1887

Journal of General Virology header logo

Volume 83, Issue 6

Molecular cloning and characterization of cytoplasmic polyhedrosis virus genome segment 9 Free

Abstract

Genome segment 9 of the 11-segment RNA genomes of three cytoplasmic polyhedrosis virus (CPV) isolates from (AmCPV), (AaCPV) and (ApCPV) were converted to cDNA, cloned and sequenced. In each case, this genome segment consists of 1473 nucleotides with one long ORF of 1035 bp and encodes a protein of 345 amino acids, termed NSP38, with a molecular mass of 38 kDa. Secondary structure prediction showed the presence of nine α-helices in the central and terminal domains with localized similarity to RNA-binding motifs of bluetongue virus and infectious bursal disease virus RNA polymerases. Nucleotide sequences were 99·6% identical between these three strains of CPVs, but no similarity was found to any other nucleotide or protein sequence in public databases. The ORF from AmCPV cDNA was expressed as a His-tagged fusion protein in and polyclonal antibody was raised against the purified protein. Immunoblot as well as immunofluorescence analysis with anti-NSP38 antibody showed that the protein was not present in polyhedra or uninfected cells but was present in AmCPV-infected host midgut cells. NSP38 was expressed in insect cells as soluble protein via a baculovirus expression vector and shown to possess the ability to bind poly(rI)–(rC) agarose, which was competitively removed by AmCPV viral RNA. These results indicate that NSP38 is expressed in virus-infected cells as a non-structural protein. By binding to viral RNA, it may play a role in the regulation of genomic RNA function and packaging.

  • Received:
  • Accepted:
  • Published Online:
© Microbiology Society
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-83-6-1483
2002-06-01
2024-04-23
Loading full text...

Full text loading...

/deliver/fulltext/jgv/83/6/0831483a.html?itemId=/content/journal/jgv/10.1099/0022-1317-83-6-1483&mimeType=html&fmt=ahah

