1887

Abstract

Destabilizing events required for subsequent cotranslational disassembly of tobacco mosaic virus (TMV) particles were studied. Brief treatment of U-P-labelled TMV (strain or U2) with 1% SDS exposed only 2.5% of the RNA (160 5′ nucleotides) in a susceptible subpopulation of virions. Limited uncoating occurred almost immediately and appeared to be synchronous because the amount of 5′ oligonucleotide marker () recovered remained constant throughout a 15 min period in SDS. Additional RNase T-sensitive oligonucleotides were exposed only after 1 to 2 min in SDS. Coat protein (CP) subunits released from virions ‘destabilized’ by ultracentrifugation at between pH 7.2 and 9.2 were quantified using -[S]methionine-labelled particles of TMV strain U2. CP recovery and virus particle translation results were consistent with increasing numbers of virions uncoating for approximately 200 nucleotides. In the presence of sparsomycin (SPN), the TMV strain 5′ leader and the first AUG codon can bind two 80S ribosomes. Electron microscopy of pH 7.5-treated TMV particles incubated in SPN-treated wheatgerm extract or rabbit reticulocyte lysate, showed that approximately 10% of virions complexed with one ribosome and approximately 10% with two bound ribosomes, confirming that at least had been uncoated. Nucleocapsids in these complexes were shorter than untreated TMV by 9 to 10 nm (i.e. equivalent to 192 to 217 nucleotides exposed). The template activities of virions pretreated at pH 7.2 to 9.2 were destroyed by RNase H when short cDNAs were hybridized to sequences at, or immediately 3′ to, the first AUG codon. We propose that the complete 5′ leader of TMV RNA interacts weakly with CP subunits and that this micro-instability is due to the absence of G residues and is essential for initiation of cotranslational virus disassembly.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-72-4-769
1991-04-01
2022-12-04
Loading full text...

Full text loading...

/deliver/fulltext/jgv/72/4/JV0720040769.html?itemId=/content/journal/jgv/10.1099/0022-1317-72-4-769&mimeType=html&fmt=ahah

References

  1. AbouHaidar M. G., Erickson J. W. 1985; Structure and in vitro assembly of papaya mosaic virus. In Molecular Plant Virology vol 1 pp 85–121 Edited by Davies J. W. Boca Raton: CRC Press;
    [Google Scholar]
  2. Atabekov K. J., Tyulkina L. G., Karpova O. V., Metelev V. G., Rodionova N. P., Shabarova Z. A., Atabekov J. G. 1988; Site-specific enzymatic cleavage of TMV RNA directed by deoxyribo- and chimeric (deoxyribo-ribo) oligonucleotides. FEBS Letters 232:96–98
    [Google Scholar]
  3. Blowers L. E., Wilson T. M. A. 1982; The effect of urea on tobacco mosaic virus: polarity of disassembly. Journal of General Virology 61:137–141
    [Google Scholar]
  4. Brisco M. J., Hull R., Wilson T. M. A. 1985; Southern bean mosaic virus-specific proteins are synthesized in an in vitro system supplemented with intact, treated virions. Virology 143:392–398
    [Google Scholar]
  5. Brisco M. J., Hull R., Wilson T. M. A. 1986; Swelling of isometric and of bacilliform plant virus nucleocapsids is required for virus-specific protein synthesis in vitro. Virology 148:210–217
    [Google Scholar]
  6. Bruening G., Beachy R. N., Scalla R., Zaitlin M. 1976; In vitro and in vivo translation of the ribonucleic acids of a cowpea strain of tobacco mosaic virus. Virology 71:498–517
    [Google Scholar]
