1887

Abstract

The potyvirus helper component-proteinase (HC-Pro) is a multifunctional protein previously reported to have affinity for polyribonucleotides. To investigate further the ability of HC-Pro to bind nucleic acids, the potato virus Y (PVY) LYE84 isolate HC-Pro gene was amplified, cloned in an expression vector and sequenced. HC-Pro was expressed as a fusion with the maltose-binding protein and purified by affinity chromatography. Electrophoretic mobility-shift assays demonstrated that HC-Pro acts as a sequence non-specific RNA-binding protein and suggest that more than one molecule of protein was bound per molecule of RNA. The HC-Pro RNA-binding activity was stable in 400 m-NaCl and temperature sensitive. The recombinant protein preferentially bound ssRNA over DNA or dsRNA and showed little, if any, affinity for poly(A). The possible implications of the RNA-binding activity of HC-Pro in potyvirus replication and movement are discussed.

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1996-05-01
2024-04-26
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References

  1. Atreya C. D., Pirone T. P. 1993; Mutational analysis of the helper component-proteinase gene of a potyvirus: effects of amino acid substitutions, deletions, and gene replacement on virulence and aphid transmissibility. Proceedings of the National Academy of Sciences, USA 90:11919–11923
    [Google Scholar]
  2. Atreya C. D., Atreya P. L., Thornbury D. W., Pirone T. P. 1992; Site-directed mutations in the potyvirus HC-PRO gene affect helper component activity, virus accumulation, and symptom expression in infected tobacco plants. Virology 191:106–111
    [Google Scholar]
  3. Boyer J. C. 1993 Développement d′ outils moléculaires et cellulaires pour rétude du virus de la mosaïque jaune du navel′, obtention de transcripts infectieux et mise en uvre d’Un systéme protoplastique. Contribution à Lélaboration de stratégies dinterférence avec la réplication virale PhD thesis Université Paris 7;
    [Google Scholar]
  4. Brantley J. D., Hunt A. G. 1993; The N-terminal protein of the polyprotein encoded by the potyvirus tobacco vein mottling virus is an RNA-binding protein. Journal of General Virology 74:1157–1162
    [Google Scholar]
  5. Burd C. G., Dreyfuss G. 1994; Conserved structures and diversity of functions of RNA-binding proteins. Science 265:615–621
    [Google Scholar]
  6. Carrington J. C., Cary S. M., Parks T. D., Dougherty W. G. 1989a; A second proteinase encoded by a plant potyvirus genome. EMBO Journal 8:365–370
    [Google Scholar]
  7. Carrington J. C., Freed D. D., Sanders T. C. 1989b; Auto-catalytic processing of the potyvirus helper component proteinase in Escherichia coli and in vitro. Journal of Virology 63:44595–1463
    [Google Scholar]
  8. Carrington J. C., Freed D. D., Oh C.-S. 1990; Expression of potyviral polyproteins in transgenic plants reveals three proteolytic activities required for complete processing. EMBO Journal 9:1347–1353
    [Google Scholar]
  9. Citovsky V., Knorr D., Schuster G., Zambryski P. 1990; The P30 movement protein of tobacco mosaic virus is a single-strand nucleic acid binding protein. Cell 60:637–647
    [Google Scholar]
  10. Citovsky V., Knorr D., Zambryski P. 1991; Gene I, a potential cell-to-cell movement locus of cauliflower mosaic virus, encodes an RNA-binding protein. Proceedings of the National Academy of Sciences, USA 88:2476–2480
    [Google Scholar]
  11. Citovsky V., Wong M. L., Shaw A. L., Prasad B. V. V., Zambryski P. 1992; Visualization and characterization of tobacco mosaic virus movement protein binding to single-stranded nucleic acids. Plant Cell 4:397–411
    [Google Scholar]
