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. 1984 Nov;52(2):664–671. doi: 10.1128/jvi.52.2.664-671.1984

Deletion of the gene encoding the adenovirus 5 early region 1b 21,000-molecular-weight polypeptide leads to degradation of viral and host cell DNA.

S Pilder, J Logan, T Shenk
PMCID: PMC254571  PMID: 6492257

Abstract

The adenovirus 5 mutant H5dl337 lacks 146 base pairs within early region 1B. The deletion removes a portion of the region encoding the E1B 21,000-molecular-weight (21K) polypeptide, but does not disturb the E1B-55K/17K coding region. The virus is slightly defective for growth in cultured HeLa cells, in which its final yield is reduced ca. 10-fold compared with wild-type virus. The mutant displays a striking phenotype in HeLa cells. The onset of cytopathic effect is dramatically accelerated, and both host cell and viral DNAs are extensively degraded late after infection. This defect has been described previously for a variety of adenovirus mutants and has been termed a cytocidal (cyt) phenotype. H5dl337 serves to map this defect to the loss of E1B-21K polypeptide function. In addition to its defect in the productive growth cycle, H5dl337 is unable to transform rat cells at normal efficiency.

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Selected References

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  1. Aiello L., Guilfoyle R., Huebner K., Weinmann R. Adenovirus 5 DNA sequences present and RNA sequences transcribed in transformed human embryo kidney cells (HEK-Ad-5 or 293). Virology. 1979 Apr 30;94(2):460–469. doi: 10.1016/0042-6822(79)90476-8. [DOI] [PubMed] [Google Scholar]
  2. Anderson C. W., Schmitt R. C., Smart J. E., Lewis J. B. Early region 1B of adenovirus 2 encodes two coterminal proteins of 495 and 155 amino acid residues. J Virol. 1984 May;50(2):387–396. doi: 10.1128/jvi.50.2.387-396.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Babiss L. E., Ginsberg H. S. Adenovirus type 5 early region 1b gene product is required for efficient shutoff of host protein synthesis. J Virol. 1984 Apr;50(1):202–212. doi: 10.1128/jvi.50.1.202-212.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bernards R., Schrier P. I., Bos J. L., Van der Eb A. J. Role of adenovirus types 5 and 12 early region 1b tumor antigens in oncogenic transformation. Virology. 1983 May;127(1):45–53. doi: 10.1016/0042-6822(83)90369-0. [DOI] [PubMed] [Google Scholar]
  5. Blin N., Stafford D. W. A general method for isolation of high molecular weight DNA from eukaryotes. Nucleic Acids Res. 1976 Sep;3(9):2303–2308. doi: 10.1093/nar/3.9.2303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bos J. L., Polder L. J., Bernards R., Schrier P. I., van den Elsen P. J., van der Eb A. J., van Ormondt H. The 2.2 kb E1b mRNA of human Ad12 and Ad5 codes for two tumor antigens starting at different AUG triplets. Cell. 1981 Nov;27(1 Pt 2):121–131. doi: 10.1016/0092-8674(81)90366-4. [DOI] [PubMed] [Google Scholar]
  7. Chinnadurai G. Adenovirus 2 Ip+ locus codes for a 19 kd tumor antigen that plays an essential role in cell transformation. Cell. 1983 Jul;33(3):759–766. doi: 10.1016/0092-8674(83)90018-1. [DOI] [PubMed] [Google Scholar]
  8. Chinnadurai G., Chinnadurai S., Brusca J. Physical mapping of a large-plaque mutation of adenovirus type 2. J Virol. 1979 Nov;32(2):623–628. doi: 10.1128/jvi.32.2.623-628.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Esche H., Mathews M. B., Lewis J. B. Proteins and messenger RNAs of the transforming region of wild-type and mutant adenoviruses. J Mol Biol. 1980 Sep 25;142(3):399–417. doi: 10.1016/0022-2836(80)90279-x. [DOI] [PubMed] [Google Scholar]
