1887

Abstract

The complete nucleotide sequence of the RNA genome of papaya ringspot virus (PRSV) was determined from four overlapping cDNA clones and by direct sequencing of viral RNA. The genomic RNA is 10326 nucleotides in length, excluding the poly(A) tract, and contains one large open reading frame that starts at nucleotide positions 86 to 88 and ends at positions 10118 to 10120, encoding a polyprotein of 3344 amino acids. The highly conserved sequence AAAUAAAANANCUCAACACAACAUA at the 5′ end of the RNA of PRSV and those of the other five reported potyviruses shows 80% similarity, suggesting that this region may play a common important role for potyvirus replication. Two cleavage sites of the polyprotein were determined by amino acid sequencing of the N termini of helper component (HC-Pro, amorphous inclusion) and cylindrical inclusion (CI) proteins. Other cleavage sites were predicted by analogy with the other potyviruses. The genetic organization of PRSV is similar to that of the other potyviruses except that the first protein processed from the N terminus of the polyprotein (NT protein) has an of 63K, 18K to 34K larger than those of the other potyviruses. The cleavage site for liberating the N terminus of the HC-Pro protein was found at the same location downstream from the consensus sequence FI(V)VRG as that reported for tobacco vein mottling virus. The NT protein of potyviruses is the most variable and may be considered important for identification of individual potyviruses. The most conserved protein of potyviruses appears to be the NIb protein, the putative polymerase for the replication of the potyviral RNA. The genetic organization of PRSV RNA is tentatively proposed to be VPg-5′ leader-63K NT-52K HC-Pro-46K-72K CI-6K-48K NIa-59K NIb-35K coat protein-3′ noncoding region-poly(A) tract.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-73-10-2531
1992-10-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/jgv/73/10/JV0730102531.html?itemId=/content/journal/jgv/10.1099/0022-1317-73-10-2531&mimeType=html&fmt=ahah

