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. 1988 Sep;62(9):3265–3273. doi: 10.1128/jvi.62.9.3265-3273.1988

Adenovirus type 12 E1B 19-kilodalton protein is not required for oncogenic transformation in rats.

C Edbauer 1, C Lamberti 1, J Tong 1, J Williams 1
PMCID: PMC253446  PMID: 2969983

Abstract

The adenovirus type 12 mutants in700 and pm700 carry site-specific mutations within the reading frame encoding the E1B 19-kilodalton protein (19K protein) which prevent the production of the intact 19K protein. In cultures of human A549 cells, these mutants grow just as well as the wild-type virus does, but they display a large-plaque (lp), cytocidal (cyt) phenotype. DNA in these infected cells is not degraded, but at late times in human KB cells infected by the mutants, the mutants display a DNA degradation (deg) phenotype. The transformation phenotype of these mutants is also host range. Although the mutants are defective for transformation of the 3Y1 rat cell line, they transform rat and mouse primary kidney cells in vitro at wild-type efficiency and are capable of inducing tumors in rats. These results support the view that the type 12 E1B 19K protein is not obligatory for oncogenic transformation.

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Selected References

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  1. Babiss L. E., Fisher P. B., Ginsberg H. S. Effect on transformation of mutations in the early region 1b-encoded 21- and 55-kilodalton proteins of adenovirus 5. J Virol. 1984 Nov;52(2):389–395. doi: 10.1128/jvi.52.2.389-395.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barker D. D., Berk A. J. Adenovirus proteins from both E1B reading frames are required for transformation of rodent cells by viral infection and DNA transfection. Virology. 1987 Jan;156(1):107–121. doi: 10.1016/0042-6822(87)90441-7. [DOI] [PubMed] [Google Scholar]
  3. Bernards R., de Leeuw M. G., Houweling A., van der Eb A. J. Role of the adenovirus early region 1B tumor antigens in transformation and lytic infection. Virology. 1986 Apr 15;150(1):126–139. doi: 10.1016/0042-6822(86)90272-2. [DOI] [PubMed] [Google Scholar]
  4. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Breiding D. E., Edbauer C. A., Tong J. Y., Byrd P., Grand R. J., Gallimore P. H., Williams J. Isolation and characterization of adenovirus type 12 E1 host-range mutants defective for growth in nontransformed human cells. Virology. 1988 Jun;164(2):390–402. doi: 10.1016/0042-6822(88)90552-1. [DOI] [PubMed] [Google Scholar]
  7. Byrd P. J., Chia W., Rigby P. W., Gallimore P. H. Cloning of DNA fragments from the left end of the adenovirus type 12 genome: transformation by cloned early region 1. J Gen Virol. 1982 Jun;60(Pt 2):279–293. doi: 10.1099/0022-1317-60-2-279. [DOI] [PubMed] [Google Scholar]
  8. Byrd P. J., Grand R. J., Breiding D., Williams J. F., Gallimore P. H. Host range mutants of adenovirus type 12 E1 defective for lytic infection, transformation, and oncogenicity. Virology. 1988 Mar;163(1):155–165. doi: 10.1016/0042-6822(88)90242-5. [DOI] [PubMed] [Google Scholar]
  9. Byrd P., Brown K. W., Gallimore P. H. Malignant transformation of human embryo retinoblasts by cloned adenovirus 12 DNA. Nature. 1982 Jul 1;298(5869):69–71. doi: 10.1038/298069a0. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. 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]
  12. Frost E., Williams J. Mapping temperature-sensitive and host-range mutations of adenovirus type 5 by marker rescue. Virology. 1978 Nov;91(1):39–50. doi: 10.1016/0042-6822(78)90353-7. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Fukui Y., Shiroki K., Saito I., Shimojo H. Characterization of a host range mutant of human adenovirus 12 defective in early region 1B. J Virol. 1984 Apr;50(1):132–136. doi: 10.1128/jvi.50.1.132-136.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Föhring B., Gallimore P. H., Mellow G. H., Raska K., Jr Adenovirus type 12 specific cell surface antigen in transformed cells is a product of the E1b early region. Virology. 1983 Dec;131(2):463–472. doi: 10.1016/0042-6822(83)90512-3. [DOI] [PubMed] [Google Scholar]
  16. Giard D. J., Aaronson S. A., Todaro G. J., Arnstein P., Kersey J. H., Dosik H., Parks W. P. In vitro cultivation of human tumors: establishment of cell lines derived from a series of solid tumors. J Natl Cancer Inst. 1973 Nov;51(5):1417–1423. doi: 10.1093/jnci/51.5.1417. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  19. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  20. Grand R. J., Gallimore P. H. Adenovirus type 12 early region 1 proteins: a study of their subcellular localization and protein-protein interactions. J Gen Virol. 1984 Dec;65(Pt 12):2149–2166. doi: 10.1099/0022-1317-65-12-2149. [DOI] [PubMed] [Google Scholar]
  21. Grand R. J., Roberts C., Gallimore P. H. Acylation of adenovirus type 12 early region 1b 18-kDa protein. Further evidence for its localisation in the cell membrane. FEBS Lett. 1985 Feb 25;181(2):229–235. doi: 10.1016/0014-5793(85)80265-9. [DOI] [PubMed] [Google Scholar]
  22. HUEBNER R. J., ROWE W. P., LANE W. T. Oncogenic effects in hamsters of human adenovirus types 12 and 18. Proc Natl Acad Sci U S A. 1962 Dec 15;48:2051–2058. doi: 10.1073/pnas.48.12.2051. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. 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]
  26. 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]
  27. Kimura G., Itagaki A., Summers J. Rat cell line 3y1 and its virogenic polyoma- and sv40- transformed derivatives. Int J Cancer. 1975 Apr 15;15(4):694–706. doi: 10.1002/ijc.2910150419. [DOI] [PubMed] [Google Scholar]
  28. Kozak M. Regulation of protein synthesis in virus-infected animal cells. Adv Virus Res. 1986;31:229–292. doi: 10.1016/S0065-3527(08)60265-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. 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]
  31. 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]
  32. Montell C., Fisher E. F., Caruthers M. H., Berk A. J. Resolving the functions of overlapping viral genes by site-specific mutagenesis at a mRNA splice site. Nature. 1982 Feb 4;295(5848):380–384. doi: 10.1038/295380a0. [DOI] [PubMed] [Google Scholar]
  33. Natarajan V. Adenovirus-2 E1a and E1b gene products regulate enhancer mediated transcription. Nucleic Acids Res. 1986 Dec 9;14(23):9445–9456. doi: 10.1093/nar/14.23.9445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
  35. 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]
  36. Pettersson U., Mathews M. B. The gene and messenger RNA for adenovirus polypeptide IX. Cell. 1977 Nov;12(3):741–750. doi: 10.1016/0092-8674(77)90274-4. [DOI] [PubMed] [Google Scholar]
  37. Pilder S., Logan J., Shenk T. 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. J Virol. 1984 Nov;52(2):664–671. doi: 10.1128/jvi.52.2.664-671.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. RABSON A. S., KIRSCHSTEIN R. L., PAUL F. J. TUMORS PRODUCED BY ADENOVIRUS 12 IN MASTOMYS AND MICE. J Natl Cancer Inst. 1964 Jan;32:77–87. [PubMed] [Google Scholar]
  39. Ross S. R., Levine A. J., Galos R. S., Williams J., Shenk T. Early viral proteins in HeLa cells infected with adenovirus type 5 host range mutants. Virology. 1980 Jun;103(2):475–492. doi: 10.1016/0042-6822(80)90205-6. [DOI] [PubMed] [Google Scholar]
  40. Saito I., Shiroki K., Shimojo H. mRNA species and proteins of adenovirus type 12 transforming regions: identification of proteins translated from multiple coding stretches in 2.2 kb region 1B mRNA in vitro and in vivo. Virology. 1983 Jun;127(2):272–289. doi: 10.1016/0042-6822(83)90143-5. [DOI] [PubMed] [Google Scholar]