References

  1. Altschul S. F., Madden T. L., Schäffer A. A., Zhang J., Zhang Z., Miller W., Lipman D. J. 1997; Gapped blast and psi-blast: a new generation of protein database search programs. Nucleic Acids Research 25:3389–3402
     [Google Scholar]
  2. Arella M., Lavallée C., Belloncik S., Furuichi Y. 1988; Molecular cloning and characterization of cytoplasmic polyhedrosis virus polyhedrin and a viable deletion mutant gene. Journal of Virology 62:211–217
     [Google Scholar]
  3. Ausubel F. M., Brent R., Kingston R. E., Moore D. D., Seidman J. G., Smith J. A. Struhl K. (editors) 1995 Current Protocols in Molecular Biology New York: Wiley;
     [Google Scholar]
  4. Behrens S. E., Tomei L., De Francesco R. 1996; Identification and properties of the RNA-dependent RNA polymerase of hepatitis C virus. EMBO Journal 15:12–22
     [Google Scholar]
  5. Belloncik S. 1989; Cytoplasmic polyhedrosis viruses – Reoviridae. Advances in Virus Research 37:173–209
     [Google Scholar]
  6. Belloncik S., Mori H. 1998; Cypoviruses. In The Insect Viruses pp 337–369 Edited by Miller L. K., Ball L. A. New York: Plenum;
     [Google Scholar]
  7. Belloncik S., Liu J., Su D., Arella M. 1996; Identification and characterization of a new cypovirus, type 14, isolated from Heliothis armigera . Journal of Invertebrate Pathology 67:41–47
     [Google Scholar]
  8. Bittner M., Kupferer P., Morris C. F. 1980; Electrophoretic transfer of proteins and nucleic acids from slab gels to diazobenzyloxymethyl cellulose or nitrocellulose sheets. Analytical Biochemistry 102:459–471
     [Google Scholar]
  9. Blackhall J., Fuentes A., Hansen K., Magnusson G. 1997; Serine protein kinase activity associated with rotavirus phosphoprotein NSP5. Journal of Virology 71:138–144
     [Google Scholar]
  10. Bradford M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72:248–254
     [Google Scholar]
  11. Bruenn J. A. 1991; Relationships among the positive strand and double-strand RNA viruses as viewed through their RNA-dependent RNA polymerases. Nucleic Acids Research 19:217–226
     [Google Scholar]
  12. Caligan J. E., Dunn B. M., Ploegh H. L., Speicher D. W., Wingfield P. T. 1995 Current Protocols in Protein Science New York: Wiley;
     [Google Scholar]
  13. Dolja V. V., Carrington J. C. 1992; Evolution of positive-strand RNA viruses. Seminars in Virology 3:315–326
     [Google Scholar]
  14. Duncan R. 1999; Extensive sequence divergence and phylogenetic relationships between the fusogenic and nonfusogenic orthoreoviruses: a species proposal. Virology 260:316–328
     [Google Scholar]
  15. Echeverry F., Bergeron J., Kaupp W., Guertin C., Arella M. 1997; Sequence analysis and expression of the polyhedrin gene of Choristoneura fumiferana cytoplasmic polyhedrosis virus (CfCPV). Gene 198:399–406
     [Google Scholar]
  16. Estes M. K., Cohen J. 1989; Rotavirus gene structure and function. Microbiological Reviews 53:410–449
     [Google Scholar]
  17. Fabbretti E., Afrikanova I., Vascotto F., Burrone O. R. 1999; Two non-structural rotavirus proteins, NSP2 and NSP5, form viroplasm-like structures in vivo . Journal of General Virology 80:333–339
     [Google Scholar]
  18. Feinberg A. P., Vogelstein B. 1983; A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Analytical Biochemistry 132:6–13
     [Google Scholar]
  19. Fossiez F., Belloncik S., Arella M. 1989; Nucleotide sequence of the polyhedrin gene of Euxoa scandens cytoplasmic polyhedrosis virus (EsCPV. Virology 169:462–465
     [Google Scholar]
  20. Fouillaud M., Morel G. 1994; Characterization of cytoplasmic and nuclear polyhedrosis viruses recovered from the nest of Polistes hebraeus F. (Hymenoptera; Vespidae ). Journal of Invertebrate Pathology 64:89–95
     [Google Scholar]
  21. Galinski M. S., Yu Y., Heminway B. R., Beaudreau G. S. 1994; Analysis of the C-polyhedrin genes from different geographical isolates of a type 5 cytoplasmic polyhedrosis virus. Journal of General Virology 75:1969–1974
     [Google Scholar]
  22. Gillian A. L., Schmechel S. C., Livny J., Schiff L. A., Nibert M. L. 2000; Reovirus protein σNS binds in multiple copies to single-stranded RNA and shares properties with single-stranded DNA binding proteins. Journal of Virology 74:5939–5948
     [Google Scholar]
  23. Hagiwara K., Matsumoto T. 2000; Nucleotide sequences of genome segments 6 and 7 of Bombyx mori cypovirus 1, encoding the viral structural proteins V4 and V5, respectively. Journal of General Virology 81:1143–1147
     [Google Scholar]
  24. Hagiwara K., Tomita M., Nakai K., Kobayashi J., Miyajima S., Yoshimura T. 1998a; Determination of the nucleotide sequence of Bombyx mori cytoplasmic polyhedrosis virus segment 9 and its expression in BmN4 cells. Journal of Virology 72:5762–5768
     [Google Scholar]
  25. Hagiwara K., Tomita M., Kobayashi J., Miyajima S., Yoshimura T. 1998b; Nucleotide sequence of Bombyx mori cytoplasmic polyhedrosis virus segment 8. Biochemical and Biophysical Research Communications 247:549–553
