1887

Abstract

Adenovirus (AdV) is thought to follow a sequential assembly pathway similar to that observed in dsDNA bacteriophages and herpesviruses. First, empty capsids are assembled, and then the genome is packaged through a ring-like structure, referred to as a portal, located at a unique vertex. In human AdV serotype 5 (HAdV5), the IVa2 protein initiates specific recognition of viral genome by associating with the viral packaging domain located between nucleotides 220 and 400 of the genome. IVa2 is located at a unique vertex on mature capsids and plays an essential role during genome packaging, most likely by acting as a DNA packaging ATPase. In this study, we demonstrated interactions among IVa2, 33K and DNA-binding protein (DBP) in virus-infected cells by cross-linking of HAdV5-infected cells followed by Western blot, and co-immunoprecipitation of IVa2, 33K and DBP from nuclear extracts of HAdV5-infected cells. Confocal microscopy demonstrated co-localization of IVa2, 33K and DBP in virus-infected cells and also in cells transfected with IVa2, 33K and DBP genes. Immunogold electron microscopy of purified HAdV5 showed the presence of IVa2, 33K or DBP at a single site on the virus particles. Our results provide indirect evidence that IVa2, 33K and DBP may form a complex at a unique vertex on viral capsids and cooperate in genome packaging.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.049346-0
2013-06-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jgv/94/6/1325.html?itemId=/content/journal/jgv/10.1099/vir.0.049346-0&mimeType=html&fmt=ahah

