1887

Journal of General Virology header logo

Volume 69, Issue 10

Inoculation with RNAs 1 and 2 of Cucumber Mosaic Virus Induces Viral RNA Replicase Activity in Tobacco Mesophyll Protoplasts Free

Abstract

Summary

The activity of membrane-bound RNA-dependent RNA polymerase in tobacco mesophyll protoplasts was measured and products were characterized after inoculation with cucumber mosaic virus (CMV) or with different combinations of the four virus RNAs and satellite RNA. Activity increased rapidly 4 to 6 h after inoculation with virus particles. When the inoculum contained RNA 1, RNA 2 and RNA 3, most of the products of the enzyme were full-length positive-sense RNAs (RNAs 1, 2, 3 and 4) that migrated as dsRNA; inoculation of RNA 1 or RNA 2 alone did not increase RNA polymerase activity and no virus RNA was produced. After inoculation with a mixture of RNA 1 and RNA 2, however, the enzyme activity increased and full-length positive-sense RNA 1 and RNA 2 were synthesized, and we speculate that the induced enzyme is CMV RNA replicase as a complex comprising the translation products of RNA 1 and RNA 2, membrane components of the host cell and CMV RNA templates. Protoplasts inoculated with RNA 1, RNA 2 and RNA 4 did not synthesize RNA 4 which suggests that positive-sense RNA 4 does not serve as a template for the replicase complex. Satellite RNA was synthesized in protoplasts inoculated with RNA 1, RNA 2 and satellite RNA which suggests that satellite RNA is replicated by the replicase complex.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): CMV , protoplasts , replicase , RNA and satellite
© Society for General Microbiology 1988
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-69-10-2695
1988-10-01
2024-05-04
Loading full text...

Full text loading...

/deliver/fulltext/jgv/69/10/JV0690102695.html?itemId=/content/journal/jgv/10.1099/0022-1317-69-10-2695&mimeType=html&fmt=ahah

