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. 1997 Jul;71(7):5040–5050. doi: 10.1128/jvi.71.7.5040-5050.1997

Identification and characterization of a baculovirus structural protein, VP1054, required for nucleocapsid formation.

J Olszewski 1, L K Miller 1
PMCID: PMC191737  PMID: 9188569

Abstract

The defect in a temperature-sensitive mutant of Autographa californica nuclear polyhedrosis virus, tsN1054, was mapped and characterized. At the nonpermissive temperature of 33 degrees C, this mutant fails to form plaques upon infection of Sf-21 cultured insect cells; infection is limited to a single cell, even though the infection proceeds through the very late phase. Marker rescue mapping and DNA sequencing identified the gene, ORF 54, which was altered by a single nucleotide substitution in tsN1054. Transcriptional analysis of the ORF 54 region identified multicistronic RNAs, from early to very late times of infection, that potentially encode the ORF 54 gene product. Polyclonal antiserum raised to a TrpE-VP1054 fusion protein recognized a 42-kDa late protein, VP1054, in infected-cell lysates. VP1054 was found to be a component of both budded virus and occlusion-derived virions. The level of VP1054 was dramatically reduced in tsN1054-infected Sf-21 cells propagated at 33 degrees C, and electron microscopic analysis of these cells showed that nucleocapsids failed to form in the nuclei of these infected cells. Instead, novel round, electron-dense bodies were found associated with the virogenic stroma in tsN1054-infected cells. Therefore, VP1054 is a virus structural protein required for nucleocapsid assembly.