  7. Crum C., Johnson J. D., Nelson A., Roth D. 1988; Complementary oligodeoxynucleotide mediated inhibition of tobacco mosaic virus RNA translation in vitro. Nucleic Acids Research 16:4569–4581
    [Google Scholar]
  8. Davies J. W. 1979; Translation of plant virus ribonucleic acids in extracts from eukaryotic cells. In Nucleic Acids in Plants vol II pp 111–149 Edited by Hall T. C., Davies J. W. Boca Raton: CRC Press;
    [Google Scholar]
  9. Filipowicz W., Haenni A.-L. 1979; Binding of ribosomes to 5″-terminal leader sequences of eukaryotic messenger RNAs. Proceedings of the National Academy of Sciences U.S.A. 76:3111–3115
    [Google Scholar]
  10. Gallie D. R., Sleat D. E., Watts J. W., Turner P. C., Wilson T. M. A. 1987; In vivo uncoating and efficient expression of foreign mRNAs packaged in TMV-like particles. Science 236:1122–1124
    [Google Scholar]
  11. Garfin D. E., Mandeles S. 1975; Sequences of oligonucleotides prepared from tobacco mosaic virus ribonucleic acid. Virology 64:388–399
    [Google Scholar]
  12. Goelet P., Lomonossoff G. P., Butler P. J. G., Akam M. E., Gait M. J., Karn J. 1982; Nucleotide sequence of tobacco mosaic virus RNA. Proceedings of the National Academy of Sciences, U.S.A 79:5818–5822
    [Google Scholar]
  13. Hogue R., Asselin A. 1984; Study of tobacco mosaic virus in vitro disassembly by sucrose density gradient centrifugation and agarose gel electrophoresis. Canadian Journal of Botany 62:457–462
    [Google Scholar]
  14. Jupin I., Sleat D. E., Watkins P. A. C., Wilson T. M. A. 1989; Direct recovery of in vitro transcripts in a protected form suitable for prolonged storage and shipment at ambient temperatures.. Nucleic Acids Research 17:815
    [Google Scholar]
  15. Kozak M. 1989; The scanning model for translation: an update. Journal of Cell Biology 108:229–241
    [Google Scholar]
  16. Kukla B. A., Guilley H. A., Jonard G. X., Richards K. E., Mundry K. W. 1979; Characterization of long guanosine-free RNA sequences from the Dahlemense and U2 strains of tobacco mosaic virus. European Journal of Biochemistry 98:61–66
    [Google Scholar]
  17. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  18. Leberman R. 1966; The isolation of plant viruses by means of ‘simple’ coacervates. Virology 30:341–347
    [Google Scholar]
  19. Mandeles S. 1968; Location of unique sequences in tobacco mosaic virus ribonucleic acid. Journal of Biological Chemistry 243:3671–3674
    [Google Scholar]
  20. Minshull J., Hunt T. 1986; The use of single-stranded DNA and RNase H to promote quantitative ‘hybrid arrest of translation’ of mRNA/DNA hybrids in reticulocyte lysate cell-free translations. Nucleic Acids Research 14:6433–6451
    [Google Scholar]
  21. Mundry K. W. 1965; A model of the coat protein cistron of tobacco mosaic virus and its biochemical investigation: the model, the experimental approach, and the isolation of a long oligonucleotide from TMV-RNA. Zeitschrift für Vererbungslehre 97:281–296
    [Google Scholar]
  22. Mundry K. W. 1967; Strain-specific sequences in TMV-RNA. Molecular and General Genetics 100:383–384
    [Google Scholar]
  23. Mundry K. W. 1969; Structural elements of viral ribonucleic acid and their variation. I. An adenine-rich and strain-specific segment in tobacco mosaic virus ribonucleic acid. Molecular and General Genetics 105:361–377
    [Google Scholar]
  24. Mundry K. W., Priess H. 1971; Structural elements of viral ribonucleic acid and their variation. II. 32P-oligonucleotide maps of large G-lacking segments of RNA of tobacco mosaic virus wild strains. Virology 46:86–97
