Baculovirus expression of the 11 mycoreovirus-1 genome segments and identification of the guanylyltransferase-encoding segment Free

Abstract

The type member (MyRV-1) of a newly described genus, , isolated from a hypovirulent strain 9B21 of the chestnut blight fungus, has a genome composed of 11 dsRNA segments (S1–S11). All of the segments have single ORFs on their capped, positive-sense strands. Infection of insect cells by baculovirus recombinants carrying full-length cDNAs of S1–S11 resulted in overexpression of protein products of the expected sizes, based on their deduced amino acid sequences. This expression system was utilized to identify the S3-encoded protein (VP3) as a guanylyltransferase by an autoguanylylation assay, in which only VP3 was radiolabelled with [-P]GTP. A series of progressive N-terminal and C-terminal deletion mutants was made to localize the autoguanylylation-active site of VP3 to aa residues 133–667. Within this region, a sequence stretch (aa 170–250) with relatively high sequence similarity to homologues of two other mycoreoviruses and two coltiviruses was identified. Site-directed mutagenesis of conserved aa residues revealed that H233, H242, Y243, F244 and F246, but not K172 or K202, play critical roles in guanylyltransferase activities. Together with broader sequence alignments of ‘turreted’ reoviruses, these results supported the a/vxxHxHyf/lvf motif, originally noted for orthoreovirus and aquareoviruses, as an active site for guanylyltransferases of viruses within the , , , , , and genera, as well as for the proposed genus.

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2007-01-01
2024-03-29
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References

  1. Asamizu T., Summers D., Motika M. B., Anzola J. V., Nuss D. L. 1985; Molecular cloning and characterization of the genome of wound tumor virus: a tumor-inducing plant reovirus. Virology 144:398–409 [CrossRef]
    [Google Scholar]
  2. Attoui H., Mohd Jaafar F., Biagini P., Cantaloube J. F., de Micco P., Murphy F. A., de Lamballerie X. 2002; Genus Coltivirus (family Reoviridae ): genomic and morphologic characterization of Old World and New World viruses. Arch Virol 147:533–561 [CrossRef]
    [Google Scholar]
  3. Attoui H., Mohd Jaafar F., Belhouchet M., Biagini P., Cantaloube J. F., de Micco P., de Lamballerie X. 2005; Expansion of family Reoviridae to include nine-segmented dsRNA viruses: isolation and characterization of a new virus designated Aedes pseudoscutellaris reovirus assigned to a proposed genus ( Dinovernavirus ). Virology 343:212–223 [CrossRef]
    [Google Scholar]
  4. Attoui H., Mohd Jaafar F., Belhouchet M., de Micco P., de Lamballerie X., Brussaard C. P. 2006; Micromonas pusilla reovirus : a new member of the family Reoviridae assigned to a novel proposed genus ( Mimoreovirus ). J Gen Virol 87:1375–1383 [CrossRef]
    [Google Scholar]
  5. Churchill A. C. L., Ciufetti L. M., Hansen D. R., Van Etten H. D., Van Alfen N. K. 1990; Transformation of the fungal pathogen Cryphonectria parasitica with a variety of heterologous plasmids. Curr Genet 17:25–31 [CrossRef]
    [Google Scholar]
  6. Cleveland D. R., Zarbl H., Millward S. 1986; Reovirus guanylyltransferase is L2 gene product lambda 2. J Virol 60:307–311
    [Google Scholar]
  7. Enebak S. A. 1992; Characterization of dsRNA-containing strains of Cryphonectria parasitica recovered from the central Appalachian . PhD thesis West Virginia University;
  8. Estes M. K. 2001; Rotaviruses and their replication. In Fields Virology , 4th edn. vol 2 pp  1747–1785 Edited by Knipe D. M., Howley P. M. Philadelphia: Lippincott, Williams & Wilkins;