References

  1. Ali H., LeRoy G., Bridge G., Flint S. J. 2007; The adenovirus L4 33-kilodalton protein binds to intragenic sequences of the major late promoter required for late phase-specific stimulation of transcription. J Virol 81:1327–1338 [View Article][PubMed]
    [Google Scholar]
  2. Bangari D. S., Mittal S. K. 2004; Porcine adenoviral vectors evade preexisting humoral immunity to adenoviruses and efficiently infect both human and murine cells in culture. Virus Res 105:127–136 [View Article][PubMed]
    [Google Scholar]
  3. Bosher J., Dawson A., Hay R. T. 1992; Nuclear factor I is specifically targeted to discrete subnuclear sites in adenovirus type 2-infected cells. J Virol 66:3140–3150[PubMed]
    [Google Scholar]
  4. Christensen J. B., Byrd S. A., Walker A. K., Strahler J. R., Andrews P. C., Imperiale M. J. 2008; Presence of the adenovirus IVa2 protein at a single vertex of the mature virion. J Virol 82:9086–9093 [View Article][PubMed]
    [Google Scholar]
  5. Daniell E. 1976; Genome structure of incomplete particles of adenovirus. J Virol 19:685–708[PubMed]
    [Google Scholar]
  6. Dittmer A., Drach J. C., Townsend L. B., Fischer A., Bogner E. 2005; Interaction of the putative human cytomegalovirus portal protein pUL104 with the large terminase subunit pUL56 and its inhibition by benzimidazole-D-ribonucleosides. J Virol 79:14660–14667 [View Article][PubMed]
    [Google Scholar]
  7. Edvardsson B., Everitt E., Jörnvall H., Prage L., Philipson L. 1976; Intermediates in adenovirus assembly. J Virol 19:533–547[PubMed]
    [Google Scholar]
  8. Ewing S. G., Byrd S. A., Christensen J. B., Tyler R. E., Imperiale M. J. 2007; Ternary complex formation on the adenovirus packaging sequence by the IVa2 and L4 22-kilodalton proteins. J Virol 81:12450–12457 [View Article][PubMed]
    [Google Scholar]
  9. Fu C. Y., Prevelige P. E. Jr 2009; In vitro incorporation of the phage Phi29 connector complex. Virology 394:149–153 [View Article][PubMed]
    [Google Scholar]
  10. Giard D. J., Aaronson S. A., Todaro G. J., Arnstein P., Kersey J. H., Dosik H., Parks W. P. 1973; In vitro cultivation of human tumors: establishment of cell lines derived from a series of solid tumors. J Natl Cancer Inst 51:1417–1423[PubMed]
    [Google Scholar]
  11. Gonzalez R. A., Flint S. J. 2002; Effects of mutations in the adenoviral E1B 55-kilodalton protein coding sequence on viral late mRNA metabolism. J Virol 76:4507–4519 [View Article][PubMed]
    [Google Scholar]
  12. Gräble M., Hearing P. 1990; Adenovirus type 5 packaging domain is composed of a repeated element that is functionally redundant. J Virol 64:2047–2056[PubMed]
    [Google Scholar]
  13. Graham F. L., Prevec L. 1991; Manipulation of Adenovirus Vectors. In Gene Transfer and Expression Protocols, 7th edn. pp. 109–128 Edited by Murray E. J. New York, NY: Humana Press; [View Article]
    [Google Scholar]
  14. Graham F. L., Smiley J., Russell W. C., Nairn R. 1977; Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol 36:59–72 [View Article][PubMed]
    [Google Scholar]
  15. Gustin K. E., Imperiale M. J. 1998; Encapsidation of viral DNA requires the adenovirus L1 52/55-kilodalton protein. J Virol 72:7860–7870[PubMed]
    [Google Scholar]
  16. Gustin K. E., Lutz P., Imperiale M. J. 1996; Interaction of the adenovirus L1 52/55-kilodalton protein with the IVa2 gene product during infection. J Virol 70:6463–6467[PubMed]
    [Google Scholar]
  17. Harada J. N., Shevchenko A., Shevchenko A., Pallas D. C., Berk A. J. 2002; Analysis of the adenovirus E1B-55K-anchored proteome reveals its link to ubiquitination machinery. J Virol 76:9194–9206 [View Article][PubMed]
    [Google Scholar]
  18. Hearing P., Samulski R. J., Wishart W. L., Shenk T. 1987; Identification of a repeated sequence element required for efficient encapsidation of the adenovirus type 5 chromosome. J Virol 61:2555–2558[PubMed]
    [Google Scholar]
  19. Horwitz M. S. 2001; Adenoviruses. In Fields Virology, 4th edn. pp. 2301–2326 Edited by Fields B. N., Knipe D. M., Howley P. M. Philadelphia, PA, USA: Lippincott Williams and Wilkins;
    [Google Scholar]
  20. Ishibashi M., Maizel J. V. Jr 1974; The polypeptides of adenovirus. V. Young virions, structural intermediate between top components and aged virions. Virology 57:409–424 [View Article][PubMed]
    [Google Scholar]
  21. Khittoo G., Weber J. M. 1981; The nature of the DNA associated with incomplete particles of adenovirus type 2. J Gen Virol 54:343–355 [View Article][PubMed]
    [Google Scholar]