    [Google Scholar]
  25. Namba K., Stubbs G. 1986; Structure of tobacco mosaic virus at 3.6Å resolution: implications for assembly. Science 231:1401–1406
    [Google Scholar]
  26. Namba K., Pattanayek R., Stubbs G. 1989; Visualization of protein-nucleic acid interactions in a virus. Refined structure of intact tobacco mosaic virus at 2.9Å resolution by X-ray fibre diffraction. Journal of Molecular Biology 29:431–437
    [Google Scholar]
  27. Pelcher L. E., Halasa M. C. 1979a; Factors influencing the production of intermediate particles during alkaline degradation of tobacco mosaic virus: time, pH, salt concentration, and temperature. Journal of Virology 29:431–437
    [Google Scholar]
  28. Pelcher L. E., Halasa M. C. 1979b; A comparative study of the alkaline disassembly of two strains of tobacco mosaic virus. Virology 98:489–492
    [Google Scholar]
  29. Pelcher L. E., Walmsley S. L., Mackenzie S. L. 1980; The effects of heterologous and homologous coat protein on alkaline disassembly of tobacco and tomato isolates of tobacco mosaic virus. Virology 105:287–290
    [Google Scholar]
  30. Pelham H. R. B., Jackson R. J. 1976; An efficient mRNA-dependent translation system from reticulocyte lysates. European Journal of Biochemistry 67:247–256
    [Google Scholar]
  31. Perham R. N. 1969; Sucrose density-gradient analysis of the alkaline degradation of tobacco mosaic virus. Journal of Molecular Biology 45:439–441
    [Google Scholar]
  32. Perham R. N., Wilson T. M. A. 1976; The polarity of stripping of coat protein subunits from the RNA in tobacco mosaic virus under alkaline conditions. FEBS Letters 62:11–15
    [Google Scholar]
  33. Perham R. N., Wilson T. M. A. 1978; The characterization of intermediates formed during the disassembly of tobacco mosaic virus at alkaline pH. Virology 84:293–302
    [Google Scholar]
  34. Richards K., Guilley H., Jonard G., Hirth L. 1978; Nucleotide sequence at the 5′ extremity of tobacco-mosaic-virus RNA. 1. The noncoding region (nucleotides 1-68). European Journal of Biochemistry 84:513–519
    [Google Scholar]
  35. Roenhorst J. W., Verduin B. J. M., Goldbach R. W. 1989; Virus-ribosome complexes from cell-free translation systems supplemented with cowpea chlorotic mottle virus particles. Virology 168:138–146
    [Google Scholar]
  36. Schuster H., Wilhelm R. C. 1963; Reaction differences between TMV and its free nucleic acid with nitrous acid. Biochimica et biophysica acta 68:554–560
    [Google Scholar]
  37. Sehgal O. P., Soong M. M. 1972; Reaction of nitrous acid with viral nucleic acids in situ.. Virology 47:239–243
    [Google Scholar]
  38. Shaw J. G., Plaskitt K. A., Wilson T. M. A. 1986; Evidence that tobacco mosaic virus particles disassemble cotranslationally in vivo. Virology 148:326–336
    [Google Scholar]
  39. Shields S. A., Brisco M. J., Wilson T. M. A., Hull R. 1989; Southern bean mosaic virus RNA remains associated with swollen virions during translation in wheat germ cell-free extracts. Virology 171:602–606
    [Google Scholar]
  40. Shirako Y., Ehara Y. 1986; Comparison of the in vitro translation products of wild-type and a deletion mutant of soil-borne wheat mosaic virus. Journal of General Virology 67:1237–1245
    [Google Scholar]
  41. Sleat D. E., Turner P. C., Finch J. T., Butler P. J. G., Wilson T. M. A. 1986; Packaging of recombinant RNA molecules into pseudovirus particles directed by the origin-of-assembly sequence from tobacco mosaic virus RNA. Virology 155:299–308
