., Roberts J. D., Zakour R. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987;154:367–382. doi: 10.1016/0076-6879(87)54085-x. [DOI] [PubMed] [Google Scholar]
- Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
- 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]
- Meshi T., Ishikawa M., Takamatsu N., Ohno T., Okada Y. The 5'-terminal sequence of TMV RNA. Question on the polymorphism found in vulgare strain. FEBS Lett. 1983 Oct 17;162(2):282–285. doi: 10.1016/0014-5793(83)80772-8. [DOI] [PubMed] [Google Scholar]
- Meshi T., Motoyoshi F., Adachi A., Watanabe Y., Takamatsu N., Okada Y. Two concomitant base substitutions in the putative replicase genes of tobacco mosaic virus confer the ability to overcome the effects of a tomato resistance gene, Tm-1. EMBO J. 1988 Jun;7(6):1575–1581. doi: 10.1002/j.1460-2075.1988.tb02982.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Richards K., Guilley H., Jonard G., Hirth L. Nucleotide sequence at the 5' extremity of tobacco-mosaic-virus RNA. 1. The noncoding region (nucleotides 1-68). Eur J Biochem. 1978 Mar 15;84(2):513–519. doi: 10.1111/j.1432-1033.1978.tb12194.x. [DOI] [PubMed] [Google Scholar]
- Rietveld K., Linschooten K., Pleij C. W., Bosch L. The three-dimensional folding of the tRNA-like structure of tobacco mosaic virus RNA. A new building principle applied twice. EMBO J. 1984 Nov;3(11):2613–2619. doi: 10.1002/j.1460-2075.1984.tb02182.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Takamatsu N., Watanabe Y., Meshi T., Okada Y. Mutational analysis of the pseudoknot region in the 3' noncoding region of tobacco mosaic virus RNA. J Virol. 1990 Aug;64(8):3686–3693. doi: 10.1128/jvi.64.8.3686-3693.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tyc K., Konarska M., Gross H. J., Filipowicz W. Multiple ribosome binding to the 5'-terminal leader sequence of tobacco mosaic virus RNA. Assembly of an 80S ribosome X mRNA complex at the AUU codon. Eur J Biochem. 1984 May 2;140(3):503–511. doi: 10.1111/j.1432-1033.1984.tb08131.x. [DOI] [PubMed] [Google Scholar]
- van Belkum A., Abrahams J. P., Pleij C. W., Bosch L. Five pseudoknots are present at the 204 nucleotides long 3' noncoding region of tobacco mosaic virus RNA. Nucleic Acids Res. 1985 Nov 11;13(21):7673–7686. doi: 10.1093/nar/13.21.7673. [DOI] [PMC free article] [PubMed] [Google Scholar]