  10. Ezoe H., Fatt R. B., Mak S. Degradation of intracellular DNA in KB cells infected with cyt mutants of human adenovirus type 12. J Virol. 1981 Oct;40(1):20–27. doi: 10.1128/jvi.40.1.20-27.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fisher P. B., Babiss L. E., Weinstein I. B., Ginsberg H. S. Analysis of type 5 adenovirus transformation with a cloned rat embryo cell line (CREF). Proc Natl Acad Sci U S A. 1982 Jun;79(11):3527–3531. doi: 10.1073/pnas.79.11.3527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Flint S. J., Gallimore P. H., Sharp P. A. Comparison of viral RNA sequences in adenovirus 2-transformed and lytically infected cells. J Mol Biol. 1975 Jul 25;96(1):47–68. doi: 10.1016/0022-2836(75)90181-3. [DOI] [PubMed] [Google Scholar]
  13. Freeman A. E., Black P. H., Vanderpool E. A., Henry P. H., Austin J. B., Huebner R. J. Transformation of primary rat embryo cells by adenovirus type 2. Proc Natl Acad Sci U S A. 1967 Sep;58(3):1205–1212. doi: 10.1073/pnas.58.3.1205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fukui Y., Saito I., Shiroki K., Shimojo H. Isolation of transformation-defective, replication-nondefective early region 1B mutants of adenovirus 12. J Virol. 1984 Jan;49(1):154–161. doi: 10.1128/jvi.49.1.154-161.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gallimore P. H. Viral DNA in transformed cells. II. A study of the sequences of adenovirus 2 DNA IN NINE LINES OF TRANSFORMED RAT CELLS USING SPECIFIC FRAGMENTS OF THE VIRAL GENOME;. J Mol Biol. 1974 Oct 15;89(1):49–72. doi: 10.1016/0022-2836(74)90162-4. [DOI] [PubMed] [Google Scholar]
  16. Gallimore P. H., Williams J. An examination of adenovirus type 5 mutants for their ability to induce group C adenovirus tumor-specific transplantation antigenicity in rats. Virology. 1982 Jul 15;120(1):146–156. doi: 10.1016/0042-6822(82)90013-7. [DOI] [PubMed] [Google Scholar]
  17. Gingeras T. R., Sciaky D., Gelinas R. E., Bing-Dong J., Yen C. E., Kelly M. M., Bullock P. A., Parsons B. L., O'Neill K. E., Roberts R. J. Nucleotide sequences from the adenovirus-2 genome. J Biol Chem. 1982 Nov 25;257(22):13475–13491. [PubMed] [Google Scholar]
  18. Graham F. L., Abrahams P. J., Mulder C., Heijneker H. L., Warnaar S. O., De Vries F. A., Fiers W., Van Der Eb A. J. Studies on in vitro transformation by DNA and DNA fragments of human adenoviruses and simian virus 40. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):637–650. doi: 10.1101/sqb.1974.039.01.077. [DOI] [PubMed] [Google Scholar]
  19. Graham F. L., Harrison T., Williams J. Defective transforming capacity of adenovirus type 5 host-range mutants. Virology. 1978 May 1;86(1):10–21. doi: 10.1016/0042-6822(78)90003-x. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Green M., Brackmann K. H., Cartas M. A., Matsuo T. Identification and purification of a protein encoded by the human adenovirus type 2 transforming region. J Virol. 1982 Apr;42(1):30–41. doi: 10.1128/jvi.42.1.30-41.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Green M., Brackmann K. H., Lucher L. A., Symington J. S., Kramer T. A. Human adenovirus 2 E1B-19K and E1B-53K tumor antigens: antipeptide antibodies targeted to the NH2 and COOH termini. J Virol. 1983 Dec;48(3):604–615. doi: 10.1128/jvi.48.3.604-615.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Harrison T., Graham F., Williams J. Host-range mutants of adenovirus type 5 defective for growth in HeLa cells. Virology. 1977 Mar;77(1):319–329. doi: 10.1016/0042-6822(77)90428-7. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  27. Ho Y. S., Galos R., Williams J. Isolation of type 5 adenovirus mutants with a cold-sensitive host range phenotype: genetic evidence of an adenovirus transformation maintenance function. Virology. 1982 Oct 15;122(1):109–124. doi: 10.1016/0042-6822(82)90381-6. [DOI] [PubMed] [Google Scholar]
  28. Jones N., Shenk T. An adenovirus type 5 early gene function regulates expression of other early viral genes. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3665–3669. doi: 10.1073/pnas.76.8.3665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Jones N., Shenk T. Isolation of deletion and substitution mutants of adenovirus type 5. Cell. 1978 Jan;13(1):181–188. doi: 10.1016/0092-8674(78)90148-4. [DOI] [PubMed] [Google Scholar]
