1887

Abstract

A template-bound RNA polymerase was isolated from plants infected with red clover necrotic mosaic dianthovirus (RCNMV) by differential centrifugation, solubilization with dodecyl β--maltopyranoside, and chromatography on columns of Sephacryl S-400 and Q-Sepharose. Analysis of the purified polymerase by SDS-polyacrylamide gel electrophoresis, followed by silver staining or immunoblotting, showed that it contained virus-encoded proteins of molecular masses 27 kDa and 88 kDa together with several minor proteins possibly of host origin. After removal of endogenous RNA with micrococcal nuclease, the poly-merase became template-dependent. It was also template-specific, being able to utilize as templates RNA of two strains of RCNMV, but not RNAs of three viruses in different taxonomic groups, namely cucumber mosaic cucumovirus, tomato bushy stunt tombusvirus and tomato mosaic tobamovirus. The products of RNA polymerase reactions were double-stranded RNAs corresponding to RCNMV RNAs 1 and 2. The ability of the template-dependent RNA polymerase to synthesize RNA was completely inhibited by antibodies to a peptide containing the GDD motif, whereas the activity of the template-bound enzyme was unaffected by these antibodies.

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1995-06-01
2024-04-24
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References

  1. Blumenthal T., Carmichael G. G. 1979; RNA replication: function and structure of Qβ-replicase. Annual Review of Biochemistry 48:525–548
    [Google Scholar]
  2. Brault V., Miller W. A. 1992; Translational frameshift mediated by a viral sequence in plant cells. Proceedings of the National Academy of Sciences, USA 89:2262–2266
    [Google Scholar]
  3. Buck K. W. 1986; Fungal virology: an overview. In Fungal Virology pp 1–84 Edited by Buck K. W. Boca Raton, Fla.: USA: CRC Press.;
    [Google Scholar]
  4. Candresse T., Mouches C., Bové J. M. 1986; Characterization of the virus-encoded subunit of turnip yellow mosaic virus RNA replicase. Virology 152:322–330
    [Google Scholar]
  5. Carrington J. C., Heaton L. A., Zuidema D., Hillman B. I., Morris T. J. 1989; The genome structure of turnip crinkle virus. Virology 170:219–226
    [Google Scholar]
  6. Coutts R. H., Rigden J. E., Slabas A. R., Lomonossoff G. P., Wise P. J. 1991; The complete nucleotide sequence of tobacco necrosis virus strain D. Journal of General Virology 72:1521–1529
    [Google Scholar]
  7. Dalmay T., Rubino L., Burgyan J., Kollar A., Russo M. 1993; Functional analysis of cymbidium ringspot virus genome. Virology 194:697–704
    [Google Scholar]
  8. Delarue M., Poch O., Tordo N., Moras D., Argos P. 1990; An attempt to unify the structure of polymerases. Protein Engineering 3:461–167
    [Google Scholar]
  9. Fraenkel-Conrat H. 1986; RNA-directed RNA polymerases of plants. Critical Reviews in Plant Sciences 4:213–226
    [Google Scholar]
  10. Gorbalenya A. E., Koonin E. V. 1989; Viral proteins containing the purine NTP-binding sequence pattern. Nucleic Acids Research 17:8413–8440
    [Google Scholar]
  11. Hacker D. L., Petty I. T. D., Wei N., Morris T. J. 1992; Turnip crinkle virus genes required for RNA replication and virus movement. Virology 186:1–8
    [Google Scholar]
  12. Harlow E., Lane D. 1988 Antibodies’. A Laboratory Manual New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  13. Hayes R. J., Buck K. W. 1990; Complete replication of a eukaryotic virus RNA in vitro by a purified RNA-dependent RNA polymerase. Cell 63:363–368
    [Google Scholar]
  14. Hayes R. J., Buck K. W. 1993; Analysis of replication complexes of positive strand RNA plant viruses. In Molecular Virology: A Practical Approach pp 1–34 Edited by Davison A. J., Elliott R. M. Oxford: IRL Press at Oxford University Press;