    [Google Scholar]
  9. Fausnaugh J., Shatkin A. J. 1990; Active site localization in a viral mRNA capping enzyme. J Biol Chem 265:7669–7672
    [Google Scholar]
  10. Furuichi Y., Shatkin A. J. 2000; Viral and cellular capping: past and prospects. Adv Virus Res 55:135–184
    [Google Scholar]
  11. Hill C. L., Booth T. F., Prasad B. V., Grimes J. M., Mertens P. P., Sutton G. C., Stuart D. I. 1999; The structure of a cypovirus and the functional organization of dsRNA viruses. Nat Struct Biol 6:565–568 [CrossRef]
    [Google Scholar]
  12. Hillman B. I., Suzuki N. 2004; Viruses in the chestnut blight fungus. Adv Virus Res 63:423–472
    [Google Scholar]
  13. Hillman B. I., Shapira R., Nuss D. L. 1990; Hypovirulence-associated suppression of host functions in Cryphonectria parasitica can be partially relieved by high light intensity. Phytopathology 80:950–956 [CrossRef]
    [Google Scholar]
  14. Hillman B. I., Supyani S., Kondo H., Suzuki N. 2004; A reovirus of the fungus Cryphonectria parasitica that is infectious as particles and related to the Coltivirus genus of animal pathogen. J Virol 78:892–898 [CrossRef]
    [Google Scholar]
  15. Hsiao J., Martinez-Costas J., Benavente J., Vakharia V. N. 2002; Cloning, expression, and characterization of avian reovirus guanylyltransferase. Virology 296:288–299 [CrossRef]
    [Google Scholar]
  16. Huang Y. L., Han Y. T., Chang Y. T., Hsu Y. H., Meng M. 2004; Critical residues for GTP methylation and formation of the covalent m7GMP–enzyme intermediate in the capping enzyme domain of bamboo mosaic virus. J Virol 78:1271–1280 [CrossRef]
    [Google Scholar]
  17. Ikeda K., Nagaoka S., Winkler S., Kotani K., Yagi H., Nakanishi K., Miyajima S., Kobayashi J., Mori H. 2001; Molecular characterization of Bombyx mori cytoplasmic polyhedrosis virus genome segment 4. J Virol 75:988–995 [CrossRef]
    [Google Scholar]
  18. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685 [CrossRef]
    [Google Scholar]
  19. Lawton J. A., Estes M. K., Prasad B. V. 2000; Mechanism of genome transcription in segmented dsRNA viruses. Adv Virus Res 55:185–229
    [Google Scholar]
  20. Le Blois H., French T., Mertens P. P. C., Burroughs J. N., Roy P. 1992; The expressed VP4 protein of bluetongue virus binds GTP and is the candidate guanylyl transferase of the virus. Virology 189:757–761 [CrossRef]
    [Google Scholar]
  21. Liu M., Mattion N. M., Estes M. K. 1992; Rotavirus VP3 expressed in insect cells possesses guanylyltransferase activity. Virology 188:77–84 [CrossRef]
    [Google Scholar]
  22. Luongo C. L. 2002; Mutational analysis of a mammalian reovirus mRNA capping enzyme. Biochem Biophys Res Commun 291:932–938 [CrossRef]
    [Google Scholar]
  23. Matsuura Y., Possee R. D., Overton H. A., Bishop D. H. L. 1987; Baculovirus expression vectors: the requirements for high level expression of proteins, including glycoproteins. J Gen Virol 68:1233–1250 [CrossRef]
    [Google Scholar]
  24. Mertens P. P. C., Attoui H., Duncan R., Dermody T. S. 2005a; Reoviridae . In Virus Taxonomy: Eighth Report of the International Committee on Taxonomy of Viruses pp  447–454 Edited by Fauquet C. M., Mayo M. A., Maniloff J., Desselberger U., Ball L. A. London: Elsevier/Academic Press;
    [Google Scholar]
  25. Mertens P. P. C., Wei C. Z., Hillman B. I. 2005b; Genus Mycoreovirus . In Virus Taxonomy: Eighth Report of the International Committee on Taxonomy of Viruses pp  556–560 Edited by Fauquet C. M., Mayo M. A., Maniloff J., Desselberger U., Ball L. A. London: Elsevier/Academic Press;