  31. Katze M. G., Persson H., Philipson L. A novel mRNA and a low molecular weight polypeptide encoded in the transforming region of adenovirus DNA. EMBO J. 1982;1(7):783–789. doi: 10.1002/j.1460-2075.1982.tb01248.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Lai Fatt R. B., Mak S. Mapping of an adenovirus function involved in the inhibition of DNA degradation. J Virol. 1982 Jun;42(3):969–977. doi: 10.1128/jvi.42.3.969-977.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Mak I., Mak S. Transformation of rat cells by cyt mutants of adenovirus type 12 and mutants of adenovirus type 5. J Virol. 1983 Mar;45(3):1107–1117. doi: 10.1128/jvi.45.3.1107-1117.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. McKinnon R. D., Bacchetti S., Graham F. L. Tn5 mutagenesis of the transforming genes of human adenovirus type 5. Gene. 1982 Jul-Aug;19(1):33–42. doi: 10.1016/0378-1119(82)90186-x. [DOI] [PubMed] [Google Scholar]
  36. Mougneau E., Birg F., Rassoulzadegan M., Cuzin F. Integration sites and sequence arrangement of SV40 DNA in a homogeneous series of transformed rat fibroblast lines. Cell. 1980 Dec;22(3):917–927. doi: 10.1016/0092-8674(80)90569-3. [DOI] [PubMed] [Google Scholar]
  37. Nevins J. R. Induction of the synthesis of a 70,000 dalton mammalian heat shock protein by the adenovirus E1A gene product. Cell. 1982 Jul;29(3):913–919. doi: 10.1016/0092-8674(82)90453-6. [DOI] [PubMed] [Google Scholar]
  38. Persson H., Katze M. G., Philipson L. Purification of a native membrane-associated adenovirus tumor antigen. J Virol. 1982 Jun;42(3):905–917. doi: 10.1128/jvi.42.3.905-917.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Raska K., Jr, Dougherty J., Gallimore P. H. Product of adenovirus type 2 early gene block E1 in transformed cells elicits cytolytic response in syngeneic rats. Virology. 1982 Mar;117(2):530–535. doi: 10.1016/0042-6822(82)90495-0. [DOI] [PubMed] [Google Scholar]
  40. Raska K., Jr, Morrongiello M. P., Föhring B. Adenovirus type-12 tumor antigen. III. Tumorigenicity and immune response to syngeneic rat cells transformed with virions and isolated transforming fragment of adenovirus 12 DNA. Int J Cancer. 1980 Jul 15;26(1):79–86. doi: 10.1002/ijc.2910260113. [DOI] [PubMed] [Google Scholar]
  41. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  42. Sarnow P., Ho Y. S., Williams J., Levine A. J. Adenovirus E1b-58kd tumor antigen and SV40 large tumor antigen are physically associated with the same 54 kd cellular protein in transformed cells. Cell. 1982 Feb;28(2):387–394. doi: 10.1016/0092-8674(82)90356-7. [DOI] [PubMed] [Google Scholar]
  43. Sarnow P., Sullivan C. A., Levine A. J. A monoclonal antibody detecting the adenovirus type 5-E1b-58Kd tumor antigen: characterization of the E1b-58Kd tumor antigen in adenovirus-infected and -transformed cells. Virology. 1982 Jul 30;120(2):510–517. doi: 10.1016/0042-6822(82)90054-x. [DOI] [PubMed] [Google Scholar]
  44. Shiroki K., Shimojo H., Maeta Y., Hamada C. Tumor-specific transplantation and surface antigen in cells transformed by the adenovirus 12 DNA fragments. Virology. 1979 Nov;99(1):188–191. doi: 10.1016/0042-6822(79)90053-9. [DOI] [PubMed] [Google Scholar]
  45. Stillman B. W., White E., Grodzicker T. Independent mutations in Ad2ts111 cause degradation of cellular DNA and defective viral DNA replication. J Virol. 1984 May;50(2):598–605. doi: 10.1128/jvi.50.2.598-605.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Takemori N., Riggs J. L., Aldrich C. D. Genetic studies with tumorigenic adenoviruses. II. Heterogeneity of cyt mutants of adenovirus type 12. Virology. 1969 May;38(1):8–15. doi: 10.1016/0042-6822(69)90122-6. [DOI] [PubMed] [Google Scholar]
  47. Takemori N., Riggs J. L., Aldrich C. Genetic studies with tumorigenic adenoviruses. I. Isolation of cytocidal (cyt) mutants of adenovirus type 12. Virology. 1968 Dec;36(4):575–586. doi: 10.1016/0042-6822(68)90189-x. [DOI] [PubMed] [Google Scholar]
  48. 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]

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