1887

Abstract

ORF 1a of (BYV) encodes the domains of the papain-like proteinase (PCP), methyltransferase (MT) and RNA helicase. BYV cDNA inserts encoding the PCP–MT region were cloned in pGEX vectors next to the glutathione -transferase gene (GST). In a ‘double tag’ construct, the GST–PCP–MT cDNA was flanked by the 3′-terminal six histidine triplets. Following expression in , the fusion proteins were specifically self-cleaved into the GST–PCP and MT fragments. MT-His was purified on Ni–NTA agarose and its N-terminal sequence determined by Edman degradation as GVEEEA, thus providing direct evidence for the Gly/Gly bond cleavage. The GST–PCP fragment purified on glutathione –agarose was used as an immunogen to produce anti-PCP monoclonal antibodies (mAbs). On Western blots of proteins from virus-infected , the mAbs recognized the 66 kDa protein. Immunogold labelling of BYV-infected tissue clearly indicated association of the PCP with the BYV-induced membranous vesicle aggregates, structures related to closterovirus replication.

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2003-08-01
2019-12-16
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References

  1. Agranovsky, A. A. ( 1996; ). The principles of molecular organization, expression and evolution of closteroviruses: over the barriers. Adv Virus Res 47, 119–158.
    [Google Scholar]
  2. Agranovsky, A. A., Koonin, E. V., Boyko, V. P., Maiss, E., Froetschl, R., Lunina, N. A. & Atabekov, J. G. ( 1994; ). Beet yellows closterovirus: complete genome structure and identification of a leader papain-like thiol protease. Virology 198, 311–324.[CrossRef]
    [Google Scholar]
  3. Agranovsky, A. A., Folimonova, S. Y., Folimonov, A. S., Denisenko, O. N. & Zinovkin, R. A. ( 1997; ). The beet yellows closterovirus p65 homologue of HSP70 chaperones has ATPase activity associated with its conserved N-terminal domain but does not interact with unfolded protein chains. J Gen Virol 78, 535–542.
    [Google Scholar]
  4. Ahola, T., Lampio, A., Auvinen, P. & Kääriäinen, L. ( 1999; ). Semliki Forest virus mRNA capping enzyme requires association with anionic membrane phospholipids for activity. EMBO J 18, 3164–3172.[CrossRef]
    [Google Scholar]
  5. Buck, K. ( 1996; ). Comparison of the replication of positive-stranded RNA viruses of plants and animals. Adv Virus Res 47, 159–251.
    [Google Scholar]
  6. Carrington, J. C., Kasschau, K. D. & Johansen, L. K. ( 2001; ). Activation and suppression of RNA silencing by plant viruses. Virology 281, 1–5.[CrossRef]
    [Google Scholar]
  7. Dougherty, W. G. & Semler, B. L. ( 1993; ). Expression of virus-encoded proteinases: functional and structural similarities with cellular enzymes. Microbiol Rev 57, 781–822.
    [Google Scholar]
  8. Erokhina, T. N., Zinovkin, R. A., Vitushkina, M. V., Jelkmann, W. & Agranovsky, A. A. ( 2000; ). Detection of beet yellows closterovirus methyltransferase-like and helicase-like proteins in vivo using monoclonal antibodies. J Gen Virol 81, 597–603.
    [Google Scholar]
  9. Erokhina, T. N., Vitushkina, M. V., Zinovkin, R. A., Lesemann, D. E., Jelkmann, W., Koonin, E. V. & Agranovsky, A. A. ( 2001; ). Ultrastructural localization and epitope mapping of beet yellows closterovirus methyltransferase-like and helicase-like proteins. J Gen Virol 82, 1983–1994.
    [Google Scholar]
  10. Esau, K. & Hoefert, L. L. ( 1971; ). Cytology of beet yellows virus infection in Tetragonia. I. Parenchyma cells in infected leaf. Protoplasma 72, 255–273.[CrossRef]
    [Google Scholar]
  11. Goldbach, R., Le Gall, O. & Wellink, J. ( 1991; ). Alpha-like viruses in plants. Semin Virol 2, 19–25.
    [Google Scholar]
