1887

Abstract

The 16.7 kbp dsRNA specific to the ‘447’ cytoplasmic male sterility (CMS) line of was labelled with [α-P]ATP and poly(A) polymerase, and by T4 RNA ligase-mediated addition of [P]pCp. Analysis of the reaction products under denaturing conditions revealed in both cases extensive labelling of a 4.5 kb ssRNA, already detected in previous experiments in which the RNA-dependent RNA polymerase associated with the dsRNA was allowed to pursue RNA synthesis on preinitiated complexes. Mobility shift analysis of total pCp-labelled dsRNA revealed not two but three different 3′ termini. The most prominent sequencing pattern corresponded to the 4.5 kb ssRNA, indicating that this RNA species has a preferentially accessible, free 3′ OH extremity. Northern blot analysis of the denatured dsRNA confirmed that the 4.5 kb ssRNA is a subgenomic mRNA and detected its counterpart of about 12 kb. Nearly all 16.7 kbp dsRNA molecules featured an interrupted positive-sense strand, indicating a marked prevalence of transcription over replication complexes. This unusual strategy of transcription by a strand displacement mechanism, following initiation at an internal discontinuity, is compared with that of other dsRNA viruses or defective viruses, and is discussed in relation to the expression of the CMS trait.

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1993-06-01
2021-10-26
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References