References

  1. CLARK M. F., ADAMS A. N. 1977; Characteristics of the microplate method of enzyme-linked immunosorbent assay for the detection of plant viruses. Journal of General Virology 34:475–483
     [Google Scholar]
  2. GONSALVES D., PROWIDENTI R., EDWARDS M. C. 1982; Tomato white leaf: the relation of an apparent satellite RNA and cucumber mosaic virus. Phytopathology 72:1533–1538
     [Google Scholar]
  3. GOULD A. R., SYMONS R. H. 1982; Cucumber mosaic virus RNA 3. Determination of the nucleotide sequence provides the amino acid sequences of protein 3A and viral coat protein. European Journal of Biochemistry 126:217–226
     [Google Scholar]
  4. IKEGAMI M., FRAENKEL-CONRAT H. 1979; Characterization of the RNA-dependent RNA polymerase of tobacco leaves. Journal of Biological Chemistry 254:149–154
     [Google Scholar]
  5. JASPARS E. M. J., GILL D. S., SYMONS R. H. 1985; Viral RNA synthesis by a particulate fraction from cucumber seedlings infected with cucumber mosaic virus. Virology 144:410–425
     [Google Scholar]
  6. 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]
  7. KIBERSTIS P. A., LOESCH-FRIES L. S., HALL T. C. 1981; Viral protein synthesis in barley protoplasts inoculated with native and fractionated brome mosaic virus RNA. Virology 112:804–808
     [Google Scholar]
  8. LOT H., MARCHOUX G., MARROU J., KAPER J. M., WEST C. K., VAN VLOTEN-DOTING L., HULL R. 1974; Evidence for three functional RNA species in several strains of cucumber mosaic virus. Journal of General Virology 22:81–93
     [Google Scholar]
  9. 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, U.S.A 74:4835–4838
     [Google Scholar]
  10. MANIATIS T., FRITSCH E. F., SAMBROOK J. 1982 Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Laboratory;
     [Google Scholar]
  11. MORRIS T. J., DODDS J. A. 1979; Isolation and analysis of double-stranded RNA from virus-infected plant and fungal tissue. Phytopathology 69:854–858
     [Google Scholar]
  12. MOSSOP D. W., FRANCKI R. I. B. 1979; Comparative studies on two satellite RNAs of cucumber mosaic virus. Virology 95:395–404
     [Google Scholar]
  13. NASSUTH A., BOL J. F. 1983; Altered balance of the synthesis of plus- and minus-strand RNAs induced by RNAs 1 and 2 of alfalfa mosaic virus in the absence of RNA 3. Virology 124:75–85
     [Google Scholar]
  14. NASSUTH A., ALBLAS F., BOL J. F. 1981; Localization of genetic information involved in the replication of alfalfa mosaic virus. Journal of General Virology 53:207–214
     [Google Scholar]
  15. NISHIMURA M., GRAHAM D., AKAZAWA T. 1976; Isolation of intact chloroplasts and other cell organelles from spinach leaf protoplasts. Plant Physiology 58:309–314
     [Google Scholar]
  16. OKUNO T., FURUSAWA I. 1979; RNA polymerase activity and protein synthesis in brome mosaic virus-infected protoplasts. Virology 99:218–225
     [Google Scholar]
  17. OTSUKI T., TAKEBE I. 1973; Infection of tobacco mesophyll protoplasts by cucumber mosaic virus. Virology 52:433–438
     [Google Scholar]
  18. PEDEN K. W. C., SYMONS R. H. 1973; Cucumber mosaic virus contains a functionally divided genome. Virology 53:487–492
     [Google Scholar]
  19. REZAIAN M. A., WILLIAMS R. H. V., GORDON K. H. J., GOULD A. R., SYMONS R. H. 1984; Nucleotide sequence of cucumber mosaic virus RNA 2 reveals a translation product significantly homologous to corresponding proteins of other viruses. European Journal of Biochemistry 143:277–284
     [Google Scholar]
  20. REZAIAN M. A., WILLIAMS R. H. V., SYMONS R. H. 1985; Nucleotide sequence of cucumber mosaic virus RNA 1. Presence of a sequence complementary to part of the viral satellite RNA and homologies with other viral RNAs. European Journal of Biochemistry 150:331–339
     [Google Scholar]
  21. SCHWINGHAMER M. W., SYMONS R. H. 1977; Translation of the four major RNA species of cucumber mosaic virus in plant and animal cell-free systems and in toad oocytes. Virology 79:88–108
     [Google Scholar]
  22. TAKANAMI Y. 1981; A striking change in symptoms on cucumber mosaic virus-infected tobacco plants induced by a satellite RNA. Virology 109:120–126
     [Google Scholar]
  23. TAKANAMI Y., FRAENKEL-CONRAT H. 1982a; Comparative studies on ribonucleic acid dependent RNA polymerases in cucumber mosaic virus infected cucumber and tobacco and uninfected tobacco plants. Biochemistry 21:3161–3167
     [Google Scholar]
  24. TAKANAMI Y., FRAENKEL-CONRAT H. 1982b; No viral gene is able to elicit RNA-dependent RNA polymerase in cucumber mosaic virus-infected cucumber cotyledons. Virology 116:372–374
     [Google Scholar]
  25. TAKANAMI Y., KUBO S., IMAIZUMI S. 1977; Synthesis of single- and double stranded cucumber mosaic virus RNAs in tobacco mesophyll protoplasts. Virology 80:376–389
     [Google Scholar]
  26. TAKANAMI Y., NITTA N., KUBO S. 1984; Membrane-bound RNA-dependent RNA polymerase in tobacco plants infected with cucumber mosaic virus. Abstracts, Sixth International Congress of Virolog222
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-69-10-2695
Loading
Inoculation with RNAs 1 and 2 of Cucumber Mosaic Virus Induces Viral RNA Replicase Activity in Tobacco Mesophyll Protoplasts
J. Gen. Virol. 69, 2695 (1988); https://doi.org/10.1099/0022-1317-69-10-2695
/content/journal/jgv/10.1099/0022-1317-69-10-2695
/content/journal/jgv/10.1099/0022-1317-69-10-2695
Loading

Data & Media loading...

Most cited 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