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. 1992 Feb;66(2):723–731. doi: 10.1128/jvi.66.2.723-731.1992

cis and trans requirements for the selective packaging of adenovirus type 5 DNA.

M Gräble 1, P Hearing 1
PMCID: PMC240771  PMID: 1731109

Abstract

Polar packaging of adenovirus DNA into virions is dependent on the presence of cis-acting sequences at the left end of the viral genome. Our previous analyses demonstrated that the adenovirus type 5 (Ad5) packaging domain (nucleotides 194 to 358) is composed of at least five elements that are functionally redundant. A repeated sequence, termed the A repeat, was associated with packaging function. Here we report a more detailed analysis of the requirements for the selective packaging of Ad5 DNA. By introducing site-directed point mutations into specific A repeat sequences, we demonstrate that the A repeats represent cis-acting functional components of the packaging signal. Additional elements, located outside the originally defined packaging domain boundaries and that resemble the A repeat consensus sequence, also are capable of promoting the packaging of viral DNA. The cis-acting components of the packaging signal appear to be subject to certain spatial constraints for function, possibly reflecting a necessity for the coordinate binding of packaging proteins to these sites. In agreement with this idea, we present evidence that the interaction of a limiting trans-acting factor(s) with the packaging domain in vivo is required for efficient encapsidation of the Ad5 genome.