     [Google Scholar]
  26. Hagiwara K., Kobayashi J., Tomito M., Yoshimura T. 2001; Nucleotide sequence of genome segment 5 from Bombyx mori cypovirus 1. Archives of Virology 146:181–187
     [Google Scholar]
  27. Harlow E., Lane D. 1988 Antibodies: a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  28. Hayashi Y., Bird F. T. 1970; The isolation of cytoplasmic polyhedrosis virus from the white-marked tussock moth, Orgyia leucostigma (Smith). Canadian Journal of Microbiology 6:695–701
     [Google Scholar]
  29. Hua J., Chen X., Patton J. T. 1994; Deletion mapping of the rotavirus metalloprotein NS53 (NSP1): the conserved cysteine-rich region is essential for virus-specific RNA binding. Journal of Virology 68:3990–4000
     [Google Scholar]
  30. Hukuhara T., Bonami J. R. 1991; Reoviridae. In Atlas of Invertebrates pp 394–430 Edited by Adams J. R., Bonami J. R. Boca Raton, FL: CRC Press;
     [Google Scholar]
  31. 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. Journal of Virology 75:988–995
     [Google Scholar]
  32. Jolly M. S., Sen S. K., Ahsan M. M. 1974 Tasar Culture Bombay: Ambika Publishers;
     [Google Scholar]
  33. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
     [Google Scholar]
  34. Mori H., Minobe Y., Sasaki T., Kawase S. 1989; Nucleotide sequence of the polyhedrin gene of Bombyx mori cytoplasmic polyhedrosis virus A strain with nuclear localization of polyhedra. Journal of General Virology 70:1885–1888
     [Google Scholar]
  35. Nakashima N., Koizumi M., Watanabe H., Noda H. 1996; Complete nucleotide sequence of the Nilaparvata lugens reovirus: a putative member of the genus Fijivirus . Journal of General Virology 77:139–146
     [Google Scholar]
  36. O’Reilly D. R., Miller L. K., Luckow V. A. 1992 Baculovirus Expression Vectors: a Laboratory Manual New York: W. H. Freeman;
     [Google Scholar]
  37. Payne C. C. 1981; Cytoplasmic polyhedrosis viruses. In Pathogenesis of Invertebrate Microbial Diseases pp 61–100 Edited by Davidson E. W. Totowa, NJ: Allanheld Osmun;
     [Google Scholar]
  38. Payne C. C., Mertens P. P. C. 1983; Cytoplasmic polyhedrosis viruses. In The Reoviridae pp 425–504 Edited by Joklik W. K. New York: Plenum;
     [Google Scholar]
  39. Payne C. C., Rivers C. F. 1976; A provisional classification of cytoplasmic polyhedrosis viruses based on the sizes of the RNA genome segments. Journal of General Virology 33:71–85
     [Google Scholar]
  40. Petrie B. L., Greenberg H. B., Graham D. Y., Estes M. K. 1984; Ultrastructural localization of rotavirus antigens using colloidal gold. Virus Research 1:133–152
     [Google Scholar]
  41. 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]
  42. Poncet D., Aponte C., Cohen J. 1993; Rotavirus protein NSP3 (NS34) is bound to the 3′ end consensus sequence of viral mRNAs in infected cells. Journal of Virology 67:3159–3165
     [Google Scholar]
  43. Qanungo K. R., Kundu S. C., Ghosh A. K. 2000; Characterization of cypovirus isolates from tropical and temperate Indian saturniidae silkworms. Acta Virologica 44:349–357
     [Google Scholar]
  44. Rost B., Sander C. 1994; Combining evolutionary information and neural networks to predict protein secondary structure. Proteins 19:55–72
     [Google Scholar]
  45. Roy P., Marshall J. J., French T. J. 1990; Structure of the bluetongue virus genome and its encoded proteins. Current Topics in Microbiology and Immunology 162:43–87
     [Google Scholar]
  46. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  47. Suzuki N. 1995; Molecular analysis of the rice dwarf virus genome. Seminars in Virology 6:89–95
     [Google Scholar]
  48. Taraporewala Z., Chen D., Patton J. T. 1999; Multimers formed by the rotavirus nonstructural protein NSP2 bind to RNA and have nucleoside triphosphatase activity. Journal of Virology 73:9934–9943
     [Google Scholar]
  49. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences, USA 76:4350–4354
     [Google Scholar]
  50. van Regenmortel M. H. V., Fauquet C. M., Bishop D. H. L., Carstens E. B., Estes M. K., Lemon S. M., Maniloff J., Mayo M. A., McGeoch D. J., Pringle C. R., Wickner R. B. (editors) 2000; Virus Taxonomy. Seventh Report of the International Committee on Taxonomy of Viruses San Diego: Academic Press;
     [Google Scholar]
  51. Yamaguchi T., Ogawa M., Miyoshi M., Inoshima Y., Fukushi H., Hirai K. 1997; Sequence and phylogenetic analyses of highly virulent infectious bursal disease virus. Archives of Virology 142:1441–1458
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-83-6-1483
Loading
Molecular cloning and characterization of Antheraea mylitta cytoplasmic polyhedrosis virus genome segment 9
J. Gen. Virol. 83, 1483 (2002); https://doi.org/10.1099/0022-1317-83-6-1483
/content/journal/jgv/10.1099/0022-1317-83-6-1483
/content/journal/jgv/10.1099/0022-1317-83-6-1483
Loading

Data & Media loading...

Most cited Most Cited RSS feed

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