Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1997 May;71(5):3788–3798. doi: 10.1128/jvi.71.5.3788-3798.1997

Regulation of p53 levels by the E1B 55-kilodalton protein and E4orf6 in adenovirus-infected cells.

E Querido 1, R C Marcellus 1, A Lai 1, R Charbonneau 1, J G Teodoro 1, G Ketner 1, P E Branton 1
PMCID: PMC191529  PMID: 9094654

Abstract

The adenovirus type 5 243R E1A protein induces p53-dependent apoptosis in the absence of the 19- and 55-kDa E1B polypeptides. This effect appears to result from an accumulation of p53 protein and is unrelated to expression of E1B products. We now report that in the presence of the E1B 55-kDa polypeptide, the 289R E1A protein does not induce such p53 accumulation and, in fact, is able to block that induced by E1A 243R. This inhibition also requires the 289R-dependent transactivation of E4orf6 expression. E4orf6 is known to form complexes with the E1B 55-kDa protein and to function both in the transport and stabilization of viral mRNA and in shutoff of host cell protein synthesis. We demonstrated that the block in p53 accumulation is not due to the generalized shutoff of host cell metabolism. Rather, it appears to result from a mechanism targeted specifically to p53, most likely involving a decrease in the stability of p53 protein. The E1B 55-kDa protein is known to interact with both E4orf6 and p53, and as demonstrated recently by others, we showed that E4orf6 also binds directly to p53. Thus, multiple interactions between all three proteins may regulate p53 stability, resulting in the maintenance of low levels of p53 following virus infection.

Full Text

The Full Text of this article is available as a PDF (2.8 MB).

