1887

Abstract

SUMMARY

Adenovirus H5 (Ad5) DNA–protein complexes were extracted with ammonium sulphate (0.2 ) from virus-infected HeLa cell nuclei at 18 h after infection. Analysis of the material by centrifugation through discontinuous sucrose gradients in heavy water revealed the existence of several populations of molecules which were identified, in order of increasing buoyant density, as mature DNA–protein complexes, replication complexes, assembly intermediates and virions. When observed under the electron microscope, some of the assembly intermediates showed a capsid with a tail of entirely double-stranded (ds) DNA, or of dsDNA continued by a portion of single-stranded (ss) DNA thickened by a coat of E-72 K DNA binding protein. Singly or doubly-forked Ad5 replicating DNA molecules partially packaged in virus capsids were also observed. It is suggested that these molecules could be assembly intermediates, i.e. one of the first steps of assembly corresponding to virus DNA entering pre-formed capsids or their precursors. The fact that replication was still going on at one end of many of the DNA molecules in the intermediates, while encapsidation was taking place at the other, raises the possibility of a coupled DNA replication-packaging process in the formation of adenovirions.

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1980-09-01
2021-10-20
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References

  1. Anderson C. W., Baum P. R., Gesteland R. F. 1973; Processing of adenovirus 2-induced proteins. Journal of Virology 12:241–252
    [Google Scholar]
  2. Baltimore D., Huang A. S. 1968; Isopycnic separation of subcellular components from poliovirus infected and normal HeLa cells. Science 162:572–574
    [Google Scholar]
  3. Brison O., Kedinger C., Wilhelm J. 1977; Enzymatic properties of viral replication complexes isolated from adenovirus type 2-infected HeLa cell nuclei. Journal of Virology 24:423–435
    [Google Scholar]
  4. Brown D. T., Westphal M., Burlingham B. T., Winterhoff U., Doerfler W. 1975; Structure and composition of the adenovirus type 2 core. Journal of Virology 16:366–387
    [Google Scholar]
  5. Corden J., Engelking H. M., Pearson G. D. 1976; Chromatin-like organization of the adenovirus chromosome. Proceedings of the National Academy of Sciences of the United States of America 73:401–404
    [Google Scholar]
  6. Daniell E. 1976; Genome structure of incomplete particles of adenovirus. Journal of Virology 19:685–708
    [Google Scholar]
  7. D’halluin J. C., Milleville M., Boulanger P. A., Martin G. R. 1978a; Temperature-sensitive mutant of adenovirus type 2 blocked in virion assembly: accumulation of light intermediate particles. Journal of Virology 26:344–356
    [Google Scholar]
  8. D’halluin J. C., Martin G. R., Torpier G., Boulanger P. A. 1978b; Adenovirus type 2 assembly analysed by reversible cross-linking of labile intermediates. Journal of Virology 26:357–363
    [Google Scholar]
  9. Dubochet J., Ducommun M., Zollinger M., Kellenberger E. 1971; A new preparation method for dark field electron microscopy of biomacromolecules. Journal of Ultrastructural Research 35:147–167
    [Google Scholar]
  10. Edvarsson B., Everjtt E., Jornwall H., Prage L., Philipson L. 1976; Intermediates in adenovirus assembly. Journal of Virology 19:533–547
    [Google Scholar]
  11. Edvarrson B., Ustacelebi S., Williams J., Philipson L. 1978; Assembly intermediates among adenovirus type 5 temperature-sensitive mutants. Journal of Virology 25:641–651
    [Google Scholar]
  12. Ellens D. J., Sussenbach J. S., Jansz H. S. 1974; Studies on the mechanism of replication of adenovirus DNA. III. Electron microscopy of replicating DNA. Virology 61:427–442
    [Google Scholar]
  13. Ensinger M. J., Ginsberg H. S. 1972; Selection and characterization of temperature-sensitive mutants of type 5 adenovirus. Journal of Virology io:328–339
    [Google Scholar]
  14. Everitt E., Sundquist B., Pettersson U., Philipson L. 1973; Structural proteins of adenovirus. X. Isolation and topography of low molecular weight antigens from the virion of adenovirus type 2. Virology 52:130–147
    [Google Scholar]
  15. Girard M., Bouche J. P., Marty L., Revet B., Berthelot N. 1977; Circular adenovirus DNA-protein complexes from infected HeLa cell nuclei. Virology 83:34–55
    [Google Scholar]
  16. Ishibashi M., Maizel I. V. Jun 1974; The polypeptides of adenovirus. V. Young virions, structural intermediate between top components and aged virions. Virology 58:345–361