    [Google Scholar]
  26. Mizumoto K., Kaziro Y. 1987; Messenger RNA capping enzymes from eukaryotic cells. Prog Nucleic Acid Res Mol Biol 34:1–28
    [Google Scholar]
  27. Mohd Jaafar F., Attoui H., Mertens P. P. C., de Micco P., de Lamballerie X. 2005; Structural organization of an encephalitic human isolate of Banna virus (genus Seadornavirus , family Reoviridae ). J Gen Virol 86:1147–1157 [CrossRef]
    [Google Scholar]
  28. Nibert M. L., Schiff L. A. 2001; Reoviruses and their replication. In Fields Virology , 4th edn. vol 2 pp  1679–1729 Edited by Knipe D. M., Howley P. M. Philadelphia: Lippincott Williams & Wilkins;
    [Google Scholar]
  29. Nibert M. L., Kim J. 2004; Conserved sequence motifs for nucleotide triphosphate binding unique to turreted Reoviridae members and coltiviruses. J Virol 78:5528–5530 [CrossRef]
    [Google Scholar]
  30. Patton J. T., Jones M. T., Kalbach A. N., He Y. W., Xiaobo J. 1997; Rotavirus RNA polymerase requires the core shell protein to synthesize the double-stranded RNA genome. J Virol 71:9618–9626
    [Google Scholar]
  31. Qiu T., Luongo C. L. 2003; Identification of two histidines necessary for reovirus mRNA guanylyltransferase activity. Virology 316:313–324 [CrossRef]
    [Google Scholar]
  32. Ramadevi N., Burroughs J. N., Mertens P. P. C., Jones I. M., Roy P. 1998; Capping and methylation of mRNA by purified recombinant VP4 protein of bluetongue virus. Proc Natl Acad Sci U S A 95:13537–13542 [CrossRef]
    [Google Scholar]
  33. Reinisch K. M., Nibert M. L., Harrison S. C. 2000; Structure of the reovirus core at 3.6 Å resolution. Nature 404:960–967 [CrossRef]
    [Google Scholar]
  34. Roy P. 2001; Orbiviruses. In Fields Virology . , 4th edn. vol 2 pp  1835–1869 Edited by Knipe D. M., Howley P. M. Philadelphia: Lippincott Williams & Wilkins;
  35. Sambrook J., Russell D. W. 2001 Molecular Cloning: a Laboratory Manual , 3rd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  36. Shuman S., Hurwitz J. 1981; Mechanism of mRNA capping by vaccinia virus guanylyltransferase: characterization of an enzyme–guanylate intermediate. Proc Natl Acad Sci U S A 78:187–191 [CrossRef]
    [Google Scholar]
  37. Suzuki N., Sugawara M., Kusano T., Mori H., Matsuura Y. 1994; Immunodetection of rice dwarf phytoreoviral proteins in both insect and plant hosts. Virology 202:41–48 [CrossRef]
    [Google Scholar]
  38. Suzuki N., Kusano T., Matsuura Y., Omura T. 1996; Novel NTP binding property of rice dwarf phytoreovirus minor core protein P5. Virology 219:471–474 [CrossRef]
    [Google Scholar]
  39. Suzuki N., Supyani S., Maruyama K., Hillman B. I. 2004; Complete genome sequence of Mycoreovirus 1 /Cp9B21, a member of a new genus within the family Reoviridae , from the chestnut blight fungus Cryphonectria parasitica . J Gen Virol 85:3437–3448 [CrossRef]
    [Google Scholar]
  40. Thompson J. D., Gibson T. J., Plewniak F., Jeanmougin F., Higgins D. G. 1997; The clustal_x windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25:4876–4882 [CrossRef]
    [Google Scholar]
  41. Wei C. Z., Osaki H., Iwanami T., Matsumoto N., Ohtsu Y. 2004; Complete nucleotide sequences of genome segments 1 and 3 of Rosellinia anti-rot virus in the family Reoviridae . Arch Virol 149:773–777 [CrossRef]
    [Google Scholar]
  42. Wei T., Shimizu T., Hagiwara K., Kikuchi A., Moriyasu Y., Suzuki N., Chen H., Omura T. 2006; Pns12 protein of Rice dwarf virus is essential for formation of viroplasms and nucleation of viral-assembly complexes. J Gen Virol 87:429–438 [CrossRef]
    [Google Scholar]
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