Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1978 May;26(2):357–363. doi: 10.1128/jvi.26.2.357-363.1978

Adenovirus type 2 assembly analyzed by reversible cross-linking of labile intermediates.

J C D'Halluin, G R Martin, G Torpier, P A Boulanger
PMCID: PMC354074  PMID: 660718

Abstract

Dimethyl-4,4'-dithiobisbutyrimidate dihydrochloride was used as a cleavable cross-linking reagent to maintain the structure of labile intermediates in adenovirus type 2 assembly. Analysis on sucrose gradients of nuclear adenovirus particles revealed two size classes, with sedimentation rates of 750 and 600S. After reversible fixation with diimido ester, the different classes were further separated on CsCl gradients and characterized with regard to their buoyant density, DNA content, and polypeptide composition. The 750S particles banded at 1.345 g/cm3 in CsCl, contained a DNA with a sedimentation coefficient of 34S in alkaline sucrose gradients, and had a polypeptide composition similar to that of young virions. The 600S population consisted of two types of particles with buoyant densities of 1.315 and 1.37 g/cm3. The 1.315-g/cm3 particles contained a DNA fragment of 7--11S and lacked the core proteins V and VII. In their place were found precursors P VI and P VIII and two nonvirion proteins with molecular weights of 50,000 (50K) and 39,000 (39K). 34S DNA was present in the 1.37-g/cm3 particles, which also lacked core proteins V and VII, as well as the 50K and 39K. Pulse-chase labeling kinetics suggested that the 1.315-g/cm3 particles were anterior to the 1.37-g/cm3 particles, themselves preceding the 1.345-g/cm3 young virions, and that the release of both 50K and 39K, possible scaffolding proteins, was required for entry of viral DNA.

Full text

PDF
358

Images in this article

360Image
on p.360
362Image
on p.362

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. 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]
  2. D'Halluin J. C., Milleville M., Boulanger P. A., Martin G. R. Temperature-sensitive mutant of adenovirus type 2 blocked in virion assembly: accumulation of light intermediate particles. J Virol. 1978 May;26(2):344–356. doi: 10.1128/jvi.26.2.344-356.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. 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]
  5. 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]
  6. GREEN M., PINA M. Biochemical studies on adenovirus multiplication. IV. Isolation, purification, and chemical analysis of adenovirus. Virology. 1963 May;20:199–207. doi: 10.1016/0042-6822(63)90157-0. [DOI] [PubMed] [Google Scholar]
  7. Harpst J. A., Ennever J. F., Russell W. C. Physical properties of nucleoprotein cores from adenovirus type 5. Nucleic Acids Res. 1977 Feb;4(2):477–490. doi: 10.1093/nar/4.2.477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hennache B., Boulanger P. Biochemical study of KB-cell receptor for adenovirus. Biochem J. 1977 Aug 15;166(2):237–247. doi: 10.1042/bj1660237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. 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]
  11. Philipson L., Lonberg-Holm K., Pettersson U. Virus-receptor interaction in an adenovirus system. J Virol. 1968 Oct;2(10):1064–1075. doi: 10.1128/jvi.2.10.1064-1075.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Traut R. R., Bollen A., Sun T. T., Hershey J. W., Sundberg J., Pierce L. R. Methyl 4-mercaptobutyrimidate as a cleavable cross-linking reagent and its application to the Escherichia coli 30S ribosome. Biochemistry. 1973 Aug 14;12(17):3266–3273. doi: 10.1021/bi00741a019. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. van der Vliet P. C., Levine A. J. DNA-binding proteins specific for cells infected by adenovirus. Nat New Biol. 1973 Dec 12;246(154):170–174. doi: 10.1038/newbio246170a0. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES