1887

Abstract

SUMMARY

The RNA molecules of 13 plant viruses were denatured by heating at 50 °C in 1 -glyoxal, 50% dimethyl sulphoxide, and their mol. wt. (expressed as the sodium salts) estimated both by electrophoresis in 0.75% agarose gel, and by measuring their lengths in the electron microscope. The two methods gave similar results. Compared with previous estimates obtained by electrophoresis at 60 °C in 2% polyacrylamide gels containing 8 -urea, the new estimates were similar for molecules of mol. wt. up to about 1.7 × 10 (about 5000 nucleotides) but became increasingly larger as the mol. wt. exceeded this value. The mol. wt. of the RNA-1 molecules of a range of nepoviruses were all found to be about 2.7 × 10 to 2.8 × 10 (about 7800 to 8100 nucleotides) by the glyoxal/agarose method compared with about 2.1 × 10 to 2.2 × 10 (about 6100 to 6400 nucleotides) by the hot urea/polyacrylamide method. The estimated mol. wt. of RNA-1 (B-RNA) of cowpea mosaic virus (CPMV) was 2.39 × 10 (6900 nucleotides), about 20% greater than the commonly used value, those of the single RNA species of apple chlorotic leafspot and heracleum latent viruses were 2.48 × 10 (7170 nucleotides) and 2.52 × 10 (7260 nucleotides) respectively and that of parsnip yellow fleck virus was 3.34 × 10 (9650 nucleotides). Values of 2.37 × 10 (6840 nucleotides) and 0.54 × 10 (1550 nucleotides) were found respectively for RNA-1 and RNA-2 of the CAM strain of tobacco rattle virus (TRV). For the nepoviruses, and for CPMV and TRV, the new mol. wt. estimates were in good agreement with those predicted from the morphology or sedimentation behaviour of the virus particles. Electrophoresis of glyoxylated RNA in agarose gels is preferred to other electrophoretic methods for determining the mol. wt. of RNA molecules larger than about 1.7 × 10 (about 5000 nucleotides), and appears reliable for mol. wt. up to at least about 3.5 × 10 (10000 nucleotides).

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1981-04-01
2022-01-25
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References