References

  1. Allison R. F., Johnston R. E., Dougherty W. G. 1986; The nucleotide sequence of the coding region of tobacco etch virus genomic RNA: evidence for the synthesis of a single polyprotein. Virology 154:9–20
    [Google Scholar]
  2. Carrington J. C., Dougherty W. G. 1987a; Small nuclear inclusion protein encoded by a plant potyvirus genome is a protease. Journal of Virology 61:2540–2548
    [Google Scholar]
  3. Carrington J. C., Dougherty W. G. 1987b; Processing of the tobacco etch virus 49K protease requires autoproteolysis. Virology 160:355–362
    [Google Scholar]
  4. Carrington J. C., Cary S. M., Parks T. D., Dougherty W. G. 1989; A second proteinase encoded by a plant potyvirus. EMBO Journal 8:365370
    [Google Scholar]
  5. De La Rosa M., Lastra R. 1983; Purification and partial characterization of papaya ringspot virus. Phytopathologische Zeitschrift 106:329–336
    [Google Scholar]
  6. De Mejia M. V. G., Hiebert E., Purcifull D. E. 1985a; Isolation and partial characterization of the amorphous cytoplasmic inclusions associated with infections caused by two potyviruses. Virology 142:24–33
    [Google Scholar]
  7. De Mejia M. V. G., Hiebert E., Purcifull D. E., Thornbury D. W., Pirone T. P. 1985b; Identification of potyviral amorphous inclusion protein as a nonstructural, virus-specific protein related to helper component. Virology 142:34–43
    [Google Scholar]
  8. Dinant S., Lot H., Albouy J., Kuziak C., Meyer M., Astier-Manifacier S. 1991; Nucleotide sequence of the 3′ terminal region of lettuce mosaic potyvirus RNA shows a Gln/Val dipeptide at the cleavage site between the polymerase and the coat protein. Archives of Virology 116:235–252
    [Google Scholar]
  9. Domier L. L., Franklin K. M., Shahabuddin M., Hellmann G. M., Overmeyer J. H., Hiremath S. T., Siaw M. F. E., Lomonossoff G. P., Shaw J. G., Rhoads R. E. 1986; The nucleotide sequence of tobacco vein mottling virus RNA. Nucleic Acids Research 14:5417–5430
    [Google Scholar]
  10. Domier L. L., Shaw J. G., Rhoads R. E. 1987; Potyviral proteins share amino acid sequence homology with picoma-, como-, and caulimoviral proteins. Virology 158:20–27
    [Google Scholar]
  11. Dougherty W. G., Carrington J. C. 1988; Expression and function of potyviral gene products. Annual Review of Phytopathology 26:123–143
    [Google Scholar]
  12. Dougherty W. G., Parks T. D. 1991; Post-translational processing of the tobacco etch virus 49-kDa small nuclear inclusion polyprotein: identification of an internal cleavage site and delimitation of VPg and proteinase domains. Virology 183:449–456
    [Google Scholar]
  13. Dougherty W. G., Cary S. M., Parks T. D. 1989a; Molecular genetic analysis of a plant virus polyprotein cleavage site: a model. Virology 171:356–364
    [Google Scholar]
  14. Dougherty W. G., Parks T. D., Cary S. M., Bazan J. F., Fletterick R. J. 1989b; Characterization of the catalytic residues of the tobacco etch virus 49-kDa proteinase. Virology 172:302–310
    [Google Scholar]
  15. Ghabrial S. A., Smith H. A., Parks T. D., Dougherty W. G. 1990; Molecular genetic analyses of the soybean mosaic virus NIa proteinase. Journal of General Virology 71:1921–1927
    [Google Scholar]
  16. Gonsalves D., Ishii M. 1980; Purification and serology of papaya ringspot virus. Phytopathology 70:1028–1032
    [Google Scholar]
  17. Gorbalenya A. E., Koonin E. V., Doncheko A. P., Blinov V. M. 1989; A novel superfamily of nucleoside triphosphatebinding motif containing proteins which are probably involved in duplex unwinding in DNA and RNA replication and recombination. FEBS Letters 235:16–24
    [Google Scholar]
  18. Gubler U., Hoffman B. J. 1983; A simple and very efficient method for generating cDNA libraries. Gene 25:263–269
    [Google Scholar]
  19. Guo L. H., Wu R. 1982; New rapid methods for DNA sequencing based on exonuclease III digestion followed by repair synthesis. Nucleic Acids Research 10:2065–2084
    [Google Scholar]
  20. Hari V. 1981; The RNA of tobacco etch virus: further characterization and detection of protein linked to RNA. Virology 112:391–399
    [Google Scholar]
  21. Henikoff S. 1984; Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28:351–359
    [Google Scholar]
  22. Huynh T. V., Young R. A., Davis R. W. 1985; Constructing and screening cDNA libraries in λ gt10 and λ gt11. In DNA Cloning: A Practical Approach vol 1 pp. 49–79 Edited by Glover D. M. Oxford: IRL Press;
    [Google Scholar]
  23. Johansen E., Rasmussen O. F., Heide M., Borkhardt B. 1991; The complete nucleotide sequence of pea seed-borne mosaic virus RNA. Journal of General Virology 72:2625–2632
    [Google Scholar]
  24. Kozak M. 1989; The scanning model for translation: an update. Journal of Cell Biology 108:229–241
    [Google Scholar]
  25. Laίn S., Riechmann J. L., Garcίa J. A. 1990; RNA helicase: a novel activity associated with a protein encoded by a positive strand RNA virus. Nucleic Acids Research 18:7003–7006
    [Google Scholar]
  26. Laίn S., Martίn M. T., Riechmann J. L., Garcίa J. A. 1991; Novel catalytic activity associated with positive-strand RNA virus infection: nucleic acid-stimulated ATPase activity of the plum pox potyvirus helicase like protein. Journal of Virology 65:1–6
    [Google Scholar]
  27. Lutcke H. A., Chow K. C., Mickel F. S., Moss K. A., Kern H. F., Scheele G. A. 1987; Selection of AUG initiation codons differs in plants and animals. EMBO Journal 6:43–48
    [Google Scholar]
  28. Maiss E., Timpe U., Brisske A., Jelkmann W., Casper R., Himmler G., Mattanovich D., Katinger H. W. D. 1989; The complete nucleotide sequence of plum pox virus RNA. Journal of General Virology 70:513–524
    [Google Scholar]
  29. Martelli G. P., Russo M. 1976; Unusual cytoplasmic inclusions induced by watermelon mosaic virus. Virology 72:352–362
    [Google Scholar]