    [Google Scholar]
  15. Hayes R. J., Pereira V. C. A., McQuillin A., Buck K. W. 1994; Localization of functional regions of the cucumber mosaic virus RNA replicase using monoclonal and polyclonal antibodies. Journal of General Virology 75:3177–3184
    [Google Scholar]
  16. Hearne P. O., Knorr D. A., Hillman B. I., Morris T. J. 1990; The complete genome structure and synthesis of infectious RNA from clones of tomato bushy stunt virus. Virology 177:141–151
    [Google Scholar]
  17. Hey T. D., Richards O. C., Ehrenfeld E. 1987; Host factor-induced template modification during synthesis of poliovirus RNA in vitro. Journal of Virology 61:802–811
    [Google Scholar]
  18. Houwing C. J., Jaspars E. M. J. 1986; Coat protein blocks the in vitro transcription of virion RNAs of alfalfa mosaic virus. FEBS Letters 209:284–288
    [Google Scholar]
  19. Jacoba-Molina A., Ding J., Nanni R. G., Clark A. D., Lu X., Tantillo C., Williams R. L., Kamer G., Ferris A. L., Clark P., Hizi A., Hughes S. H., Arnold E. 1993; Crystal structure of human immunodeficiency virus type 1 reverse transcriptase complexed with double-stranded DNA at 3.0 Å resolution shows bent DNA. Proceedings of the National Academy of Sciences, USA 90:6320–6324
    [Google Scholar]
  20. Kamer G., Argos P. 1984; Primary structural comparison of RNA dependent polymerases from plant, animal and bacterial viruses. Nucleic Acids Research 12:7269–7282
    [Google Scholar]
  21. Kim K. H., Lommel S. A. 1994; Identification and analysis of the site of 1 ribosomal frameshifting in red clover necrotic mosaic virus. Virology 200:574–582
    [Google Scholar]
  22. Kohstaedt L. A., Wang J., Friedman J. M., Rice P. A., Steitz T. A. 1992; Crystal structure at 3.5 Å resolution of HIV-1 reverse transcriptase complexed with an inhibitor. Science 256:1783–1790
    [Google Scholar]
  23. Kollar A., Burgyan J. 1994; Evidence that ORF 1 and 2 are the only virus-encoded replicase genes of cymbidium ringspot tombus-virus. Virology 201:169–172
    [Google Scholar]
  24. Koonin E. V. 1991; The phylogeny of RNA-dependent RNA polymerases of positive-strand RNA viruses. Journal of General Virology 72:2197–2206
    [Google Scholar]
  25. Koonin E. V., Dolja V. V. 1993; Evolution and taxonomy of positive-strand RNA viruses: implications of comparative analysis of amino acid sequences. Critical Reviews in Biochemistry and Molecular Biology 28:375–430
    [Google Scholar]
  26. Kozak M. 1992; A consideration of alternative models for the initiation of translation in eukaryotes. Critical Reviews in Biochemistry and Molecular Biology 27:385–102
    [Google Scholar]
  27. Laemmli U. K. 1970; Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  28. Lommel S. A., Weston-Fina M., Xiong Z., Lomonossoff G. P. 1988; The nucleotide sequence and gene organization of red clover necrotic mosaic virus RNA-2. Nucleic Acids Research 16:8587–8602
    [Google Scholar]
  29. Maina C. V., Riggs P. D., Grandea A. G., Slatko B. E., Moran L. S., Tagliamonte J. A., McReynolds L. A., Guan C. 1988; A vector to express and purify foreign proteins in Escherichia coli by fusion to, and separation from, maltose binding protein. Gene 74:365–373
    [Google Scholar]
  30. Merrifield R. B. 1963; Solid phase peptide synthesis. I. The synthesis of a tetrapeptide. Science 85:2149–2154
    [Google Scholar]
  31. Miller W. A., Hall T. C. 1983; Use of micrococcal nuclease in the purification of highly template dependent RNA polymerase from brome mosaic virus-infected barley. Virology 125:236–241
    [Google Scholar]
  32. Musil M. 1969; Serological properties of certain isolates of red clover necrotic mosaic virus. Acta Virologica 13:226–234
    [Google Scholar]