  41. 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]
  42. 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]
  43. TRENTIN J. J., YABE Y., TAYLOR G. The quest for human cancer viruses. Science. 1962 Sep 14;137(3533):835–841. doi: 10.1126/science.137.3533.835. [DOI] [PubMed] [Google Scholar]
  44. Takemori N., Cladaras C., Bhat B., Conley A. J., Wold W. S. cyt gene of adenoviruses 2 and 5 is an oncogene for transforming function in early region E1B and encodes the E1B 19,000-molecular-weight polypeptide. J Virol. 1984 Dec;52(3):793–805. doi: 10.1128/jvi.52.3.793-805.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. 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]
  46. White E., Blose S. H., Stillman B. W. Nuclear envelope localization of an adenovirus tumor antigen maintains the integrity of cellular DNA. Mol Cell Biol. 1984 Dec;4(12):2865–2875. doi: 10.1128/mcb.4.12.2865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. White E., Faha B., Stillman B. Regulation of adenovirus gene expression in human WI38 cells by an E1B-encoded tumor antigen. Mol Cell Biol. 1986 Nov;6(11):3763–3773. doi: 10.1128/mcb.6.11.3763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. White E., Stillman B. Expression of adenovirus E1B mutant phenotypes is dependent on the host cell and on synthesis of E1A proteins. J Virol. 1987 Feb;61(2):426–435. doi: 10.1128/jvi.61.2.426-435.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Wilkie N. M., Ustacelebi S., Williams J. F. Characterization of temperature-sensitive mutants of adenovirus type 5: nucleic acid synthesis. Virology. 1973 Feb;51(2):499–503. doi: 10.1016/0042-6822(73)90450-9. [DOI] [PubMed] [Google Scholar]
  50. Williams J. F. Enhancement of adenovirus plaque formation on HeLa cells by magnesium chloride. J Gen Virol. 1970 Dec;9(3):251–255. doi: 10.1099/0022-1317-9-3-251. [DOI] [PubMed] [Google Scholar]
  51. Williams J. F., Gharpure M., Ustacelebi S., McDonald S. Isolation of temperature-sensitive mutants of adenovirus type 5. J Gen Virol. 1971 May;11(2):95–101. doi: 10.1099/0022-1317-11-2-95. [DOI] [PubMed] [Google Scholar]
  52. Williams J. F., Ustacelebi S. Temperature-restricted mutants of human adenovirus type 5. In: strategy of the viral genome. Ciba Found Symp. 1971:275–290. doi: 10.1002/9780470719824.ch16. [DOI] [PubMed] [Google Scholar]
  53. Willians J. F., Young C. S., Austin P. E. Genetic analysis of human adenovirus type 5 in permissive and nonpermissive cells. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):427–437. doi: 10.1101/sqb.1974.039.01.055. [DOI] [PubMed] [Google Scholar]
  54. YABE Y., SAMPER L., BRYAN E., TAYLOR G., TRENTIN J. J. ONCOGENIC EFFECT OF HUMAN ADENOVIRUS TYPE 12, IN MICE. Science. 1964 Jan 3;143(3601):46–47. doi: 10.1126/science.143.3601.46. [DOI] [PubMed] [Google Scholar]
  55. Yoshida K., Venkatesh L., Kuppuswamy M., Chinnadurai G. Adenovirus transforming 19-kD T antigen has an enhancer-dependent trans-activation function and relieves enhancer repression mediated by viral and cellular genes. Genes Dev. 1987 Sep;1(7):645–658. doi: 10.1101/gad.1.7.645. [DOI] [PubMed] [Google Scholar]
  56. Zoller M. J., Smith M. Oligonucleotide-directed mutagenesis using M13-derived vectors: an efficient and general procedure for the production of point mutations in any fragment of DNA. Nucleic Acids Res. 1982 Oct 25;10(20):6487–6500. doi: 10.1093/nar/10.20.6487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. van Ormondt H., Galibert F. Nucleotide sequences of adenovirus DNAs. Curr Top Microbiol Immunol. 1984;110:73–142. doi: 10.1007/978-3-642-46494-2_4. [DOI] [PubMed] [Google Scholar]

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