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Selected References

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  1. Bruder J. T., Hearing P. Nuclear factor EF-1A binds to the adenovirus E1A core enhancer element and to other transcriptional control regions. Mol Cell Biol. 1989 Nov;9(11):5143–5153. doi: 10.1128/mcb.9.11.5143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chee-Sheung C. C., Ginsberg H. S. Characterization of a temperature-sensitive fiber mutant of type 5 adenovirus and effect of the mutation on virion assembly. J Virol. 1982 Jun;42(3):932–950. doi: 10.1128/jvi.42.3.932-950.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. 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]
  5. D'Halluin J. C., Milleville M., Boulanger P. Effects of novobiocin on adenovirus DNA synthesis and encapsidation. Nucleic Acids Res. 1980 Apr 11;8(7):1625–1641. doi: 10.1093/nar/8.7.1625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. D'Halluin J. C., Milleville M., Martin G. R., Boulanger P. Morphogenesis of human adenovirus type 2 studied with fiber- and fiber and penton base-defective temperature-sensitive mutants. J Virol. 1980 Jan;33(1):88–99. doi: 10.1128/jvi.33.1.88-99.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. DeLucia A. L., Deb S., Partin K., Tegtmeyer P. Functional interactions of the simian virus 40 core origin of replication with flanking regulatory sequences. J Virol. 1986 Jan;57(1):138–144. doi: 10.1128/jvi.57.1.138-144.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Earnshaw W. C., Casjens S. R. DNA packaging by the double-stranded DNA bacteriophages. Cell. 1980 Sep;21(2):319–331. doi: 10.1016/0092-8674(80)90468-7. [DOI] [PubMed] [Google Scholar]
  10. Eckdahl T. T., Anderson J. N. Bent DNA is a conserved structure in an adenovirus control region. Nucleic Acids Res. 1988 Mar 25;16(5):2346–2346. doi: 10.1093/nar/16.5.2346. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Edvardsson B., Ustacelebi S., Williams J., Philipson L. Assembly intermediates among adenovirus type 5 temperature-sensitive mutants. J Virol. 1978 Feb;25(2):641–651. doi: 10.1128/jvi.25.2.641-651.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  14. Gluzman Y. SV40-transformed simian cells support the replication of early SV40 mutants. Cell. 1981 Jan;23(1):175–182. doi: 10.1016/0092-8674(81)90282-8. [DOI] [PubMed] [Google Scholar]
  15. Graham F. L., Smiley J., Russell W. C., Nairn R. Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol. 1977 Jul;36(1):59–74. doi: 10.1099/0022-1317-36-1-59. [DOI] [PubMed] [Google Scholar]
  16. Gräble M., Hearing P. Adenovirus type 5 packaging domain is composed of a repeated element that is functionally redundant. J Virol. 1990 May;64(5):2047–2056. doi: 10.1128/jvi.64.5.2047-2056.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hammarskjöld M. L., Winberg G. Encapsidation of adenovirus 16 DNA is directed by a small DNA sequence at the left end of the genome. Cell. 1980 Jul;20(3):787–795. doi: 10.1016/0092-8674(80)90325-6. [DOI] [PubMed] [Google Scholar]
  18. Hasson T. B., Soloway P. D., Ornelles D. A., Doerfler W., Shenk T. Adenovirus L1 52- and 55-kilodalton proteins are required for assembly of virions. J Virol. 1989 Sep;63(9):3612–3621. doi: 10.1128/jvi.63.9.3612-3621.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hearing P., Samulski R. J., Wishart W. L., Shenk T. Identification of a repeated sequence element required for efficient encapsidation of the adenovirus type 5 chromosome. J Virol. 1987 Aug;61(8):2555–2558. doi: 10.1128/jvi.61.8.2555-2558.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hearing P., Shenk T. The adenovirus type 5 E1A enhancer contains two functionally distinct domains: one is specific for E1A and the other modulates all early units in cis. Cell. 1986 Apr 25;45(2):229–236. doi: 10.1016/0092-8674(86)90387-9. [DOI] [PubMed] [Google Scholar]
  21. Hearing P., Shenk T. The adenovirus type 5 E1A transcriptional control region contains a duplicated enhancer element. Cell. 1983 Jul;33(3):695–703. doi: 10.1016/0092-8674(83)90012-0. [DOI] [PubMed] [Google Scholar]
  22. Jones N., Shenk T. Isolation of adenovirus type 5 host range deletion mutants defective for transformation of rat embryo cells. Cell. 1979 Jul;17(3):683–689. doi: 10.1016/0092-8674(79)90275-7. [DOI] [PubMed] [Google Scholar]
  23. Kosturko L. D., Sharnick S. V., Tibbetts C. Polar encapsidation of adenovirus DNA: cloning and DNA sequence of the left end of adenovirus type 3. J Virol. 1982 Sep;43(3):1132–1137. doi: 10.1128/jvi.43.3.1132-1137.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kovesdi I., Reichel R., Nevins J. R. Role of an adenovirus E2 promoter binding factor in E1A-mediated coordinate gene control. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2180–2184. doi: 10.1073/pnas.84.8.2180. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kunkel T. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci U S A. 1985 Jan;82(2):488–492. doi: 10.1073/pnas.82.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Ohyama T., Hashimoto S. Upstream half of adenovirus type 2 enhancer adopts a curved DNA conformation. Nucleic Acids Res. 1989 May 25;17(10):3845–3853. doi: 10.1093/nar/17.10.3845. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Robinson C. C., Tibbetts C. Polar encapsidation of adenovirus DNA: evolutionary variants reveal dispensable sequences near the left ends of Ad3 genomes. Virology. 1984 Sep;137(2):276–286. doi: 10.1016/0042-6822(84)90219-8. [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. Stow N. D. Cloning of a DNA fragment from the left-hand terminus of the adenovirus type 2 genome and its use in site-directed mutagenesis. J Virol. 1981 Jan;37(1):171–180. doi: 10.1128/jvi.37.1.171-180.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Tibbetts C. Viral DNA sequences from incomplete particles of human adenovirus type 7. Cell. 1977 Sep;12(1):243–249. doi: 10.1016/0092-8674(77)90202-1. [DOI] [PubMed] [Google Scholar]
  32. Wigler M., Silverstein S., Lee L. S., Pellicer A., Cheng Y. c., Axel R. Transfer of purified herpes virus thymidine kinase gene to cultured mouse cells. Cell. 1977 May;11(1):223–232. doi: 10.1016/0092-8674(77)90333-6. [DOI] [PubMed] [Google Scholar]

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