  33. Ollis D. L., Brick P., Hamlin R., Xuong N. G., Steitz T. A. 1985; Structure of the large fragment of Escherichia coli DNA polymerase I complexed with dTMP. Nature 313:762–766
    [Google Scholar]
  34. Osman T. A. M., Buck K. W. 1987; Replication of red clover necrotic mosaic virus RNA in cowpea protoplasts: RNA 1 replicates independently of RNA 2. Journal of General Virology 68:289–296
    [Google Scholar]
  35. Osman T. A. M., Buck K. W. 1991; Detection of the movement protein of red clover necrotic mosaic virus in a cell wall fraction from infected Nicotiana clevelandii plants. Journal of General Virology 72:2853–2856
    [Google Scholar]
  36. Osman T. A. M., Dodd S. M., Buck K. W. 1986; RNA 2 of red clover necrotic mosaic virus determines lesion morphology and systemic invasion in cowpea. Journal of General Virology 67:203–207
    [Google Scholar]
  37. Paje-Manalo L. L., Lommel S. A. 1989; Independent replication of red clover necrotic mosaic virus electroporated host and non-host Nicotiana sp. protoplasts. Phytopathology 79:457–461
    [Google Scholar]
  38. Quadt R., Jaspars E. M. J. 1990; Purification and characterization of brome mosaic virus RNA-dependent RNA polymerase. Virology 178:189–194
    [Google Scholar]
  39. Quadt R., Jaspars E. M. J. 1991; Characterization of cucumber mosaic virus RNA-dependent RNA polymerase. FEBS letters 279:273–276
    [Google Scholar]
  40. Quadt R., Rosdorff H. J. M., Hunt T. W., Jaspars E. M. J. 1991; Analysis of the protein composition of alfalfa mosaic virus RNA-dependent RNA polymerase. Virology 182:309–315
    [Google Scholar]
  41. Quadt R., Kao C. C., Browning K. S., Hershberger R. P., Ahlquist P. 1993; Characterization of a host protein associated with brome mosaic virus RNA-dependent RNA polymerase. Proceedings of the National Academy of Sciences, USA 90:1498–1502
    [Google Scholar]
  42. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: A Laboratory Manual 2nd edn New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  43. Saunders K., Kaesberg P. 1985; Template-dependent RNA polymerase from black beetle virus-infected Drosophila melanogaster cells. Virology 144:373–381
    [Google Scholar]
  44. Schiebel W., Haas B., Marinkovic S., Klanner A., Sänger H. L. 1993; RNA-directed RNA polymerase from tomato leaves. I. Purification and physical properties. Journal of Biological Chemistry 263:11851–11857
    [Google Scholar]
  45. Scholthof H. B., Morris T. J., Jackson A. O. 1993; The capsid protein gene of tomato bushy stunt virus is dispensable for systemic movement and can be replaced for localized expression of foreign genes. Molecular Plant-Microbe Interactions 6:309–322
    [Google Scholar]
  46. Song C., Simon A. E. 1994; RNA-dependent RNA polymerase from plants infected with turnip crinkle virus can transcribe (+)- and (−)-strands of virus-associated RNAs. Proceedings of the National Academy of Sciences, USA 91:8792–8796
    [Google Scholar]
  47. Sousa R., Chung Y. J., Rose J. P., Wang B. C. 1993; Crystal structure of bacteriophage T7 RNA polymerase at 3-3 A resolution. Nature 364:593–599
    [Google Scholar]
  48. Wu S., Kaesberg P. 1991; Synthesis of template-sense, singlestrand flockhouse virus RNA in a cell-free replication system. Virology 183:392–396
    [Google Scholar]
  49. Xiong Z., Lommel S. A. 1989; The complete nucleotide sequence and genome organization of red clover necrotic mosaic virus RNA-1. Virology 171:543–554
    [Google Scholar]
  50. Xiong Z., Kim K. H., Kendall T. L., Lommel S. A. 1993; Synthesis of the putative red clover necrotic mosaic virus RNA polymerase by ribosomal frameshifting in vitro. Virology 193:213–221
    [Google Scholar]
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