    [Google Scholar]
  42. Sleat D. E., Hull R., Turner P. C., Wilson T. M. A. 1988a; Studies on the mechanism of translational enhancement by the 5′-leader sequence of tobacco mosaic virus RNA. European Journal of Biochemistry 175:75–86
    [Google Scholar]
  43. Sleat D. E., Gallie D. R., Watts J. W., Deom C. M., Turner P. C., Beachy R. N., Wilson T. M. A. 1988b; Selective recovery of foreign gene transcripts as virus-like particles in TMV-infected transgenic tobaccos. Nucleic Acids Research 16:3127–3140
    [Google Scholar]
  44. Solís I., García-Arenal F. 1990; The complete nucleotide sequence of the genomic RNA of the tobamovirus tobacco mild green mosaic virus. Virology 177:553–558
    [Google Scholar]
  45. Tyc K., Konarska M., Gross H. J., Filipowicz W. 1984; Multiple ribosome binding to the 5′-terminal leader sequence of tobacco mosaic virus RNA. Assembly of an 80S ribosome mRNA complex at the AUU codon. European Journal of Biochemistry 140:503–511
    [Google Scholar]
  46. Wilson T. M. A. 1984a; Cotranslational disassembly of tobacco mosaic virus in vitro. Virology 137:255–265
    [Google Scholar]
  47. Wilson T. M. A. 1984b; Cotranslational disassembly increases the efficiency of expression of TMV RNA in wheat germ cell-free extracts. Virology 138:353–356
    [Google Scholar]
  48. Wilson T. M. A. 1988; Structural interactions between plant RNA viruses and cells. In Oxford Surveys of Plant Molecular and Cell Biology vol 5 pp 89–144 Edited by Miflin B. J. Oxford: Oxford University Press;
    [Google Scholar]
  49. Wilson T. M. A., Shaw J. G. 1987; Cotranslational disassembly of filamentous plant virus nucleocapsids in vitro and in vivo.. In Positive Strand RNA Viruses, UCLA Symposia on Molecular and Cellular Biology, New Series vol 54 pp 159–181 Edited by Brinton M. A., Rueckert R. R. New York: Alan R. Liss;
    [Google Scholar]
  50. Wilson T. M. A., Perham R. N., Finch J. T., Butler P. J. G. 1976; Polarity of the RNA in the tobacco mosaic virus particle and the direction of protein stripping in sodium dodecyl sulphate. FEBS Letters 64:285–289
    [Google Scholar]
  51. Wilson T. M. A., Lomonossoff G. P., Glover J. F. 1981; Dimethyl sulphoxide (DMSO) disassembles tobacco mosaic virus predominantly from the 5′-end of the viral RNA. Journal of General Virology 53:225–234
    [Google Scholar]
  52. Wilson T. M. A., Plaskitt K. A., Watts J. W., Osbourn J. K., Watkins P. A. C. 1990; Signals and structures involved in early interactions between plants and viruses or pseudoviruses. In Recognition and Response in Plant-Virus Interactions, NATO ASI Series vol H41 pp 123–145 Edited by Fraser R. S. S. Berlin & Heidelberg: Springer-Verlag;
    [Google Scholar]
  53. Wittmann H. G. 1965; Die primare Proteinstruktur von Stammen des Tabakmosaikvirus. Teil IV. Aminosauresequenzen (Pos. 1–61 und 135–158) des Proteins des Tabakmosaikvirusstammes U2. Zeitschrift für Naturforschung 20b:1213–1223
    [Google Scholar]
  54. Zimmern D., Wilson T. M. A. 1976; Location of the origin for viral reassembly on tobacco mosaic virus RNA and its relation to stable fragment. FEBS Letters 71:294–298
    [Google Scholar]
  55. Zuker M., Stiegler P. 1981; Optimal computer folding of large RNA sequences using thermodynamics and auxiliary information. Nucleic Acids Research 9:133–148
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-72-4-769
Loading
/content/journal/jgv/10.1099/0022-1317-72-4-769
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