  12. Cronin S., Verchot J., Haldeman-Cahill R., Schaad M. C., Carrington J. C. 1995; Long-distance movement factor: a transport function of the potyvirus helper component proteinase. Plant Cell 7:549–559
    [Google Scholar]
  13. Deom C. M., Lapidot M., Beachy R. N. 1992; Plant virus movement proteins. Cell 69:221–224
    [Google Scholar]
  14. Dolja V. V., Herndon K. L., Pirone T. P., Carrington J. C. 1993; Spontaneous mutagenesis of a plant potyvirus genome after insertion of a foreign gene. Journal of Virology 67:5968–5975
    [Google Scholar]
  15. Dolja V. V., Haldeman R., Robertson N. L., Dougherty W. G., Carrington J. C. 1994; Distinct functions of capsid protein in assembly and movement of tobacco etch potyvirus in plants. EMBO Journal 13:1482–1491
    [Google Scholar]
  16. Dolja V. V., Haldeman-Cahill R., Montgomery A. E., Vandenbosch K. A., Carrington J. C. 1995; Capsid protein determinants involved in cell-to-cell and long distance movement of tobacco etch potyvirus. Virology 206:1007–1016
    [Google Scholar]
  17. Eagle P. A., Klessig D. F. 1992; A zinc-binding motif located between amino acids 273 and 286 in the adenovirus DNA-binding protein is necessary for ssDNA binding. Virology 187:777–787
    [Google Scholar]
  18. Fernández A., Laín S., García J. A. 1995; RNA helicase activity of the plum pox potyvirus CI protein expressed in Escherichia coli. Mapping of an RNA binding domain. Nucleic Acids Research 23:1327–1332
    [Google Scholar]
  19. Fried H. M., Fink G. R. 1978; Electron microscopic heteroduplex analysis of ‘killer’ double-stranded RNA species from yeast. Proceedings of the National Academy of Sciences, USA 75:4224–4228
    [Google Scholar]
  20. Gorelick R. J., Henderson L. E., Hanser J. P., Rein A. 1988; Point mutants of Moloney murine leukemia virus that fail to package viral RNA: evidence for specific RNA recognition by a ‘zinc finger-like’ protein sequence. Proceedings of the National Academy of Sciences, USA 85:8420–8424
    [Google Scholar]
  21. Gramstat A., Courtpozanis A., Rohde W. 1990; The 12 kDa protein of potato virus M displays properties of a nucleic acid-binding regulatory protein. FEBS Letters 276:34–38
    [Google Scholar]
  22. Granier F., Durand-Tardif M., Casse-Delb art F., Lecoq H., Robaglia C. 1993; Mutations in zucchini yellow mosaic virus helper component protein associated with loss of aphid transmissibility. Journal of General Virology 74:2737–2742
    [Google Scholar]
  23. Huet H., Gal-On A., Meir E., Lecoq H., Raccah B. 1994; Mutations in the helper component protease gene of zucchini yellow mosaic virus affect its ability to mediate aphid transmissibility. Journal of General Virology 75:1407–1414
    [Google Scholar]
  24. Klein P. G., Klein R. R., Rodríguez-Cerezo E., Hunt A. G., Shaw J. G. 1994; Mutational analysis of the tobacco vein mottling virus genome. Virology 204:7595–769
    [Google Scholar]
  25. Koonin E. V., Boyko V. P., Dolja V. V. 1991; Small cysteine-rich proteins of different groups of plant RNA viruses are related to different families of nucleic acid-binding proteins. Virology 181:395–398
    [Google Scholar]
  26. Kujawa A. B., Drugeon G., Hulanicka D., Haenni A. L. 1993; Structural requirements for efficient translational frame-shifting in the synthesis of the putative viral RNA-dependent RNA polymerase of potato leafroll virus. Nucleic Acids Research 21:2165–2171
    [Google Scholar]
  27. Maina C. V., Riggs P. D., Grandea A. G. III, Slatko B. E., Moran L. S., Tagliamonte J. A., McReynolds L. A., Guan C. D. 1988; An Escherichia coli vector to express and purify foreign proteins by fusion to and separation from maltose-binding protein. Gene 74:365–373
    [Google Scholar]