  30. Mavankal G., Rhoads R. E. 1991; In vitro cleavage at or near the N-terminus of the helper component protein in the tobacco vein mottling virus polyprotein. Virology 185:721–731
    [Google Scholar]
  31. Murphy J. F., Rhoads R. E., Hunt A. G., Shaw J. G. 1990; The VPg of tobacco etch virus RNA is the 49 kDa proteinase or the N-terminal 24-kDa part of the proteinase. Virology 178:285–288
    [Google Scholar]
  32. Nagel J., Hiebert E. 1985; Complementary DNA cloning and expressing of the papaya ringspot potyvirus sequences encoding capsid protein and a nuclear inclusion-like protein in Escherichia coli . Virology 143:435–441
    [Google Scholar]
  33. Oh C.-S., Carrington J. C. 1989; Identification of essential residues in potyvirus proteinase HC-Pro by site-directed mutagenesis. Virology 173:692–699
    [Google Scholar]
  34. Poch O., Sauvaget I., Delarue M., Tordo N. 1989; Identification of four conserved motifs among the RNA-dependent polymerase encoding elements. EMBO Journal 8:3867–3874
    [Google Scholar]
  35. Prowidenti R., Gonsalves D. 1982; Resistance to papaya ringspot virus in Cucumis metuliferus and its relationship to resistance to watermelon mosaic virus 1. Journal of Heredity 73:239–240
    [Google Scholar]
  36. Purcifull D. E., Edwardson J. R. 1967; Watermelon mosaic virus: tubular inclusion in pumpkin leaves and aggregates in leaf extracts. Virology 32:393–401
    [Google Scholar]
  37. Purcifull D. E., Hiebert E. 1979; Serological distinction of watermelon mosaic virus isolates. Phytopathology 69:112–116
    [Google Scholar]
  38. Purcifull D. E., Edwardson J. R., Hiebert E., Gonsalves D. 1984; Papaya ringspot virus. CMI/AAB Descriptions of Plant Viruses no. 292
    [Google Scholar]
  39. Putney S. D., Benkovic S. J., Schimmel P. 1981; A DNA fragment with an α-phosphorothioate nucleotide at one end is asymmetrically blocked from digestion by exonuclease III and can be replicated in vivo . Proceedings of the National Academy of Sciences, U.S.A. 78:7350–7354
    [Google Scholar]
  40. Quemada H., L’Hostis B., Gonsalves D., Reardon I. M., Heinrikson R., Hiebert E. L., Sieu L. C., Slightom J. L. 1990; The nucleotide sequences of the 3′-terminal regions of papaya ringspot virus strains W and P. Journal of General Virology 71:203–210
    [Google Scholar]
  41. Quiot-Douine L., Purcifull D. E., De Mejia M. V. G. 1986; Serological relationships and in vitro translation of an antigenically distinct strain of papaya ringspot virus. Phytopathology 76:346–351
    [Google Scholar]
  42. Riechmann J. L., Laίn S., Garcίa J. A. 1989; The genome-linked protein and 5′ end RNA sequence of plum pox potyvirus. Journal of General Virology 70:2785–2789
    [Google Scholar]
  43. Robaglia C., Durand-Tardif M., Tronchet M., Boudazin G., Astier-Manifacier S., Casse-Delbart F. 1989; Nucleotide sequence of potato virus Y (N strain) genomic RNA. Journal of General Virology 70:935–947
    [Google Scholar]
  44. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: A Laboratory Manual 2nd edn New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  45. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy Science, U.S.A. 74:5463–5467
    [Google Scholar]
  46. Shuxla D. D., Ward C. W. 1989; Structure of potyvirus coat proteins and its application in the taxonomy of the potyvirus group. Advances in Virus Research 36:273–314
    [Google Scholar]
  47. Siaw M. F. E., Shahabuddin M., Ballard S., Shaw J. G., Rhoads R. E. 1985; Identification of a protein covalently linked to the 5′ terminus of tobacco vein mottling virus RNA. Virology 142:134–143
    [Google Scholar]
  48. Verchot J., Koonin E. V., Carrington J. C. 1991; The 35-kDa protein from the N-terminus of the potyviral polyprotein functions as a third virus-encoded proteinase. Virology 185:527–535
    [Google Scholar]
  49. Wang C. H., Yeh S.-D. 1992; Nucleotide sequence comparison of the 3′-terminal region of severe, mild, and non-papaya infecting strains of papaya ringspot virus. Archives of Virology (in press)
    [Google Scholar]
  50. Wang H.-L., Yeh S.-D., Chiu R.-J., Gonsalves D. 1987; Effectiveness of cross-protection by mild mutants of papaya ringspot virus for control of ringspot disease of papaya in Taiwan. Plant Disease 71:491–497
    [Google Scholar]
  51. Yeh S.-D., Bih F. Y. 1989; Comparative studies on in vitro translation of a severe strain and a mild strain of papaya ringspot virus. Plant Protection Bulletin (Taiwan, R,. O,. C.) 31:276–289
    [Google Scholar]
  52. Yeh S.-D., Gonsalves D. 1984a; Evaluation of induced mutants of papaya ringspot virus for control by cross protection. Phytopathology 74:1086–1091
    [Google Scholar]
  53. Yeh S.-D., Gonsalves D. 1984b; Purification and immunological analyses of cylindrical-inclusion protein induced by papaya ringspot virus and watermelon mosaic virus 1. Phytopathology 74:1273–1278
    [Google Scholar]
  54. Yeh S.-D., Gonsalves D. 1985; Translation of papaya ringspot virus RNA in vitro: detection of a possible polyprotein that is processed for capsid protein, cylindrical-inclusion protein, and amorphous-inclusion protein. Virology 143:260–271
    [Google Scholar]
  55. Yeh S.-D., Gonsalves D., Prowidenti R. 1984; Comparative studies on host range and serology of papaya ringspot virus and watermelon mosaic virus 1. Phytopathology 74:1081–1085
    [Google Scholar]
  56. Yeh S.-D., Gonsalves D., Wang H. -L., Namba R., Chiu R. -J. 1988; Control of papaya ringspot virus by cross protection. Plant Disease 72:375–380
    [Google Scholar]
  57. Youvan D. C., Hearst J. E. 1981; A sequence from Drosophila melanogaster 18S rRNA bearing the conserved hypermodified nucleotide: analysis by reverse transcription and high-performance liquid chromatography. Nucleic Acids Research 9:1723–1741
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-73-10-2531
Loading
/content/journal/jgv/10.1099/0022-1317-73-10-2531
Loading

Data & Media loading...

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