  1. Belliard G., Vedel F., Pelletier G. 1979; Mitochondrial recombination in cytoplasmic hybrids of Nicotiana tabacum by protoplast fusion. Nature, London 281:401–403
    [Google Scholar]
  2. Boccardo G., Milne R. G., Luisoni E., Lisa V., Accotto G. P. 1985; Three seedborne cryptic viruses containing double-stranded RNA isolated from white clover. Virology 147:29–40
    [Google Scholar]
  3. Boeshore M. L., Hanson M. L., Izhar S. 1985; A variant mitochondrial DNA arrangement specific to petunia stable sterile somatic hybrid. Plant Molecular Biology 4:125–132
    [Google Scholar]
  4. Choi G. H., Nuss D. L. 1992; Hypovirulence of chestnut blight fungus conferred by an infectious viral cDNA. Science 257:800–803
    [Google Scholar]
  5. Dewey R. E., III Levings C. S., Timothy D. H. 1986; Novel recombinations in the maize mitochondrial genome produce a unique transcriptional unit in the Texas male-sterile cytoplasm. Cell 44:439–449
    [Google Scholar]
  6. Donis-Keller H., Maxam A. M., Gilbert W. 1977; Mapping adenines, guanines and pyrimidines in RNA. Nucleic Acids Research 4:2527–2538
    [Google Scholar]
  7. Duc G., Scalla R., Lefebvre A. 1984; New developments in cytoplasmic male sterility in Vicia faba. In Vida faba Agronomy, Physiology and Breeding pp. 254–260 Edited by Hebble-thwaite P. D., Dawkins T. C. K., Heath M. C., Lockwood G. The Hague: Martinus Nijhoff;
    [Google Scholar]
  8. Dulieu P., Penin P., Dulieu H., Gautheron D. C. 1988; Purification of virus-like particles from Vicia faba and detection by ELISA in crude leaf extracts. Plant Science 56:9–14
    [Google Scholar]
  9. Edwardson J. R., Bond D. A., Christie R. G. 1976; Cytoplasmic sterility factors in Vicia faba L. Genetics 82:443–449
    [Google Scholar]
  10. England T., Uhlenbeck O. 1978; 3′-terminal labelling of RNA with T4 RNA ligase. Nature, London 275:560–561
    [Google Scholar]
  11. Finnegan P. M., Brown G. G. 1986; Autonomously replicating RNA in mitochondria of maize plants with S-type cytoplasm. Proceedings of the National Academy of Sciences, U.S.A. 83:5175–5179
    [Google Scholar]
  12. Gargouri R., Joshi R. L., Bol J. F., Astier-Manifacier S., Haenni A.-L. 1989; Mechanism of synthesis of turnip yellow mosaic virus protein subgenomic RNA in vivo. Virology 171:386–393
    [Google Scholar]
  13. Grill L. K., Garger S. J. 1981; Identification and characterization of double stranded RNA associated with cytoplasmic male sterility in Vicia faba. Proceedings of the National Academy of Sciences, U.S.A. 78:7043–7046
    [Google Scholar]
  14. Gustavson G. D., Milner J. J., McFarland J. E., Pedersen K., Larkins B. A., Jackson A. O. 1982; Investigation of the complexity of barley stripe mosaic virus RNAs with recombinant DNA clones. Virology 120:182–193
    [Google Scholar]
  15. Hansen D. R., Van Alfen N. K., Gillies K., Powell W. A. 1985; Naked dsRNA associated with hypovirulence of Endothia parasitica is packaged in fungal vesicles. Journal of General Virology 66:2605–2614
    [Google Scholar]
  16. Joklik W. K. 1983 The Reoviridae New York: Plenum Press;
    [Google Scholar]
  17. Koonin E. V., Choi G. H., Nuss D. L., Shapira R., Carrington J. C. 1991; Evidence for common ancestry of a chestnut blight hypo virulence-associated double-stranded RNA and a group of positive-strand RNA plant viruses. Proceedings of the National Academy of Sciences, U.S.A. 88:10647–10651
    [Google Scholar]
  18. Lefebvre A., Scalla R., Pfeiffer P. 1990; The double-stranded RNA associated with the ‘447’ cytoplasmic male sterility in Vicia faba is packaged together with its replicase in cytoplasmic membranous vesicles. Plant Molecular Biology 14:477–490
    [Google Scholar]
  19. Lockard R. E., Alzner-Deweerd B., Heckman J. E., MacGee J., Tabor M. W., Rajbhandary U. L. 1978; Sequence analysis of 5′ (32P)-labelled mRNA and tRNA using polyacrylamide gel electrophoresis. Nucleic Acids Research 5:37–56
    [Google Scholar]
  20. Morris T. J., Dodds J. A. 1979; Isolation and analysis of doublestranded RNA from virus-infected plant fungal tissue. Phytopathology 68:854–858
    [Google Scholar]
  21. Negruk V. L., Grill L. K., Semancik J. S. 1980; In vitro labelling of viroid RNA for hybridization studies. Journal of Virological Methods 1:229–234
    [Google Scholar]
  22. Nemeroff M. E., Bruenn J. A. 1986; Conservative replication and transcription of Saccharomyces cerevisiae viral double-stranded RNA in vitro. Journal of Virology 57:754–758
    [Google Scholar]
  23. Newhouse J. R., Hoch H. C., MacDonald W. L. 1982; The ultrastructure of Endothia parasitica. Comparison of a virulent with a hypovirulent isolate. Canadian Journal of Botany 61:389–399
    [Google Scholar]
  24. Rae B. R., Hillman B. L., Tartaglia J., Nuss D. L. 1989; Characterization of double-stranded RNA genetic elements associated with biological control of chestnut blight: organization of terminal domains and identification of gene products. EM BO Journal 8:657–663
    [Google Scholar]
  25. Ratti G., Buck K. W. 1978; Semi-conservative transcription in particles of a double-stranded mycovirus. Nucleic Acids Research 5:3843–3854
    [Google Scholar]
  26. Scalla R., Duc G., Rigaud J., Lefebvre A., Meignoz R. 1981; RNA-containing intracellular particles in cytoplasmic male sterile faba bean (Vicia faba L.). Plant Science Letters 22:269–277
    [Google Scholar]
  27. Shapira R., Choi G. H., Nuss D. L. 1991; Virus-like genetic organization and expression strategy for a double-stranded RNA genetic element associated with biological control of chestnut blight. EMBO Journal 10:731–739
    [Google Scholar]
  28. Silberklang M., Prochianz A., Haenni A.-L., Rajbhandary U. L. 1977; Studies on the sequence of the 3′ terminal region of turnip yellow mosaic virus. European Journal of Biochemistry 72:46478
    [Google Scholar]
  29. Sippel A. E. 1973; Purification and characterization of adenosine triphosphate: ribonucleic acid adenyltransferase from Escherichia coli. European Journal of Biochemistry 37:31–40
    [Google Scholar]
  30. Tartaglia J., Paul C. P., Fulbright D. W., Nuss D. L. 1986; Structural properties of double-stranded RNAs associated with biological control of chestnut blight fungus. Proceedings of the National Academy of Sciences, U.S.A. 83:9109–9113
    [Google Scholar]
  31. Turpen T., Garger S. J., Grill L. K. 1988; On the mechanism of cytoplasmic male sterility in the 447 line of Vicia faba. Plant Molecular Biology 10:489–197
    [Google Scholar]
  32. Valverde R., Nameth S., Abdallha O., Al-Musa O., Desjardins P., Dodds J. A. 1990; Indigenous double-stranded RNA from pepper (Capsicum annuum). Plant Science 67:195–201
    [Google Scholar]
  33. Van Alfen N. K., Jaynes R. A., Anagnostakis S. L., Day P. R. 1975; Chestnut blight: biological control by transmissible hypo-virulence in Endothia parasitica. Science 189:890–891
    [Google Scholar]
  34. Wakarchuk D. A., Hamilton R. I. 1985; Cellular doublestranded RNA in Phaseolus vulgaris. Plant Molecular Biology 5:55–63
    [Google Scholar]
  35. Wakarchuk D. A., Hamilton R. I. 1990; Partial nucleotide sequence from enigmatic dsRNA in Phaseolus vulgaris. Plant Molecular Biology 14:637–639
    [Google Scholar]
  36. Williams T. L., Leibowitz M. J. 1987; Conservative mechanism of the in vitro transcription of killer virus of yeast. Virology 158:231–234
    [Google Scholar]
  37. Young E. G., Hanson M. R. 1987; A fused mitochondrial gene associated with cytoplasmic male sterility is developmentally regulated. Cell 50:41–47
    [Google Scholar]
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