Selected References

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

  1. 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]
  2. Babiss L. E., Ginsberg H. S., Darnell J. E., Jr Adenovirus E1B proteins are required for accumulation of late viral mRNA and for effects on cellular mRNA translation and transport. Mol Cell Biol. 1985 Oct;5(10):2552–2558. doi: 10.1128/mcb.5.10.2552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boyd J. M., Malstrom S., Subramanian T., Venkatesh L. K., Schaeper U., Elangovan B., D'Sa-Eipper C., Chinnadurai G. Adenovirus E1B 19 kDa and Bcl-2 proteins interact with a common set of cellular proteins. Cell. 1994 Oct 21;79(2):341–351. doi: 10.1016/0092-8674(94)90202-x. [DOI] [PubMed] [Google Scholar]
  4. Braithwaite A., Nelson C., Skulimowski A., McGovern J., Pigott D., Jenkins J. Transactivation of the p53 oncogene by E1a gene products. Virology. 1990 Aug;177(2):595–605. doi: 10.1016/0042-6822(90)90525-v. [DOI] [PubMed] [Google Scholar]
  5. Bridge E., Ketner G. Interaction of adenoviral E4 and E1b products in late gene expression. Virology. 1990 Feb;174(2):345–353. doi: 10.1016/0042-6822(90)90088-9. [DOI] [PubMed] [Google Scholar]
  6. Bridge E., Ketner G. Redundant control of adenovirus late gene expression by early region 4. J Virol. 1989 Feb;63(2):631–638. doi: 10.1128/jvi.63.2.631-638.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bridge E., Medghalchi S., Ubol S., Leesong M., Ketner G. Adenovirus early region 4 and viral DNA synthesis. Virology. 1993 Apr;193(2):794–801. doi: 10.1006/viro.1993.1188. [DOI] [PubMed] [Google Scholar]
  8. Chiou S. K., Tseng C. C., Rao L., White E. Functional complementation of the adenovirus E1B 19-kilodalton protein with Bcl-2 in the inhibition of apoptosis in infected cells. J Virol. 1994 Oct;68(10):6553–6566. doi: 10.1128/jvi.68.10.6553-6566.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Crook T., Wrede D., Vousden K. H. p53 point mutation in HPV negative human cervical carcinoma cell lines. Oncogene. 1991 May;6(5):873–875. [PubMed] [Google Scholar]
  10. Cutt J. R., Shenk T., Hearing P. Analysis of adenovirus early region 4-encoded polypeptides synthesized in productively infected cells. J Virol. 1987 Feb;61(2):543–552. doi: 10.1128/jvi.61.2.543-552.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Debbas M., White E. Wild-type p53 mediates apoptosis by E1A, which is inhibited by E1B. Genes Dev. 1993 Apr;7(4):546–554. doi: 10.1101/gad.7.4.546. [DOI] [PubMed] [Google Scholar]
  12. Dobner T., Horikoshi N., Rubenwolf S., Shenk T. Blockage by adenovirus E4orf6 of transcriptional activation by the p53 tumor suppressor. Science. 1996 Jun 7;272(5267):1470–1473. doi: 10.1126/science.272.5267.1470. [DOI] [PubMed] [Google Scholar]
  13. Fu L., Minden M. D., Benchimol S. Translational regulation of human p53 gene expression. EMBO J. 1996 Aug 15;15(16):4392–4401. [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Grand R. J., Grant M. L., Gallimore P. H. Enhanced expression of p53 in human cells infected with mutant adenoviruses. Virology. 1994 Sep;203(2):229–240. doi: 10.1006/viro.1994.1480. [DOI] [PubMed] [Google Scholar]
  16. Grand R. J., Lecane P. S., Owen D., Grant M. L., Roberts S., Levine A. J., Gallimore P. H. The high levels of p53 present in adenovirus early region 1-transformed human cells do not cause up-regulation of MDM2 expression. Virology. 1995 Jul 10;210(2):323–334. doi: 10.1006/viro.1995.1349. [DOI] [PubMed] [Google Scholar]
  17. Grand R. J., Lecane P. S., Roberts S., Grant M. L., Lane D. P., Young L. S., Dawson C. W., Gallimore P. H. Overexpression of wild-type p53 and c-Myc in human fetal cells transformed with adenovirus early region 1. Virology. 1993 Apr;193(2):579–591. doi: 10.1006/viro.1993.1166. [DOI] [PubMed] [Google Scholar]
  18. Grand R. J., Owen D., Rookes S. M., Gallimore P. H. Control of p53 expression by adenovirus 12 early region 1A and early region 1B 54K proteins. Virology. 1996 Apr 1;218(1):23–34. doi: 10.1006/viro.1996.0162. [DOI] [PubMed] [Google Scholar]
  19. Halbert D. N., Cutt J. R., Shenk T. Adenovirus early region 4 encodes functions required for efficient DNA replication, late gene expression, and host cell shutoff. J Virol. 1985 Oct;56(1):250–257. doi: 10.1128/jvi.56.1.250-257.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Haley K. P., Overhauser J., Babiss L. E., Ginsberg H. S., Jones N. C. Transformation properties of type 5 adenovirus mutants that differentially express the E1A gene products. Proc Natl Acad Sci U S A. 1984 Sep;81(18):5734–5738. doi: 10.1073/pnas.81.18.5734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Harlow E., Franza B. R., Jr, Schley C. Monoclonal antibodies specific for adenovirus early region 1A proteins: extensive heterogeneity in early region 1A products. J Virol. 1985 Sep;55(3):533–546. doi: 10.1128/jvi.55.3.533-546.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. Howe J. A., Bayley S. T. Effects of Ad5 E1A mutant viruses on the cell cycle in relation to the binding of cellular proteins including the retinoblastoma protein and cyclin A. Virology. 1992 Jan;186(1):15–24. doi: 10.1016/0042-6822(92)90057-v. [DOI] [PubMed] [Google Scholar]