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Selected References

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  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Ayres M. D., Howard S. C., Kuzio J., Lopez-Ferber M., Possee R. D. The complete DNA sequence of Autographa californica nuclear polyhedrosis virus. Virology. 1994 Aug 1;202(2):586–605. doi: 10.1006/viro.1994.1380. [DOI] [PubMed] [Google Scholar]
  3. Blissard G. W., Quant-Russell R. L., Rohrmann G. F., Beaudreau G. S. Nucleotide sequence, transcriptional mapping, and temporal expression of the gene encoding p39, a major structural protein of the multicapsid nuclear polyhedrosis virus of Orgyia pseudotsugata. Virology. 1989 Feb;168(2):354–362. doi: 10.1016/0042-6822(89)90276-6. [DOI] [PubMed] [Google Scholar]
  4. Blissard G. W., Rohrmann G. F. Baculovirus diversity and molecular biology. Annu Rev Entomol. 1990;35:127–155. doi: 10.1146/annurev.en.35.010190.001015. [DOI] [PubMed] [Google Scholar]
  5. Braunagel S. C., Elton D. M., Ma H., Summers M. D. Identification and analysis of an Autographa californica nuclear polyhedrosis virus structural protein of the occlusion-derived virus envelope: ODV-E56. Virology. 1996 Mar 1;217(1):97–110. doi: 10.1006/viro.1996.0097. [DOI] [PubMed] [Google Scholar]
  6. Braunagel S. C., He H., Ramamurthy P., Summers M. D. Transcription, translation, and cellular localization of three Autographa californica nuclear polyhedrosis virus structural proteins: ODV-E18, ODV-E35, and ODV-EC27. Virology. 1996 Aug 1;222(1):100–114. doi: 10.1006/viro.1996.0401. [DOI] [PubMed] [Google Scholar]
  7. Braunagel S. C., Summers M. D. Autographa californica nuclear polyhedrosis virus, PDV, and ECV viral envelopes and nucleocapsids: structural proteins, antigens, lipid and fatty acid profiles. Virology. 1994 Jul;202(1):315–328. doi: 10.1006/viro.1994.1348. [DOI] [PubMed] [Google Scholar]
  8. Goh D. K. Identification and immunocharacterization of a 25K structural protein in Autographa californica nuclear polyhedrosis virus (AcMNPV). Virus Res. 1993 May;28(2):141–152. doi: 10.1016/0168-1702(93)90132-7. [DOI] [PubMed] [Google Scholar]
  9. Harrap K. A. The structure of nuclear polyhedrosis viruses. 3. Virus assembly. Virology. 1972 Oct;50(1):133–139. doi: 10.1016/0042-6822(72)90353-4. [DOI] [PubMed] [Google Scholar]
  10. Hong T., Braunagel S. C., Summers M. D. Transcription, translation, and cellular localization of PDV-E66: a structural protein of the PDV envelope of Autographa californica nuclear polyhedrosis virus. Virology. 1994 Oct;204(1):210–222. doi: 10.1006/viro.1994.1525. [DOI] [PubMed] [Google Scholar]
  11. Keddie B. A., Aponte G. W., Volkman L. E. The pathway of infection of Autographa californica nuclear polyhedrosis virus in an insect host. Science. 1989 Mar 31;243(4899):1728–1730. doi: 10.1126/science.2648574. [DOI] [PubMed] [Google Scholar]
  12. Koerner T. J., Hill J. E., Myers A. M., Tzagoloff A. High-expression vectors with multiple cloning sites for construction of trpE fusion genes: pATH vectors. Methods Enzymol. 1991;194:477–490. doi: 10.1016/0076-6879(91)94036-c. [DOI] [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Lee H. H., Miller L. K. Isolation, Complementation, and Initial Characterization of Temperature-Sensitive Mutants of the Baculovirus Autographa californica Nuclear Polyhedrosis Virus. J Virol. 1979 Jul;31(1):240–252. doi: 10.1128/jvi.31.1.240-252.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Li Y., Miller L. K. Expression and localization of a baculovirus protein phosphatase. J Gen Virol. 1995 Dec;76(Pt 12):2941–2948. doi: 10.1099/0022-1317-76-12-2941. [DOI] [PubMed] [Google Scholar]
  16. Li Y., Miller L. K. Properties of a baculovirus mutant defective in the protein phosphatase gene. J Virol. 1995 Jul;69(7):4533–4537. doi: 10.1128/jvi.69.7.4533-4537.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lu A., Carstens E. B. Nucleotide sequence of a gene essential for viral DNA replication in the baculovirus Autographa californica nuclear polyhedrosis virus. Virology. 1991 Mar;181(1):336–347. doi: 10.1016/0042-6822(91)90500-b. [DOI] [PubMed] [Google Scholar]
  18. Lu A., Miller L. K. Identification of three late expression factor genes within the 33.8- to 43.4-map-unit region of Autographa californica nuclear polyhedrosis virus. J Virol. 1994 Oct;68(10):6710–6718. doi: 10.1128/jvi.68.10.6710-6718.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Maeda S., Kamita S. G., Kataoka H. The basic DNA-binding protein of Bombyx mori nuclear polyhedrosis virus: the existence of an additional arginine repeat. Virology. 1991 Feb;180(2):807–810. doi: 10.1016/0042-6822(91)90096-t. [DOI] [PubMed] [Google Scholar]
  20. McMaster G. K., Carmichael G. G. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4835–4838. doi: 10.1073/pnas.74.11.4835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Monsma S. A., Oomens A. G., Blissard G. W. The GP64 envelope fusion protein is an essential baculovirus protein required for cell-to-cell transmission of infection. J Virol. 1996 Jul;70(7):4607–4616. doi: 10.1128/jvi.70.7.4607-4616.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Müller R., Pearson M. N., Russell R. L., Rohrmann G. F. A capsid-associated protein of the multicapsid nuclear polyhedrosis virus of Orgyia pseudotsugata: genetic location, sequence, transcriptional mapping, and immunocytochemical characterization. Virology. 1990 May;176(1):133–144. doi: 10.1016/0042-6822(90)90238-m. [DOI] [PubMed] [Google Scholar]
  23. Passarelli A. L., Miller L. K. Three baculovirus genes involved in late and very late gene expression: ie-1, ie-n, and lef-2. J Virol. 1993 Apr;67(4):2149–2158. doi: 10.1128/jvi.67.4.2149-2158.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pearson M. N., Russell R. L., Rohrmann G. F., Beaudreau G. S. p39, a major baculovirus structural protein: immunocytochemical characterization and genetic location. Virology. 1988 Dec;167(2):407–413. [PubMed] [Google Scholar]
  25. Possee R. D., Sun T. P., Howard S. C., Ayres M. D., Hill-Perkins M., Gearing K. L. Nucleotide sequence of the Autographa californica nuclear polyhedrosis 9.4 kbp EcoRI-I and -R (polyhedrin gene) region. Virology. 1991 Nov;185(1):229–241. doi: 10.1016/0042-6822(91)90770-c. [DOI] [PubMed] [Google Scholar]
  26. Rohrmann G. F. Baculovirus structural proteins. J Gen Virol. 1992 Apr;73(Pt 4):749–761. doi: 10.1099/0022-1317-73-4-749. [DOI] [PubMed] [Google Scholar]
  27. Russell R. L., Rohrmann G. F. A baculovirus polyhedron envelope protein: immunogold localization in infected cells and mature polyhedra. Virology. 1990 Jan;174(1):177–184. doi: 10.1016/0042-6822(90)90066-z. [DOI] [PubMed] [Google Scholar]
  28. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Theilmann D. A., Chantler J. K., Stweart S., Flipsen H. T., Vlak J. M., Crook N. E. Characterization of a highly conserved baculovirus structural protein that is specific for occlusion-derived virions. Virology. 1996 Apr 1;218(1):148–158. doi: 10.1006/viro.1996.0175. [DOI] [PubMed] [Google Scholar]
  30. Thiem S. M., Miller L. K. Identification, sequence, and transcriptional mapping of the major capsid protein gene of the baculovirus Autographa californica nuclear polyhedrosis virus. J Virol. 1989 May;63(5):2008–2018. doi: 10.1128/jvi.63.5.2008-2018.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Vaughn J. L., Goodwin R. H., Tompkins G. J., McCawley P. The establishment of two cell lines from the insect Spodoptera frugiperda (Lepidoptera; Noctuidae). In Vitro. 1977 Apr;13(4):213–217. doi: 10.1007/BF02615077. [DOI] [PubMed] [Google Scholar]
  32. Vialard J. E., Richardson C. D. The 1,629-nucleotide open reading frame located downstream of the Autographa californica nuclear polyhedrosis virus polyhedrin gene encodes a nucleocapsid-associated phosphoprotein. J Virol. 1993 Oct;67(10):5859–5866. doi: 10.1128/jvi.67.10.5859-5866.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wilson M. E., Mainprize T. H., Friesen P. D., Miller L. K. Location, transcription, and sequence of a baculovirus gene encoding a small arginine-rich polypeptide. J Virol. 1987 Mar;61(3):661–666. doi: 10.1128/jvi.61.3.661-666.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wolgamot G. M., Gross C. H., Russell R. L., Rohrmann G. F. Immunocytochemical characterization of p24, a baculovirus capsid-associated protein. J Gen Virol. 1993 Jan;74(Pt 1):103–107. doi: 10.1099/0022-1317-74-1-103. [DOI] [PubMed] [Google Scholar]

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