  28. Morch M.-D., Boyer J.-C., Haenni A.-L. 1988; Overlapping open reading frames revealed by complete nucleotide sequencing of turnip yellow mosaic virus genomic RNA. Nucleic Acids Research 16:6157–6173
    [Google Scholar]
  29. Osman T. A. M., Hayes R. J., Buck K. W. 1992; Cooperative binding of the red clover necrotic mosaic virus movement protein to single-stranded nucleic acids. Journal of General Virology 73:223–227
    [Google Scholar]
  30. Pirone T. P. 1991; Viral genes and gene products that determine insect transmissibility. Seminars in Virology 2:81–87
    [Google Scholar]
  31. Riechmann J. L., Laín S., García J. A. 1992; Highlights and prospects of potyvirus molecular biology. Journal of General Virology 73:1–16
    [Google Scholar]
  32. Robaglía 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]
  33. Rodríquez P. L., Carrasco L. 1993; Poliovirus protein 2C has ATPase and GTPase activities. Journal of Biological Chemistry 268:8105–8110
    [Google Scholar]
  34. Rouleau M., Smith R. J., Bancroft J. B., Mackie G. A. 1994; Purification, properties, and subcellular localization of foxtail mosaic potexvirus 26-kDa protein. Virology 204:254–265
    [Google Scholar]
  35. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences, USA 74:5463–5467
    [Google Scholar]
  36. Schoumacher F., Erny C., Berna A., Godefroy-Colburn T., Stussi-Garaud C. 1992; Nucleic acid-binding properties of the alfalfa mosaic virus movement protein produced in yeast. Virology 188:896–899
    [Google Scholar]
  37. Soumounou Y., Laliberté J.-F. 1994; Nucleic acid-binding properties of the P1 protein of turnip mosaic potyvirus produced in Escherichia coli . Journal of General Virology 75:2567–2573
    [Google Scholar]
  38. Thomas C. L., Maule A. J. 1995; Identification of the cauliflower mosaic virus movement protein RNA-binding domain. Virology 206:1145–1149
    [Google Scholar]
  39. Thornbury D. W., Hellmann G. M., Rhoads R. E., Pirone T. P. 1985; Purification and characterization of potyvirus helper component. Virology 144:260–267
    [Google Scholar]
  40. Thornbury D. W., Patterson C. A., Dessens J. T., Pirone T. P. 1990; Comparative sequence of the helper component (HC) region of potato virus Y and a HC-defective strain, potato virus C. Virology 178:573–578
    [Google Scholar]
  41. Thornbury D. W., van den Heuvel J. F. J. M., Lesnaw J. A., Pirone T. P. 1993; Expression of potyvirus proteins in insect cells infected with a recombinant baculovirus. Journal of General Virology 74:2731–2735
    [Google Scholar]
  42. Tordo V. M.-J., Chachulska A. M., Fakhfakh H., le Romancer M., Robaglia C., Astier-Manifacier S. 1995; Sequence polymorphism in the 5′NTR and in the P1 coding region of potato virus Y genomic RNA. Journal of General Virology 76:939–949
    [Google Scholar]
  43. Valle R. P. C., Drugeon G., Devignes-Morch M.-D., Legocki A. B., Haenni A.-L. 1992; Codon context effect in virus translational readthrough. A study in vitro of the determinants of TMV and Mo- MuLV amber suppression. FEBS Letters 306:133–139
    [Google Scholar]
  44. Vance V. B., Moore D., Turpen T. H., Brackbr A., Hollowell V. C. 1992; The complete nucleotide sequence of pepper mottle virus genomic RNA: comparison of the encoded polyprotein with those of other sequenced potyviruses. Virology 191:19–30
    [Google Scholar]
  45. Verchot J., Carrington J. C. 1995; Evidence that the potyvirus Pl proteinase functions in trans as an accessory factor for genome amplification. Journal of Virology 69:3668–3674
    [Google Scholar]
  46. Yanisch-Perron C., Vieira J., Messing J. 1985; Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13 mpl8 and pUC19 vectors. Gene 33:103–119
    [Google Scholar]
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