  24. Howe J. A., Mymryk J. S., Egan C., Branton P. E., Bayley S. T. Retinoblastoma growth suppressor and a 300-kDa protein appear to regulate cellular DNA synthesis. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5883–5887. doi: 10.1073/pnas.87.15.5883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Kao C. C., Yew P. R., Berk A. J. Domains required for in vitro association between the cellular p53 and the adenovirus 2 E1B 55K proteins. Virology. 1990 Dec;179(2):806–814. doi: 10.1016/0042-6822(90)90148-k. [DOI] [PubMed] [Google Scholar]
  27. Ko L. J., Prives C. p53: puzzle and paradigm. Genes Dev. 1996 May 1;10(9):1054–1072. doi: 10.1101/gad.10.9.1054. [DOI] [PubMed] [Google Scholar]
  28. Leppard K. N., Shenk T. The adenovirus E1B 55 kd protein influences mRNA transport via an intranuclear effect on RNA metabolism. EMBO J. 1989 Aug;8(8):2329–2336. doi: 10.1002/j.1460-2075.1989.tb08360.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Linzer D. I., Levine A. J. Characterization of a 54K dalton cellular SV40 tumor antigen present in SV40-transformed cells and uninfected embryonal carcinoma cells. Cell. 1979 May;17(1):43–52. doi: 10.1016/0092-8674(79)90293-9. [DOI] [PubMed] [Google Scholar]
  30. Lowe S. W., Ruley H. E., Jacks T., Housman D. E. p53-dependent apoptosis modulates the cytotoxicity of anticancer agents. Cell. 1993 Sep 24;74(6):957–967. doi: 10.1016/0092-8674(93)90719-7. [DOI] [PubMed] [Google Scholar]
  31. Lowe S. W., Ruley H. E. Stabilization of the p53 tumor suppressor is induced by adenovirus 5 E1A and accompanies apoptosis. Genes Dev. 1993 Apr;7(4):535–545. doi: 10.1101/gad.7.4.535. [DOI] [PubMed] [Google Scholar]
  32. Marcellus R. C., Teodoro J. G., Charbonneau R., Shore G. C., Branton P. E. Expression of p53 in Saos-2 osteosarcoma cells induces apoptosis which can be inhibited by Bcl-2 or the adenovirus E1B-55 kDa protein. Cell Growth Differ. 1996 Dec;7(12):1643–1650. [PubMed] [Google Scholar]
  33. Marcellus R. C., Teodoro J. G., Wu T., Brough D. E., Ketner G., Shore G. C., Branton P. E. Adenovirus type 5 early region 4 is responsible for E1A-induced p53-independent apoptosis. J Virol. 1996 Sep;70(9):6207–6215. doi: 10.1128/jvi.70.9.6207-6215.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. McLorie W., McGlade C. J., Takayesu D., Branton P. E. Individual adenovirus E1B proteins induce transformation independently but by additive pathways. J Gen Virol. 1991 Jun;72(Pt 6):1467–1471. doi: 10.1099/0022-1317-72-6-1467. [DOI] [PubMed] [Google Scholar]
  35. 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]
  36. Moore M., Horikoshi N., Shenk T. Oncogenic potential of the adenovirus E4orf6 protein. Proc Natl Acad Sci U S A. 1996 Oct 15;93(21):11295–11301. doi: 10.1073/pnas.93.21.11295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Mosner J., Mummenbrauer T., Bauer C., Sczakiel G., Grosse F., Deppert W. Negative feedback regulation of wild-type p53 biosynthesis. EMBO J. 1995 Sep 15;14(18):4442–4449. doi: 10.1002/j.1460-2075.1995.tb00123.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Nguyen M., Branton P. E., Walton P. A., Oltvai Z. N., Korsmeyer S. J., Shore G. C. Role of membrane anchor domain of Bcl-2 in suppression of apoptosis caused by E1B-defective adenovirus. J Biol Chem. 1994 Jun 17;269(24):16521–16524. [PubMed] [Google Scholar]
  39. Obert S., O'Connor R. J., Schmid S., Hearing P. The adenovirus E4-6/7 protein transactivates the E2 promoter by inducing dimerization of a heteromeric E2F complex. Mol Cell Biol. 1994 Feb;14(2):1333–1346. doi: 10.1128/mcb.14.2.1333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Ohman K., Nordqvist K., Akusjärvi G. Two adenovirus proteins with redundant activities in virus growth facilitates tripartite leader mRNA accumulation. Virology. 1993 May;194(1):50–58. doi: 10.1006/viro.1993.1234. [DOI] [PubMed] [Google Scholar]
  41. Ornelles D. A., Shenk T. Localization of the adenovirus early region 1B 55-kilodalton protein during lytic infection: association with nuclear viral inclusions requires the early region 4 34-kilodalton protein. J Virol. 1991 Jan;65(1):424–429. doi: 10.1128/jvi.65.1.424-429.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Pilder S., Moore M., Logan J., Shenk T. The adenovirus E1B-55K transforming polypeptide modulates transport or cytoplasmic stabilization of viral and host cell mRNAs. Mol Cell Biol. 1986 Feb;6(2):470–476. doi: 10.1128/mcb.6.2.470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Querido E., Teodoro J. G., Branton P. E. Accumulation of p53 induced by the adenovirus E1A protein requires regions involved in the stimulation of DNA synthesis. J Virol. 1997 May;71(5):3526–3533. doi: 10.1128/jvi.71.5.3526-3533.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Rao L., Debbas M., Sabbatini P., Hockenbery D., Korsmeyer S., White E. The adenovirus E1A proteins induce apoptosis, which is inhibited by the E1B 19-kDa and Bcl-2 proteins. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7742–7746. doi: 10.1073/pnas.89.16.7742. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Reznikoff C. A., Brankow D. W., Heidelberger C. Establishment and characterization of a cloned line of C3H mouse embryo cells sensitive to postconfluence inhibition of division. Cancer Res. 1973 Dec;33(12):3231–3238. [PubMed] [Google Scholar]