    [Google Scholar]
  17. Ishibashi M., Chin W. W., Maizel J. V. Jun 1977; The polypeptides of adenovirus. IX-Partial purification of early proteins and observations on late proteins of type 5 adenovirus. Virology 83:1–15
    [Google Scholar]
  18. Kedinger C., Brison O., Perrin F., Wilhelm J. 1978; Structural analysis of viral replicative intermediates isolated from adenovirus type 2-infected HeLa cell nuclei. Journal of Virology 26:364–379
    [Google Scholar]
  19. Kit G., Daniell E. 1978; Adenovirus core protein synthesis in the absence of viral DNA synthesis late in infection. Journal of Virology 27:74–80
    [Google Scholar]
  20. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature London 222:680–685
    [Google Scholar]
  21. Laskey R. A., Mills A. D. 1975; Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. European Journal of Biochemistry 56:335–341
    [Google Scholar]
  22. Laver W. G. 1970; Isolation of an arginine-rich protein from particles of adenovirus type 2. Virology 41:488–500
    [Google Scholar]
  23. Lechner R. L., Kelly T. J. 1977; The structure of replicating adenovirus DNA molecules. Cell 12:1007–1020
    [Google Scholar]
  24. Levine A. J., Van Der Vliet P. C., Rosenwirth B., Rabek J., Frenkel G., Ensinger M. S. 1974; Adenovirus infected, cell-specific, DNA-binding proteins. Cold Spring Harbor Symposia on Quantitative Biology 39:559–566
    [Google Scholar]
  25. Levine A. J., Van Der Vliet P. C., Rosenwirth B., Anderson C., Rabek J., Levinson A., Anderson S. 1976; Characterization of an adenovirus early protein required for virus DNA replication: a singlestranded DNA-binding protein. Molecular and Cellular Biochemistry 11:79–95
    [Google Scholar]
  26. Matsuguchi M., Puvion-DuTilleul F., Moyne G. 1979; Late transcription and simultaneous replication of simian adenovirus 7 DNA as revealed by spreading lytically infected cell cultures. Journal of General Virology 42:443–456
    [Google Scholar]
  27. Philipson P., Lindberg U. 1974; Reproduction of adenoviruses. In Comprehensive Virology vol 3 pp 143–227 Edited by Fraenkel-Conrat H., Wagner R. R. New York and London: Plenum Press;
    [Google Scholar]
  28. Prage L., Pettersson U. 1971; Structural proteins of adenoviruses. VII. Purification and properties of an arginine-rich core protein from adenovirus type 2 and type 3. Virology 45:364–373
    [Google Scholar]
  29. Rekosh D. M., Russell W. C., Bellett A. J. D., Robinson A. J. 1977; Identification of a protein linked to the ends of adenovirus DNA. Cell xi:283–295
    [Google Scholar]
  30. Rickwood D., Birnie G.D. 1975; Metrizamide, a new density gradient medium. FEBS Letters 50:102–109
    [Google Scholar]
  31. Tattersall P., Ward D. C. 1976; Rolling hairpin model for replication of parvoviruses and linear chromosomal DNA. Nature, London 263:106–109
    [Google Scholar]
  32. Van Der Vliet P. C., Levine A. J. 1973; DNA binding proteins specific for cells infected by adenovirus. Nature New Biology 146:170–174
    [Google Scholar]
  33. Van Der Vliet P. C., Levine A. J., Ensinger M. J., Ginsberg H. S. 1975; Thermolabile DNA-binding proteins from cells infected with a temperature sensitive mutant of adenovirus defective in viral DNA synthesis. Journal of Virology 15:348–354
    [Google Scholar]
  34. Van Der Vliet P. C., Zandberg J., Jansz H. S. 1977; Evidence for a function of the adenovirus DNA-binding protein in initiation of DNA synthesis as well as in elongation of nascent DNA chains. Virology 80:98–110
    [Google Scholar]
  35. Weber J. 1976; Genetic analysis of adenovirus type 2. III. Temperature sensitivity of processing of viral proteins. Journal of Virology 17:462–471
    [Google Scholar]
  36. Wilhelm J., Brison O., Kedinger C., Chambon P. 1976; Characterization of adenovirus type 2 transcriptional complexes isolated from infected HeLa cell nuclei. Journal of Virology 19:61–81
    [Google Scholar]
  37. Windberg G., Wadell G. 1977; Structural polypeptide of adenovirus type 16 incomplete particles. Journal of Virology 22:389–401
    [Google Scholar]
  38. Winnacker E. L. 1978; Adenovirus DNA: structure and function of a novel replicon. Cell 14:761–773
    [Google Scholar]
  39. Yamagishi H., Okamoto M. 1978; Visualization of the intracellular development of bacteriophage A, with special reference to DNA packaging. Proceedings of the National Academy of Sciences of the United States of America 75:3206–3210
    [Google Scholar]
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