  1. Allen W. R., Dias H. F. 1977; Properties of the single protein and two nucleic acids of tomato ringspot virus. Canadian Journal of Botany 55:1028–1037
    [Google Scholar]
  2. Bailey J. M., Davidson N. 1976; Methylmercury as a reversible denaturing agent for agarose gel electrophoresis. Analytical Biochemistry 70:75–85
    [Google Scholar]
  3. Bem F., Murant A. F. 1979; Host range, purification and serological properties of heracleum latent virus. Annals of Applied Biology 92:243–256
    [Google Scholar]
  4. Boedtker H. 1960; Configurational properties of tobacco mosaic virus ribonucleic acid. Journal of Molecular Biology 2:171–188
    [Google Scholar]
  5. Boedtker H. 1971; Conformation independent molecular weight determinations of RNA by gel electrophoresis. Biochimica et Biophysica Acta 240:448–453
    [Google Scholar]
  6. Boedtker H., Crkvenjakov R. B., Dewey K. F., Lanks K. 1973; Some properties of high molecular weight ribonucleic acid isolated from chick embryo polysomes. Biochemistry 12:4356–4360
    [Google Scholar]
  7. Brosius J., Palmer M. L., Kennedy P. J., Noller H. F. 1978; Complete nucleotide sequence of a 16S ribosomal RNA gene from Escherichia coli . Proceedings of the National Academy of Sciences of the United States of America 75:4801–4805
    [Google Scholar]
  8. Brosius J., Dull T. J., Noller H. F. 1980; Complete nucleotide sequence of a 23 S ribosomal RNA gene from Escherichia coli . Proceedings of the National Academy of Sciences of the United States of America 77:201–204
    [Google Scholar]
  9. Carbon P., Ehresmann C., Ehresmann B., Ebel P. 1978; The sequence of Escherichia coli 16 S RNA determined by new rapid gel methods. FEBS Letters 94:152–156
    [Google Scholar]
  10. Caspar D. L. D. 1963; Assembly and stability of the tobacco mosaic virus particle. Advances in Protein Chemistry 18:37–121
    [Google Scholar]
  11. Cohn E. J., Edsall J. T. (editors) 1943 Proteins, Amino Acids and Peptides as Ions and Dipolar Ions pp 370–381 New York: Reinhold;
    [Google Scholar]
  12. Cooper J. I., Mayo M. A. 1972; Some properties of the particles of three tobravirus isolates. Journal of General Virology 16:285–297
    [Google Scholar]
  13. Davis R. W., Simon M., Davidson N. 1971; Electron microscope heteroduplex methods for mapping regions of base sequence homology in nucleic acids. Methods in Enzymology 21D:413–428
    [Google Scholar]
  14. Dunez J., Delbos R., Dupont G. 1976; Myrobalan latent ringspot virus. Commonwealth Mycological Institute/Association of Applied Biologists Descriptions of Plant Viruses No. 160
    [Google Scholar]
  15. Geelen J. L. M. C., van Kammen A., Verduin B. J. M. 1972; Structure of the capsid of cowpea mosaic virus. The chemical subunit: molecular weight and number of subunits per particle. Virology 49:205–213
    [Google Scholar]
  16. Gooding G. V., Hebert T. T. 1967; A simple technique for purification of tobacco mosaic virus in large quantities. Phytopathology 57:1285
    [Google Scholar]
  17. Grivell L. A., Reijnders L., Borst P. 1971; The effect of temperature and ionic strength on the electrophoretic mobility of yeast mitochondrial RNA. European Journal of Biochemistry 19:64–72
    [Google Scholar]
  18. Harrison B. D., Woods R. D. 1966; Serotypes and particle dimensions of tobacco rattle viruses from Europe and North America. Virology 28:610–620
    [Google Scholar]
  19. Hsu M. T., Kung H. J., Davidson N. 1973; An electron microscope study of Sindbis virus RNA. Cold Spring Harbor Symposia on Quantitative Biology 38:943–950
    [Google Scholar]
  20. Tones A. T., Mayo M. A. 1972; The two nucleoprotein particles of cherry leaf roll virus. Journal of General Virology 16:349–358
    [Google Scholar]
  21. Lehrach H., Diamond D., Wozney J. M., Boedtker H. 1977; RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical re-examination. Biochemistry 16:4743–4751
    [Google Scholar]
  22. Lister R. M., Hadidi A. F. 1971; Some properties of apple chlorotic leaf spot virus and their relation to purification problems. Virology 45:240–251
    [Google Scholar]
  23. Lister R. M., Bancroft J. B., Nadakavukaren M. J. 1965; Some sap-transmissible viruses from apple. Phytopathology 55:859–870
    [Google Scholar]
  24. Mcmaster G. K., Carmichael G. G. 1977; Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proceedings of the National Academy of Sciences of the United States of America 74:4835–4838
