Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Paper
  • Published:

Exon 4-encoded acidic domain in the epithelium-restricted Ets factor, ESX, confers potent transactivating capacity and binds to TATA-binding protein (TBP)

Oncogene volume 18, pages 3682–3695 (1999)Cite this article

Abstract

The Ets gene family has a complex evolutionary history with many family members known to regulate genetic programs essential for differentiation and development, and some known for their involvement in human tumorigenesis. To understand the biological properties associated with a recently described epithelium-restricted Ets factor ESX, an 11 kb fragment from the 1q32.2 genomically localized human gene was cloned and analysed. Upstream of the ESX promoter region in this genomic fragment lies the terminal exon of a newly identified gene that encodes a ubiquitin-conjugating enzyme variant, UEV-1. Tissues expressing ESX produce a primary 2.2 kb transcript along with a 4.1 kb secondary transcript arising by alternate poly(A) site selection and uniquely recognized by a genomic probe from the 3′ terminal region of the 11 kb clone. Endogenous expression of ESX results in a 42 kDa nuclear protein having fivefold greater affinity for the chromatin-nuclear matrix compartment as compared to other endogenous transcription factors like AP-2 and the homologous Ets factor, ELF-1. Exon mapping of the modular structure inferred from ESX cDNA and construction of GAL4(DBD)-ESX expression constructs were used to identify a transactivating domain encoded by exon 4 having comparable potency to the acidic transactivation domain of the viral transcription factor, VP16. This exon 4-encoded 31 amino acid domain in ESX was shown by mutation and deletion analysis to possess a 13 residue acidic transactivation core which, based on modeling and circular dichroism analysis, is predicted to form an amphipathic α-helical secondary structure. Using recombinant GST-ESX (exon 4) fusion proteins in an in vitro pull-down assay, this ESX transactivation domain was shown to bind specifically to one component of the general transcription machinery, TATA-binding protein (TBP). Transient transfection experiments confirmed the ability of this TBP-binding transactivation domain in ESX to squelch heterologous promoters independent of any promoter binding as efficiently as the transactivation domain from VP16.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

Similar content being viewed by others

References

  • Andreoli JM, Jang SI, Chung E, Coticchia CM, Steinert PM and Markova NG. . 1997 Nucleic Acids Res. 25: 4287–4295.

  • Ashar HR, Fejzo MS, Tkachenko A, Zhou X, Fletcher JA, Weremowicz S, Morton CC and Chada K. . 1995 Cell 82: 57–65.

  • Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA and Struhl K. . 1989 Current Protocols in Molecular Biology John Wiley & Sons: New York.

    Google Scholar 

  • Baens M, Peeters P, Guo CY, Aerssens J and Marynen P. . 1996 Genome Res. 6: 404–413.

  • Bellacosa A, Datta K, Bear SE, Patriotis C, Lazo PA, Copeland NG, Jenkins NA and Tsichlis PN. . 1994 J. Virol. 68: 2320–2330.

  • Bochert MA, Kleinbaum LA, Sun LY and Burton FH. . 1998 Biochem. Biophys. Res. Comm. 246: 176–181.

  • Chang C and Gralla JD. . 1993 Mol. Cell. Biol. 13: 7469–7475.

  • Chang C-H, Scott GK, Kuo WL, Xiong X, Suzdaltseva Y, Park JW, Sayre P, Erny K, Collins C, Gray JW and Benz CC. . 1997 Oncogene 14: 1617–1622.

  • Chen HM, Gonzalez DA, Radomska HS, Voso MT, Sun Z, Zhang P, Zhang DE and Tenen DG. . 1998 Gene 207: 209–218.

  • Choi SG, Yi Y, Kim YS, Kato M, Chang J, Chung HW, Hahm KB, Yang HK, Rhee HH, Bang YJ and Kim SJ. . 1998 J. Biol. Chem. 273: 110–117.

  • Chou PY and Fasman GD. . 1978 Ann. Rev. Biochem. 47: 251–276.

  • Crepieux P, Coll J and Stehelin D. . 1994 Crit. Rev. Oncogenesis 5: 615–638.

  • de Castro CM, Rabe SM, Langdon SD, Fleenor DE, SlentzKesler K, Ahmed MN, Qumsiyeh MB and Kaufman RE. . 1997 Genomics 42: 227–235.

  • Defossez PA, Baert JL, Monnot M and deLaunoit Y. . 1997 Nucleic Acids Res. 25: 4455–4463.

  • Dittmer J and Nordheim A. . 1998 Biochim. Biophys. Acta 1377: F1–11.

  • Grevin D, Ung S, Denhez F, Dehem M, Quatannens B, Begue A, Stehelin D and Martin P. . 1996 Gene 174: 185–188.

  • Hromas R and Klemsz M. . 1994 Int. J. Hematol. 59: 257–265.

  • Johnson Jr WC. . 1990 Proteins 7: 205–214.

  • Jorcyk CL, Watson DK, Mavrothalassitis GJ and Papas TS. . 1991 Oncogene 6: 523–532.

  • Klambt C. . 1993 Development 117: 163–176.

  • Liu D, Pavlopoulos E, Modi W, Moschonas N and Mavrothalassitis G. . 1997 Oncogene 14: 1445–1451.

  • Monte D, Coutte L, Dewitte F, Defossez PA, Leconiat M, Stehelin D, Berger R and Delaunoit Y. . 1996 Genomics 35: 236–240.

  • Moreau-Gachelin F, Ray D, Mattei MG, Tambourin P and Tavitian A. . 1989 Oncogene 4: 1449–1456.