  46. Rowe D. T., Branton P. E., Graham F. L. The kinetics of synthesis of early viral proteins in KB cells infected with wild-type and transformation-defective host-range mutants of human adenovirus type 5. J Gen Virol. 1984 Mar;65(Pt 3):585–597. doi: 10.1099/0022-1317-65-3-585. [DOI] [PubMed] [Google Scholar]
  47. Sanchez-Prieto R., Lleonart M., Ramón y Cajal S. Lack of correlation between p53 protein level and sensitivity of DNA-damaging agents in keratinocytes carrying adenovirus E1a mutants. Oncogene. 1995 Aug 17;11(4):675–682. [PubMed] [Google Scholar]
  48. Sandler A. B., Ketner G. Adenovirus early region 4 is essential for normal stability of late nuclear RNAs. J Virol. 1989 Feb;63(2):624–630. doi: 10.1128/jvi.63.2.624-630.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Sandler A. B., Ketner G. The metabolism of host RNAs in cells infected by an adenovirus E4 mutant. Virology. 1991 Mar;181(1):319–326. doi: 10.1016/0042-6822(91)90498-z. [DOI] [PubMed] [Google Scholar]
  50. Sarnow P., Hearing P., Anderson C. W., Halbert D. N., Shenk T., Levine A. J. Adenovirus early region 1B 58,000-dalton tumor antigen is physically associated with an early region 4 25,000-dalton protein in productively infected cells. J Virol. 1984 Mar;49(3):692–700. doi: 10.1128/jvi.49.3.692-700.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. 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]
  52. 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]
  53. Shenk T., Flint J. Transcriptional and transforming activities of the adenovirus E1A proteins. Adv Cancer Res. 1991;57:47–85. doi: 10.1016/s0065-230x(08)60995-1. [DOI] [PubMed] [Google Scholar]
  54. Shepherd S. E., Howe J. A., Mymryk J. S., Bayley S. T. Induction of the cell cycle in baby rat kidney cells by adenovirus type 5 E1A in the absence of E1B and a possible influence of p53. J Virol. 1993 May;67(5):2944–2949. doi: 10.1128/jvi.67.5.2944-2949.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Subramanian T., Tarodi B., Chinnadurai G. p53-independent apoptotic and necrotic cell deaths induced by adenovirus infection: suppression by E1B 19K and Bcl-2 proteins. Cell Growth Differ. 1995 Feb;6(2):131–137. [PubMed] [Google Scholar]
  56. Teodoro J. G., Branton P. E. Regulation of p53-dependent apoptosis, transcriptional repression, and cell transformation by phosphorylation of the 55-kilodalton E1B protein of human adenovirus type 5. J Virol. 1997 May;71(5):3620–3627. doi: 10.1128/jvi.71.5.3620-3627.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Teodoro J. G., Halliday T., Whalen S. G., Takayesu D., Graham F. L., Branton P. E. Phosphorylation at the carboxy terminus of the 55-kilodalton adenovirus type 5 E1B protein regulates transforming activity. J Virol. 1994 Feb;68(2):776–786. doi: 10.1128/jvi.68.2.776-786.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Teodoro J. G., Shore G. C., Branton P. E. Adenovirus E1A proteins induce apoptosis by both p53-dependent and p53-independent mechanisms. Oncogene. 1995 Aug 3;11(3):467–474. [PubMed] [Google Scholar]
  59. Weinberg D. H., Ketner G. A cell line that supports the growth of a defective early region 4 deletion mutant of human adenovirus type 2. Proc Natl Acad Sci U S A. 1983 Sep;80(17):5383–5386. doi: 10.1073/pnas.80.17.5383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Weinberg D. H., Ketner G. Adenoviral early region 4 is required for efficient viral DNA replication and for late gene expression. J Virol. 1986 Mar;57(3):833–838. doi: 10.1128/jvi.57.3.833-838.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Yew P. R., Berk A. J. Inhibition of p53 transactivation required for transformation by adenovirus early 1B protein. Nature. 1992 May 7;357(6373):82–85. doi: 10.1038/357082a0. [DOI] [PubMed] [Google Scholar]
  62. Yew P. R., Kao C. C., Berk A. J. Dissection of functional domains in the adenovirus 2 early 1B 55K polypeptide by suppressor-linker insertional mutagenesis. Virology. 1990 Dec;179(2):795–805. doi: 10.1016/0042-6822(90)90147-j. [DOI] [PubMed] [Google Scholar]
  63. Yew P. R., Liu X., Berk A. J. Adenovirus E1B oncoprotein tethers a transcriptional repression domain to p53. Genes Dev. 1994 Jan;8(2):190–202. doi: 10.1101/gad.8.2.190. [DOI] [PubMed] [Google Scholar]
  64. Zantema A., Schrier P. I., Davis-Olivier A., van Laar T., Vaessen R. T., van der EB A. J. Adenovirus serotype determines association and localization of the large E1B tumor antigen with cellular tumor antigen p53 in transformed cells. Mol Cell Biol. 1985 Nov;5(11):3084–3091. doi: 10.1128/mcb.5.11.3084. [DOI] [PMC free article] [PubMed] [Google Scholar]
  65. van den Heuvel S. J., van Laar T., Kast W. M., Melief C. J., Zantema A., van der Eb A. J. Association between the cellular p53 and the adenovirus 5 E1B-55kd proteins reduces the oncogenicity of Ad-transformed cells. EMBO J. 1990 Aug;9(8):2621–2629. doi: 10.1002/j.1460-2075.1990.tb07444.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES