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EMBO J. Feb 1993; 12(2): 469–478.
PMCID: PMC413230

A region in the C-terminus of adenovirus 2/5 E1a protein is required for association with a cellular phosphoprotein and important for the negative modulation of T24-ras mediated transformation, tumorigenesis and metastasis.


We have examined a series of small deletion mutants within exon 2 of the adenovirus 2/5 E1A oncogene product, the 243R protein, for immortalization, ras cooperative transformation, tumorigenesis and metastasis. Compared with wild-type 243R, various deletion mutants located between residues 193 and 243 cooperated more efficiently with ras to induce large transformed foci of less adherent cells that were tumorigenic and metastatic. However, the greatest enhancement of transformation (comparable to that obtained with a deletion of the C-terminal 67 amino acids) was observed with a mutant carrying a deletion of residues 225-238. This mutant was also more defective in immortalization. These results suggest that this 14 amino acid region may contain a function that is important for immortalization and negative modulation of tumorigenesis and metastasis. To identify cellular proteins that may associate with the exon 2-coded region of E1A (C-terminal half) and modulate its transformation potential, we constructed a chimeric gene coding for the C-terminal 68 amino acids of E1a fused to bacterial glutathione-S-transferase (GST). This fusion protein was used to purify cellular proteins that bind to the C-terminal region of E1a. A 48 kDa cellular protein doublet (designated CtBP) was found to bind specifically to the GST-E1a C-terminal fusion protein as well as to bacterially expressed full-length E1a (243R) protein. It also co-immunoprecipitated specifically with E1a. Analysis of a panel of GST-E1a C-terminal mutant proteins indicates that residues 225-238 are required for the association of E1a and CtBP, suggesting a correlation between the association of CtBP and the immortalization and transformation modulating activities of exon 2. CtBP is a phosphoprotein and the level of phosphorylation of CtBP appears to be regulated during the cell cycle, suggesting that it may play an important role during cellular proliferation.

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  • Bagchi S, Raychaudhuri P, Nevins JR. Adenovirus E1A proteins can dissociate heteromeric complexes involving the E2F transcription factor: a novel mechanism for E1A trans-activation. Cell. 1990 Aug 24;62(4):659–669. [PubMed]
  • Bagchi S, Weinmann R, Raychaudhuri P. The retinoblastoma protein copurifies with E2F-I, an E1A-regulated inhibitor of the transcription factor E2F. Cell. 1991 Jun 14;65(6):1063–1072. [PubMed]
  • Bandara LR, La Thangue NB. Adenovirus E1a prevents the retinoblastoma gene product from complexing with a cellular transcription factor. Nature. 1991 Jun 6;351(6326):494–497. [PubMed]
  • Berk AJ. Adenovirus promoters and E1A transactivation. Annu Rev Genet. 1986;20:45–79. [PubMed]
  • Chellappan SP, Hiebert S, Mudryj M, Horowitz JM, Nevins JR. The E2F transcription factor is a cellular target for the RB protein. Cell. 1991 Jun 14;65(6):1053–1061. [PubMed]
  • Cone RD, Grodzicker T, Jaramillo M. A retrovirus expressing the 12S adenoviral E1A gene product can immortalize epithelial cells from a broad range of rat tissues. Mol Cell Biol. 1988 Mar;8(3):1036–1044. [PMC free article] [PubMed]
  • Douglas JL, Gopalakrishnan S, Quinlan MP. Modulation of transformation of primary epithelial cells by the second exon of the Ad5 E1A12S gene. Oncogene. 1991 Nov;6(11):2093–2103. [PubMed]
  • Egan C, Jelsma TN, Howe JA, Bayley ST, Ferguson B, Branton PE. Mapping of cellular protein-binding sites on the products of early-region 1A of human adenovirus type 5. Mol Cell Biol. 1988 Sep;8(9):3955–3959. [PMC free article] [PubMed]
  • Frisch SM, Reich R, Collier IE, Genrich LT, Martin G, Goldberg GI. Adenovirus E1A represses protease gene expression and inhibits metastasis of human tumor cells. Oncogene. 1990 Jan;5(1):75–83. [PubMed]
  • Giordano A, McCall C, Whyte P, Franza BR., Jr Human cyclin A and the retinoblastoma protein interact with similar but distinguishable sequences in the adenovirus E1A gene product. Oncogene. 1991 Mar;6(3):481–485. [PubMed]
  • Giordano A, Lee JH, Scheppler JA, Herrmann C, Harlow E, Deuschle U, Beach D, Franza BR., Jr Cell cycle regulation of histone H1 kinase activity associated with the adenoviral protein E1A. Science. 1991 Sep 13;253(5025):1271–1275. [PubMed]
  • Harlow E, Franza BR, 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. [PMC free article] [PubMed]
  • Harlow E, Whyte P, Franza BR, Jr, Schley C. Association of adenovirus early-region 1A proteins with cellular polypeptides. Mol Cell Biol. 1986 May;6(5):1579–1589. [PMC free article] [PubMed]
  • Heintz N, Sive HL, Roeder RG. Regulation of human histone gene expression: kinetics of accumulation and changes in the rate of synthesis and in the half-lives of individual histone mRNAs during the HeLa cell cycle. Mol Cell Biol. 1983 Apr;3(4):539–550. [PMC free article] [PubMed]
  • Jelsma TN, Howe JA, Evelegh CM, Cunniff NF, Skiadopoulos MH, Floroff MR, Denman JE, Bayley ST. Use of deletion and point mutants spanning the coding region of the adenovirus 5 E1A gene to define a domain that is essential for transcriptional activation. Virology. 1988 Apr;163(2):494–502. [PubMed]
  • Jelsma TN, Howe JA, Mymryk JS, Evelegh CM, Cunniff NF, Bayley ST. Sequences in E1A proteins of human adenovirus 5 required for cell transformation, repression of a transcriptional enhancer, and induction of proliferating cell nuclear antigen. Virology. 1989 Jul;171(1):120–130. [PubMed]
  • Kaelin WG, Jr, Pallas DC, DeCaprio JA, Kaye FJ, Livingston DM. Identification of cellular proteins that can interact specifically with the T/E1A-binding region of the retinoblastoma gene product. Cell. 1991 Feb 8;64(3):521–532. [PubMed]
  • Kast WM, Offringa R, Peters PJ, Voordouw AC, Meloen RH, van der Eb AJ, Melief CJ. Eradication of adenovirus E1-induced tumors by E1A-specific cytotoxic T lymphocytes. Cell. 1989 Nov 17;59(4):603–614. [PubMed]
  • Kleinberger T, Shenk T. A protein kinase is present in a complex with adenovirus E1A proteins. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11143–11147. [PMC free article] [PubMed]
  • Kuppuswamy MN, Chinnadurai G. Relationship between the transforming and transcriptional regulatory functions of adenovirus 2 E1a oncogene. Virology. 1987 Jul;159(1):31–38. [PubMed]
  • Kuppuswamy M, Chinnadurai G. Cell type dependent transformation by adenovirus 5 E1a proteins. Oncogene. 1988 Jun;2(6):567–572. [PubMed]
  • Lillie JW, Green M, Green MR. An adenovirus E1a protein region required for transformation and transcriptional repression. Cell. 1986 Sep 26;46(7):1043–1051. [PubMed]
  • Liotta LA, Steeg PS, Stetler-Stevenson WG. Cancer metastasis and angiogenesis: an imbalance of positive and negative regulation. Cell. 1991 Jan 25;64(2):327–336. [PubMed]
  • Moran E, Zerler B, Harrison TM, Mathews MB. Identification of separate domains in the adenovirus E1A gene for immortalization activity and the activation of virus early genes. Mol Cell Biol. 1986 Oct;6(10):3470–3480. [PMC free article] [PubMed]
  • Mudryj M, Hiebert SW, Nevins JR. A role for the adenovirus inducible E2F transcription factor in a proliferation dependent signal transduction pathway. EMBO J. 1990 Jul;9(7):2179–2184. [PMC free article] [PubMed]
  • Offringa R, Gebel S, van Dam H, Timmers M, Smits A, Zwart R, Stein B, Bos JL, van der Eb A, Herrlich P. A novel function of the transforming domain of E1a: repression of AP-1 activity. Cell. 1990 Aug 10;62(3):527–538. [PubMed]
  • Pines J, Hunter T. Human cyclin A is adenovirus E1A-associated protein p60 and behaves differently from cyclin B. Nature. 1990 Aug 23;346(6286):760–763. [PubMed]
  • Quinlan MP, Douglas JL. Immortalization of primary epithelial cells requires first- and second-exon functions of adenovirus type 5 12S. J Virol. 1992 Apr;66(4):2020–2030. [PMC free article] [PubMed]
  • Quinlan MP, Whyte P, Grodzicker T. Growth factor induction by the adenovirus type 5 E1A 12S protein is required for immortalization of primary epithelial cells. Mol Cell Biol. 1988 Aug;8(8):3191–3203. [PMC free article] [PubMed]
  • Raychaudhuri P, Bagchi S, Devoto SH, Kraus VB, Moran E, Nevins JR. Domains of the adenovirus E1A protein required for oncogenic activity are also required for dissociation of E2F transcription factor complexes. Genes Dev. 1991 Jul;5(7):1200–1211. [PubMed]
  • Ruley HE. Transforming collaborations between ras and nuclear oncogenes. Cancer Cells. 1990 Aug-Sep;2(8-9):258–268. [PubMed]
  • Schneider JF, Fisher F, Goding CR, Jones NC. Mutational analysis of the adenovirus E1a gene: the role of transcriptional regulation in transformation. EMBO J. 1987 Jul;6(7):2053–2060. [PMC free article] [PubMed]
  • Smith DB, Johnson KS. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. [PubMed]
  • Smith DH, Ziff EB. The amino-terminal region of the adenovirus serotype 5 E1a protein performs two separate functions when expressed in primary baby rat kidney cells. Mol Cell Biol. 1988 Sep;8(9):3882–3890. [PMC free article] [PubMed]
  • Steeg PS, Bevilacqua G, Pozzatti R, Liotta LA, Sobel ME. Altered expression of NM23, a gene associated with low tumor metastatic potential, during adenovirus 2 Ela inhibition of experimental metastasis. Cancer Res. 1988 Nov 15;48(22):6550–6554. [PubMed]
  • Stein RW, Corrigan M, Yaciuk P, Whelan J, Moran E. Analysis of E1A-mediated growth regulation functions: binding of the 300-kilodalton cellular product correlates with E1A enhancer repression function and DNA synthesis-inducing activity. J Virol. 1990 Sep;64(9):4421–4427. [PMC free article] [PubMed]
  • Subramanian T, Kuppuswamy M, Nasr RJ, Chinnadurai G. An N-terminal region of adenovirus E1a essential for cell transformation and induction of an epithelial cell growth factor. Oncogene. 1988 Feb;2(2):105–112. [PubMed]
  • Subramanian T, La Regina M, Chinnadurai G. Enhanced ras oncogene mediated cell transformation and tumorigenesis by adenovirus 2 mutants lacking the C-terminal region of E1a protein. Oncogene. 1989 Apr;4(4):415–420. [PubMed]
  • Tsai LH, Harlow E, Meyerson M. Isolation of the human cdk2 gene that encodes the cyclin A- and adenovirus E1A-associated p33 kinase. Nature. 1991 Sep 12;353(6340):174–177. [PubMed]
  • Urbanelli D, Sawada Y, Raskova J, Jones NC, Shenk T, Raska K., Jr C-terminal domain of the adenovirus E1A oncogene product is required for induction of cytotoxic T lymphocytes and tumor-specific transplantation immunity. Virology. 1989 Dec;173(2):607–614. [PubMed]
  • Whyte P, Buchkovich KJ, Horowitz JM, Friend SH, Raybuck M, Weinberg RA, Harlow E. Association between an oncogene and an anti-oncogene: the adenovirus E1A proteins bind to the retinoblastoma gene product. Nature. 1988 Jul 14;334(6178):124–129. [PubMed]
  • Whyte P, Williamson NM, Harlow E. Cellular targets for transformation by the adenovirus E1A proteins. Cell. 1989 Jan 13;56(1):67–75. [PubMed]
  • Yee SP, Branton PE. Detection of cellular proteins associated with human adenovirus type 5 early region 1A polypeptides. Virology. 1985 Nov;147(1):142–153. [PubMed]
  • Yu DH, Scorsone K, Hung MC. Adenovirus type 5 E1A gene products act as transformation suppressors of the neu oncogene. Mol Cell Biol. 1991 Mar;11(3):1745–1750. [PMC free article] [PubMed]
  • Zerler B, Moran B, Maruyama K, Moomaw J, Grodzicker T, Ruley HE. Adenovirus E1A coding sequences that enable ras and pmt oncogenes to transform cultured primary cells. Mol Cell Biol. 1986 Mar;6(3):887–899. [PMC free article] [PubMed]
  • Zerler B, Roberts RJ, Mathews MB, Moran E. Different functional domains of the adenovirus E1A gene are involved in regulation of host cell cycle products. Mol Cell Biol. 1987 Feb;7(2):821–829. [PMC free article] [PubMed]

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