  • Natesan S, Rivera VM, Molinari E and Gilman M. . 1997 Nature 390: 349–350.

  • Neve R, Chang C-H, Scott GK, Wong A, Friis RR, Hynes NE and Benz CC. . 1998 FASEB J. 12: 1541–1550.

  • Oelgeschläger M, Krieg J, Lüscher-Firzlaff JM and Lüscher B. . 1995 Mol. Cell. Biol. 15: 5966–5974.

  • Oettgen P, Alani RM, Barcinski MA, Brown L, Akbarali Y, Boltax J, Kunsch C, Munger K and Libermann TA. . 1997a Mol. Cell. Biol. 17: 4419–4433.

  • Oettgen P, Carter KC, Augustus M, Barcinski M, Boltax J, Kunsch C and Libermann TA. . 1997b Genomics 45: 456–457.

  • Pongubala JM, Van Beveren C, Nagulapalli S, Klemsz MJ, McKercher SR, Maki RA and Atchison ML. . 1993 Science 259: 1622–1625.

  • Pribyl LJ, Watson DK, McWilliams MJ, Ascione R and Papas TS. . 1988 Dev. Biol. 127: 45–53.

  • Ptashne M and Gann A. . 1997 Nature 386: 569–577.

  • Pugh BF. . 1997 Transcription Factors in Eukaryotes. Papavassiliou AG (ed.).. Springer-Verlag: Heidelberg, Germany pp. 37–50.

    Google Scholar 

  • Raziuddin A, Court D, Sarkar FH, Liu YL, Kung H and Raziuddin R. . 1997 J. Biol. Chem. 272: 15715–15720.

  • Robbins J, Dilworth SM, Laskey RA and Dingwall C. . 1991 Cell 64: 615–623.

  • Sambrook J, Fritsch EF and Maniatis T. . 1989 Molecular Cloning. Cold Spring Harbor Laboratory Press: New York.

    Google Scholar 

  • Sancho E, Vila MR, Sanchez-Pulido L, Lozano JJ, Paciucci R, Nadal M, Fox M, Harvey C, Bercovich B, Loukili N, Ciechanover A, Lin SL, Sanz F, Estivill X, Valencia A and Thomson TM. . 1998 Mol. Cell. Biol. 18: 576–589.

  • Schmitz ML, dos Santos Silva MA, Altmann H, Czisch M, Holak TA and Baeuerle PA. . 1994 J. Biol. Chem. 269: 25613–25620.

  • Schneikert J, Lutz Y and Wasylyk B. . 1992 Oncogene 7: 249–256.

  • Sharrocks AD, Brown AL, Ling Y and Yates PR. . 1997 Int. J. Biochem. Cell Biol. 29: 1371–1387.

  • Tymms MJ, Ng AYN, Thomas RS, Schutte BC, Zhou JO, Eyre HJ, Sutherland GR, Seth A, Rosenberg M, Papas T, Debouck C and Kola I. . 1997 Oncogene 15: 2449–2462.

  • Uesugi M, Nyanguile O, Lu H, Levine AJ and Verdine GL. . 1997 Science 277: 1310–1313.

  • Van Hoy M, Leuther KK, Kodadek T and Johnston SA. . 1993 Cell 72: 587–594.

  • Van Wijnen AJ, Bidwell JP, Fey EG, Penman S, Lian JB, Stein JL and Stein GS. . 1993 Biochemistry 32: 8397–8402.

  • Wasylyk B and Nordheim A. . 1997 Transcription Factors in Eukaryotes. Papavassiliou AG (ed.).. Springer-Verlag: Heidelberg, Germany pp. 253–286.

    Google Scholar 

  • Watson DK, Mavrothalassitis GJ, Jorcyk CL, Smyth FE and Papas TS. . 1990 Oncogene 5: 1521–1527.

  • Watson DK, McWilliams MJ and Papas TS. . 1988 Virology 164: 99–105.

Download references

Acknowledgements

We thank Xiaohui Xiong and Yevgeniya Suzdaltseva for their excellent technical assistance. This work was supported in part by NIH sponsored multi-institutional program project (CA44768) and SPORE grants (CA58207), an NIH sponsored individual research grant (CA36773), as well as the Hazel P Munroe and Janet Landfear memorial funds. As 1997 – 1998 Visiting Scientist and Professor at the Friedrich Miescher Institute and University of Basel, CC Benz thanks N Hymes, M Burger, and R Herrmann for their assistance throughout that year and acknowledges funding support from the Krebsliga beider Basel.

Author information

Authors and Affiliations

  1. Division of Oncology-Hematology, Department of Medicine, Box-1270, University of California, 3rd and Parnassus, San Francisco, CA 94143-1270, California, USA

    Chuan-Hsiung Chang, Gary K Scott & Christopher C Benz

  2. Department of Neurology, University of California, San Francisco, CA 94143, California, USA

    Michael A Baldwin

  3. Department of Pharmaceutical Chemistry, University of California, San Francisco, CA 94143, California, USA

    Michael A Baldwin

Authors
  1. Chuan-Hsiung Chang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gary K Scott
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Michael A Baldwin
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Christopher C Benz
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chang, CH., Scott, G., Baldwin, M. et al. Exon 4-encoded acidic domain in the epithelium-restricted Ets factor, ESX, confers potent transactivating capacity and binds to TATA-binding protein (TBP). Oncogene 18, 3682–3695 (1999). https://doi.org/10.1038/sj.onc.1202674

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1202674

Keywords

This article is cited by

Search

Advanced search

Quick links