    [Google Scholar]
  25. Markham R. 1962; The analytical ultracentrifuge as a tool for the investigation of plant viruses. Advances in Virus Research 9:241–270
    [Google Scholar]
  26. Mayo M. A., Murant A. F., Harrison B. D. 1971; New evidence on the structure of nepoviruses. Journal of General Virology 12:175–178
    [Google Scholar]
  27. Mayo M. A., Murant A. F., Harrison B. D., Goold R. A. 1974; Two protein and two RNA species in particles of strawberry latent ringspot virus. Journal of General Virology 24:29–37
    [Google Scholar]
  28. Murant A. F. 1970a; Arabis mosaic virus. CMI/AAB Descriptions of Plant Viruses No. 16
    [Google Scholar]
  29. Murant A. F. 1970b; Tomato black ring virus. CMI/AAB Descriptions of Plant Viruses No. 38
    [Google Scholar]
  30. Murant A. F., Goold R. A. 1968; Purification, properties and transmission of parsnip yellow fleck, a semi-persistent, aphid-borne virus. Annals of Applied Biology 62:123–137
    [Google Scholar]
  31. Murant A. F., Taylor M. 1978; Estimates of molecular weights of nepovirus RNA species by polyacrylamide gel electrophoresis under denaturing conditions. Journal of General Virology 41:53–61
    [Google Scholar]
  32. Murant A. F., Mayo M. A., Harrison B. D., Goold R. A. 1972; Properties of virus and RNA components of raspberry ringspot virus. Journal of General Virology 16:327–338
    [Google Scholar]
  33. Murant A. F., Mayo M. A., Harrison B. D., Goold R. A. 1973; Evidence for two functional RNA species and a ‘satellite’ RNA in tomato black ring virus. Journal of General Virology 19:275–278
    [Google Scholar]
  34. Murant A. F., Taylor M., Duncan G. H. 1979; Estimates of molecular weight of virus RNA by electrophoresis and electron microscopy. Annual Report of the Scottish Horticultural Research Institute for 1978: pp 98–99
    [Google Scholar]
  35. Pinder J. C., Staynov D. Z., Gratzer W. B. 1974; Electrophoresis of RNA in formamide. Biochemistry 13:5373–5378
    [Google Scholar]
  36. Reijnders L., Sloof P., Sival J., Borst P. 1973; Gel electrophoresis of RNA under denaturing conditions. Biochimica et Biophysica Acta 324:320–333
    [Google Scholar]
  37. Reijnders L., Aalbers A. M. J., Van Kammen A., Touring R. W. J. 1974; Molecular weights of plant virus RNAs determined by gel electrophoresis under denaturing conditions. Virology 60:515–521
    [Google Scholar]
  38. Ross G. J. S. 1975; Simple non-linear modelling for the general user. Bulletin of the International Statistical Institute 46:585–593
    [Google Scholar]
  39. Schneider I. R., Diener T. O. 1966; The correlation between the proportions of virus-related products and the infectious component during the synthesis of tobacco ringspot virus. Virology 29:92–99
    [Google Scholar]
  40. Schneider I. R., White R. M., Civerolo E. L. 1974; Two nucleic acid-containing components of tomato ringspot virus. Virology 57:139–146
    [Google Scholar]
  41. Spohr G., Mirault M. E., Imaizumi T., Scherrer K. 1976; Molecular weight determination of animal cell RNA by electrophoresis in formamide under fully denaturing conditions on exponential polyacrylamide gels. European Journal of Biochemistry 62:313–322
    [Google Scholar]
  42. Stace-Smith R. 1966; Purification and properties of tomato ringspot virus and an RNA-deficient component. Virology 29:240–247
    [Google Scholar]
  43. Stace-Smith R., Reichmann M. E., Wright N. S. 1965; Purification and properties of tobacco ringspot virus and two RNA-deficient components. Virology 25:487–494
    [Google Scholar]
  44. Tremaine J. H., Stace-Smith R. 1968; Chemical composition and biophysical properties of tomato ringspot virus. Virology 35:102–107
    [Google Scholar]
  45. Van Kammen A. 1967; Purification and properties of the components of cowpea mosaic virus. Virology 31:633–642
    [Google Scholar]
  46. Walkey D. G. A., Stace-Smith R., Tremaine J. H. 1973; Serological, physical and chemical properties of strains of cherry leaf roll virus. Phytopathology 63:566–571
    [Google Scholar]
  47. Wellauer P. K., Dawid I. B. 1973; Secondary structure maps of RNA: processing of HeLa ribosomal RNA. Proceedings of the National Academy of Sciences of the United States of America 70:2827–2831
    [Google Scholar]
  48. Wu G. J., Bruening G. 1971; Two proteins from cowpea mosaic virus. Virology 46:596–612
    [Google Scholar]
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