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. 1978 May;26(2):344–356. doi: 10.1128/jvi.26.2.344-356.1978

Temperature-sensitive mutant of adenovirus type 2 blocked in virion assembly: accumulation of light intermediate particles.

J C D'Halluin, M Milleville, P A Boulanger, G R Martin
PMCID: PMC354073  PMID: 660717

Abstract

A temperature-sensitive mutant of human adenovirus type 2, ts112, was isolated and characterized, ts112 was blocked in a late function required for virus maturation. At restrictive temperature, it accumulated light precursor particles that were able to mature into infectious virions upon temperature shift-down. Use of a mild extraction procedure and a reversible fixation by a cleavable diimido ester permitted the isolation and analysis of these labile intermediates in the adenovirus assembly. These accumulated particles had a sedimentation coefficient of about 600S and a buoyant density of 1.315 g/cm3 in CsCl. They contained a DNA fragment of 7--11S and two nonvirion proteins having molecular weights of 50,000 (50K) and 39.000 (39K), respectively. They resembled in composition and morphology the light intermediate particles found in wild-type adenovirus 2, which were identified as precursors of heavy intermediates, preceding the young virions. The ts112 lesion was apparently located at the exit of either the 50K and/or 39K proteins and at the entry of viral DNA.

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Selected References

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  1. Anderson C. W., Baum P. R., Gesteland R. F. Processing of adenovirus 2-induced proteins. J Virol. 1973 Aug;12(2):241–252. doi: 10.1128/jvi.12.2.241-252.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Botstein D., Waddell C. H., King J. Mechanism of head assembly and DNA encapsulation in Salmonella phage p22. I. Genes, proteins, structures and DNA maturation. J Mol Biol. 1973 Nov 15;80(4):669–695. doi: 10.1016/0022-2836(73)90204-0. [DOI] [PubMed] [Google Scholar]
  3. Boulanger P. A., Puvion F. Adenovirus assembly. Cross-linking of adenovirus type-2 hexons in vitro. Eur J Biochem. 1974 Apr 16;43(3):465–470. doi: 10.1111/j.1432-1033.1974.tb03433.x. [DOI] [PubMed] [Google Scholar]
  4. Boulanger P. A., Puvion F. Large-scale preparation of soluble adenovirus hexon, penton and fiber antigens in highly purified form. Eur J Biochem. 1973 Nov 1;39(1):37–42. doi: 10.1111/j.1432-1033.1973.tb03100.x. [DOI] [PubMed] [Google Scholar]
  5. Burlingham B. T., Brown D. T., Doerfler W. Incomplete particles of adenovirus. I. Characteristics of the DNA associated with incomplete adenovirions of types 2 and 12. Virology. 1974 Aug;60(2):419–430. doi: 10.1016/0042-6822(74)90336-5. [DOI] [PubMed] [Google Scholar]
  6. Casjens S., King J. Virus assembly. Annu Rev Biochem. 1975;44:555–611. doi: 10.1146/annurev.bi.44.070175.003011. [DOI] [PubMed] [Google Scholar]
  7. D'Halluin J. C., Martin G. R., Torpier G., Boulanger P. A. Adenovirus type 2 assembly analyzed by reversible cross-linking of labile intermediates. J Virol. 1978 May;26(2):357–363. doi: 10.1128/jvi.26.2.357-363.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Daniell E. Genome structure of incomplete particles of adenovirus. J Virol. 1976 Aug;19(2):685–708. doi: 10.1128/jvi.19.2.685-708.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Doerfler W. Nonproductive infection of baby hamster kidney cells (BHK21) with adenovirus type 12. Virology. 1969 Aug;38(4):587–606. doi: 10.1016/0042-6822(69)90179-2. [DOI] [PubMed] [Google Scholar]
  10. Edvardsson B., Everitt E., Jörnvall H., Prage L., Philipson L. Intermediates in adenovirus assembly. J Virol. 1976 Aug;19(2):533–547. doi: 10.1128/jvi.19.2.533-547.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Eiserling F. A., Dickson R. C. Assembly of viruses. Annu Rev Biochem. 1972;41:467–502. doi: 10.1146/annurev.bi.41.070172.002343. [DOI] [PubMed] [Google Scholar]
  12. Everitt E., Meador S. A., Levine A. S. Synthesis and processing of the precursor to the major core protein of adenovirus type 2. J Virol. 1977 Jan;21(1):199–214. doi: 10.1128/jvi.21.1.199-214.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ishibashi M., Maizel J. V., Jr The polypeptides of adenovirus. V. Young virions, structural intermediate between top components and aged virions. Virology. 1974 Feb;57(2):409–424. doi: 10.1016/0042-6822(74)90181-0. [DOI] [PubMed] [Google Scholar]
  14. Khittoo G., Weber J. Genetic analysis of adenovirus type 2. VI. A temperature-sensitive mutant defective for DNA encapsidation. Virology. 1977 Aug;81(1):126–137. doi: 10.1016/0042-6822(77)90064-2. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Oberg B., Saborio J., Persson T., Everitt E., Philipson L. Identification of the in vitro translation products of adenovirus mRNA by immunoprecipitation. J Virol. 1975 Jan;15(1):199–207. doi: 10.1128/jvi.15.1.199-207.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Rosenwirth B., Tjia S., Westphal M., Doerfler W. Incomplete particles of adenovirus. II. Kinetics of formation and polypeptide composition of adenovirus type 2. Virology. 1974 Aug;60(2):431–437. doi: 10.1016/0042-6822(74)90337-7. [DOI] [PubMed] [Google Scholar]
  18. Saborio J. L., Oberg B. In vivo and in vitro synthesis of adenovirus type 2 early proteins. J Virol. 1976 Mar;17(3):865–875. doi: 10.1128/jvi.17.3.865-875.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sundquist B., Everitt E., Philipson L., Hoglund S. Assembly of adenoviruses. J Virol. 1973 Mar;11(3):449–459. doi: 10.1128/jvi.11.3.449-459.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tjia S., Fanning E., Schick J., Doerfler W. Incomplete particles of adenovirus type 2. III. Viral and cellular DNA sequences in incomplete particles. Virology. 1977 Jan;76(1):365–379. doi: 10.1016/0042-6822(77)90309-9. [DOI] [PubMed] [Google Scholar]
  21. Wallace R. D., Kates J. State of adenovirus 2 deoxyribonucleic acid in the nucleus and its mode of transcription: studies with isolated viral deoxyribonucleic acid-protein complexes and isolated nuclei. J Virol. 1972 Apr;9(4):627–635. doi: 10.1128/jvi.9.4.627-635.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Warocquier R., Boulanger P. A. Adenovirus protein maturation at 42 degrees C. J Gen Virol. 1976 Feb;30(2):233–242. doi: 10.1099/0022-1317-30-2-233. [DOI] [PubMed] [Google Scholar]
  23. Weber J. Genetic analysis of adenovirus type 2 III. Temperature sensitivity of processing viral proteins. J Virol. 1976 Feb;17(2):462–471. doi: 10.1128/jvi.17.2.462-471.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Winberg G., Wadell G. Structural polypeptides of adenovirus type 16 incomplete particles. J Virol. 1977 May;22(2):389–401. doi: 10.1128/jvi.22.2.389-401.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Winters W. D., Russell W. C. Studies on the assembly of adenovirus in vitro. J Gen Virol. 1971 Feb;10(2):181–194. doi: 10.1099/0022-1317-10-2-181. [DOI] [PubMed] [Google Scholar]

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