  12. Gorbalenya, A. E., Koonin, E. V. & Lai, M. M. ( 1991; ). Putative papain-related thiol proteases of positive-strand RNA viruses. Identification of rubi- and aphthovirus proteases and delineation of a novel conserved domain associated with proteases of rubi-, alpha- and coronaviruses. FEBS Lett 288, 201–205.[CrossRef]
    [Google Scholar]
  13. Herold, J., Siddell, S. G. & Gorbalenya, A. E. ( 1999; ). A human RNA viral cysteine proteinase that depends upon a unique Zn2+-binding finger connecting the two domains of a papain-like fold. J Biol Chem 274, 14918–14925.[CrossRef]
    [Google Scholar]
  14. Koonin, E. V. & Dolja, V. V. ( 1993; ). Evolution and taxonomy of positive-strand RNA viruses: implications of comparative analysis of amino acid sequences. Crit Rev Biochem Mol Biol 28, 375–430.[CrossRef]
    [Google Scholar]
  15. Lejal, N., Da Costa, B., Huet, J.-C. & Delmas, B. ( 2000; ). Role of Ser-652 and Lys-692 in the protease activity of infectious bursal disease virus VP4 and identification of its substrate cleavage sites. J Gen Virol 81, 983–992.
    [Google Scholar]
  16. Lesemann, D.-E. ( 1988; ). Cytopathology. In The Plant Viruses, vol. 4, pp. 179–235. Edited by R. G. Milne. New York: Plenum.
  17. Lesemann, D.-E. ( 1991; ). Specific cytological alterations in virus-infected plant cells. In Electron Microscopy of Plant Pathogens, pp. 147–159. Edited by K. Mendgen & D.-E. Lesemann. Berlin/Heidelberg/New York: Springer.
  18. Maia, I. G., Haenni, A.-L. & Bernardi, F. ( 1996; ). Potyviral HC-Pro: a multifunctional protein. J Gen Virol 77, 1335–1341.[CrossRef]
    [Google Scholar]
  19. Peng, C. W. & Dolja, V. V. ( 2000; ). Leader proteinase of the beet yellows closterovirus: mutation analysis of the function in genome amplification. J Virol 74, 9766–9770.[CrossRef]
    [Google Scholar]
  20. Peng, C. W., Napuli, A. J. & Dolja, V. V. ( 2003; ). Leader proteinase of beet yellows virus functions in long-distance transport. J Virol 77, 2843–2849.[CrossRef]
    [Google Scholar]
  21. Peremyslov, V. V., Hagiwara, Y. & Dolja, V. V. ( 1998; ). Genes required for replication of the 15·5-kilobase RNA genome of a plant closterovirus. J Virol 72, 5870–5876.
    [Google Scholar]
  22. Rozanov, M. N., Koonin, E. V. & Gorbalenya, A. E. ( 1992; ). Conservation of the putative methyltransferase domain: a hallmark of the ‘Sindbis-like’ supergroup of positive-strand RNA viruses. J Gen Virol 73, 2129–2134.[CrossRef]
    [Google Scholar]
  23. Schwartz, M., Chen, J., Janda, M., Sullivan, M., den Boon, J. & Ahlquist, P. ( 2002; ). A positive-strand RNA virus replication complex parallels form and function of retrovirus capsids. Mol Cell 9, 505–514.[CrossRef]
    [Google Scholar]
  24. Smith, D. B. & Johnson, K. S. ( 1988; ). Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione-S-transferase. Gene 67, 31–40.[CrossRef]
    [Google Scholar]
  25. Strauss, J. H. & Strauss, E. G. ( 1994; ). The alphaviruses: gene expression, replication and evolution. Microbiol Rev 58, 491–562.
    [Google Scholar]
  26. Tijms, M. A., van Dinten, L. C., Gorbalenya, A. E. & Snijder, E. J. ( 2001; ). A zinc finger-containing papain-like protease couples subgenomic mRNA synthesis to genome translation in a positive-stranded RNA virus. Proc Natl Acad Sci U S A 98, 1889–1894.[CrossRef]
    [Google Scholar]
  27. Ziebuhr, J. & Siddell, S. G. ( 1999; ). Processing of the human coronavirus 229E replicase polyproteins by the virus-encoded 3C-like proteinase: identification of proteolytic products and cleavage sites common to pp1a and pp1ab. J Virol 73, 177–185.
    [Google Scholar]
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