  22. Kitchingman G. R. 1995; Mutations in the adenovirus-encoded single-stranded DNA binding protein that result in altered accumulation of early and late viral RNAs. Virology 212:91–101 [View Article][PubMed]
    [Google Scholar]
  23. Kosturko L. D., Sharnick S. V., Tibbetts C. 1982; Polar encapsidation of adenovirus DNA: cloning and DNA sequence of the left end of adenovirus type 3. J Virol 43:1132–1137[PubMed]
    [Google Scholar]
  24. Lutz P., Puvion-Dutilleul F., Lutz Y., Kedinger C. 1996; Nucleoplasmic and nucleolar distribution of the adenovirus IVa2 gene product. J Virol 70:3449–3460[PubMed]
    [Google Scholar]
  25. Newcomb W. W., Juhas R. M., Thomsen D. R., Homa F. L., Burch A. D., Weller S. K., Brown J. C. 2001; The UL6 gene product forms the portal for entry of DNA into the herpes simplex virus capsid. J Virol 75:10923–10932 [View Article][PubMed]
    [Google Scholar]
  26. Nicolas J. C., Sarnow P., Girard M., Levine A. J. 1983; Host range temperature-conditional mutants in the adenovirus DNA binding protein are defective in the assembly of infectious virus. Virology 126:228–239 [View Article][PubMed]
    [Google Scholar]
  27. Ostapchuk P., Hearing P. 2008; Adenovirus IVa2 protein binds ATP. J Virol 82:10290–10294 [View Article][PubMed]
    [Google Scholar]
  28. Ostapchuk P., Yang J., Auffarth E., Hearing P. 2005; Functional interaction of the adenovirus IVa2 protein with adenovirus type 5 packaging sequences. J Virol 79:2831–2838 [View Article][PubMed]
    [Google Scholar]
  29. Ostapchuk P., Anderson M. E., Chandrasekhar S., Hearing P. 2006; The L4 22-kilodalton protein plays a role in packaging of the adenovirus genome. J Virol 80:6973–6981 [View Article][PubMed]
    [Google Scholar]
  30. Ostapchuk P., Almond M., Hearing P. 2011; Characterization of Empty adenovirus particles assembled in the absence of a functional adenovirus IVa2 protein. J Virol 85:5524–5531 [View Article][PubMed]
    [Google Scholar]
  31. Pardo-Mateos A., Young C. S. 2004; A 40 kDa isoform of the type 5 adenovirus IVa2 protein is sufficient for virus viability. Virology 324:151–164 [View Article][PubMed]
    [Google Scholar]
  32. Parks R. J., Chen L., Anton M., Sankar U., Rudnicki M. A., Graham F. L. 1996; A helper-dependent adenovirus vector system: removal of helper virus by Cre-mediated excision of the viral packaging signal. Proc Natl Acad Sci U S A 93:13565–13570 [View Article][PubMed]
    [Google Scholar]
  33. Perez-Romero P., Tyler R. E., Abend J. R., Dus M., Imperiale M. J. 2005; Analysis of the interaction of the adenovirus L1 52/55-kilodalton and IVa2 proteins with the packaging sequence in vivo and in vitro. J Virol 79:2366–2374 [View Article][PubMed]
    [Google Scholar]
  34. Puvion-Dutilleul F., Puvion E. 1990; Replicating single-stranded adenovirus type 5 DNA molecules accumulate within well-delimited intranuclear areas of lytically infected HeLa cells. Eur J Cell Biol 52:379–388[PubMed]
    [Google Scholar]
  35. Rao V. B., Feiss M. 2008; The bacteriophage DNA packaging motor. Annu Rev Genet 42:647–681 [View Article][PubMed]
    [Google Scholar]
  36. Reich N. C., Sarnow P., Duprey E., Levine A. J. 1983; Monoclonal antibodies which recognize native and denatured forms of the adenovirus DNA-binding protein. Virology 128:480–484 [View Article][PubMed]
    [Google Scholar]
  37. Schmid S. I., Hearing P. 1997; Bipartite structure and functional independence of adenovirus type 5 packaging elements. J Virol 71:3375–3384[PubMed]
    [Google Scholar]
  38. Stuiver M. H., van der Vliet P. C. 1990; Adenovirus DNA-binding protein forms a multimeric protein complex with double-stranded DNA and enhances binding of nuclear factor I. J Virol 64:379–386[PubMed]
    [Google Scholar]
  39. Sun S., Rao V. B., Rossmann M. G. 2010; Genome packaging in viruses. Curr Opin Struct Biol 20:114–120 [View Article][PubMed]
    [Google Scholar]
  40. van Olphen A. L., Tikoo S. K., Mittal S. K. 2002; Characterization of bovine adenovirus type 3 E1 proteins and isolation of E1-expressing cell lines. Virology 295:108–118 [View Article][PubMed]
    [Google Scholar]
  41. Winter N., D’Halluin J. C. 1991; Regulation of the biosynthesis of subgroup C adenovirus protein IVa2. J Virol 65:5250–5259[PubMed]
    [Google Scholar]
  42. Zhang W., Imperiale M. J. 2003; Requirement of the adenovirus IVa2 protein for virus assembly. J Virol 77:3586–3594 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.049346-0
Loading
/content/journal/jgv/10.1099/vir.0.049346-0
Loading

Data & Media loading...

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