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 Article
  • Published:

Isolation and initial characterization of the BRCA2 promoter

Oncogene volume 18, pages 6000–6012 (1999)Cite this article

Abstract

The hereditary breast cancer susceptibility gene, BRCA2, is considered to be a tumor suppressor gene that may be involved in the cellular response to DNA damage. The transcript for this gene is cell cycle regulated with mRNA levels reaching a peak just before the onset of DNA synthesis. In order to define the mechanisms by which BRCA2 is transcriptionally regulated, we have begun to study upstream regulatory sequences. In this report, we define a minimal promoter region that has strong activity in human breast epithelial cells. Deletions of this sequence narrowed the strong basal activity to a region extending from −66 to +129 with respect to the BRCA2 transcriptional start site. This sequence demonstrated cell cycle regulated activity with kinetics similar to the endogenous transcript. Examination of the sequence revealed several consensus binding sites for transcription factors including an E-box, E2F and Ets recognition motifs. Electrohoretic mobility shift assays revealed specific protein binding to two sequences upstream of the start site; the palindromic E-box and an Ets/E2F site. Site-directed mutagenesis of either of these sites reduced both the basal activity in log phase cells and the cell cycle regulated activity of the promoter. Mutational inactivation of both sites within the same construct effectively eliminated promoter activity. Antibodies to candidate transcription factors used in super shift experiments revealed specific interactions between the BRCA2 promoter and the basic region/helix – loop – helix containing USF-1 and 2 proteins and Elf-1, an Ets domain protein. Binding of these factors depended upon the presence of intact recognition sequences. The USF factors were shown to bind predominantly as a heterodimeric complex of USF-1 and 2 while Elf-1 bound the promoter when it was not occupied by USF. Co-transfection studies with USF proteins and the varicella zoster IE62 protein provide evidence for the involvement of endogenous and exogenous USF in the activation of the BRCA2 promoter. We propose that interactions between USF-1, USF-2 and Elf-1 play an important role in the transcriptional regulation of the BRCA2 gene.

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
Figure 8
Figure 9

Similar content being viewed by others

References

  • Ayer DE and Dynan WS. . 1988 Mol. Cell. Biol. 8: 2021–2033.

  • Band V and Sager R. . 1989 Proc. Natl. Acad. Sci. USA 86: 1249–1253.

  • Bendall AJ and Molloy PL. . 1994 Nucl. Acids Res. 22: 2801–2810.

  • Bertwistle D, Swift S, Marston NJ, Jackson LE, Crossland S, Crompton MR, Marshall CJ and Ashworth A. . 1997 Cancer Res. 57: 5485–5488.

  • Boyd KE and Farnham PJ. . 1997 Mol. Cell. Biol. 17: 2529–2537.

  • Carthew RW, Chodosh LA and Sharp PA. . 1987 Genes Dev. 1: 973–980.

  • Chiang CM and Roeder RG. . 1995 Science 267: 531–536.

  • Cogswell JP, Godlevski MM, Bonham M, Bisi J and Babiss L. . 1995 Mol. Cell. Biol. 15: 2782–2790.

  • Collins N, Wooster R and Stratton MR. . 1997 Br. J. Cancer 76: 1150–1156.

  • Connor F, Bertwistle D, Mee PJ, Ross GM, Swift S, Grigorieva E, Tybulewicz VLJ and Ashworth A. . 1997 Nat. Genet. 17: 423–430.

  • Davis P, Bazar K, Huper G, Lozano G, Marks J and Iglehart JD. . 1996 Oncogene 13: 1315–1322.

  • Desbarats L, Gaubatz S and Eilers M. . 1996 Genes Dev. 10: 447–460.

  • Dobrovic A and Simpfendorfer D. . 1997 Cancer Res. 57: 3347–3350.

  • Du H, Roy AL and Roeder RG. . 1993 EMBO J. 12: 501–511.

  • Ikeda MA, Jakoi L and Nevins JR. . 1996 Proc. Natl. Acad. Sci. USA 93: 3215–3220.

  • Javahery R, Khachi A, Lo K, Zenzie-Gregory B and Smale ST. . 1994 Mol. Cell. Biol. 14: 116–127.

  • Katagiri T, Saito H, Shinohara A, Ogawa H, Kamada N, Nakamura Y and Miki Y. . 1998 Genes Chrom. Cancer 21: 217–222.

  • Krainer M, Silvaarrieta S, Fitzgerald MG, Shimada A, Ishioka C, Kanamaru R, Macdonald DJ, Unsal H, Finkelstein DM, Bowcock A, Isselbacher KJ and Haber DA. . 1997 N. Engl. J. Med. 336: 1416–1421.

  • Lancaster JM, Wooster R, Mangion J, Phelan CM, Cochran C, Gumbs C, Seal S, Barfoot R, Collins N, Bignell G, Patel S, Hamoudi R, Larsson C, Wiseman R, Berchuck A, Iglehart JD, Marks JR, Ashworth A, Stratton M and Futreal PA. . 1996 Nat. Genet. 13: 238–240.

  • Lecine P, Algarte M, Rameil P, Beadling C, Bucher P, Nabholz M and Imbert J. . 1996 Mol. Cell. Biol. 16: 6829–6840.

  • Leiden JM, Wang CY, Petryniak B, Markovitz DM, Nabel GJ and Thompson CB. . 1992 J. Virol. 66: 5890–5897.

  • Li L, Nerlov C, Prendergast G, MacGregor D and Ziff EB. . 1994 EMBO J. 13: 4070–4079.

  • Lim DS and Hasty P. . 1996 Mol. Cell. Biol. 16: 7133–7143.

  • Ludwig T, Chapman DL, Papaioannou VE and Efstratiadis A. . 1997 Genes Dev. 11: 1226–1241.

  • Luo X and Sawadogo M. . 1996 Proc. Natl. Acad. Sci. USA 93: 1308–1313.

  • Mancini DN, Rodenhiser DI, Ainsworth PJ, Omalley FP, Singh SM, Xing WR and Archer TK. . 1998 Oncogene 16: 1161–1169.

  • Marks JR, Huper G, Vaughn JP, Norris J, McDonnel DP, Wiseman RW, Futreal PA and Iglehart JD. . 1997 Oncogene 14: 115–121.

  • Meier JL, Luo X, Sawadogo M and Straus SE. . 1994 Mol. Cell. Biol. 14: 6896–6906.

  • Miltenberger RJ, Sukow KA and Farnham PJ. . 1995 Mol. Cell. Biol. 15: 2527–2535.

  • Miyamoto NG, Moncollin V, Egly JM and Chambon P. . 1985 EMBO J. 4: 3563–3570.

  • Norris J, Fan D, Aleman C, Marks JR, Futreal PA, Wiseman RW, Iglehart JD, Deininger PL and McDonnell DP. . 1995 J. Biol. Chem. 270: 22777–22782.

  • Patel KJ, Yu VPCC, Lee H, Corcoran A, Thistlethwaite FC, Evans MJ, Colledge WH, Friedman LS, Ponder BAJ and Venkitaraman AR. . 1998 Mol. Cell 1: 347–357.

  • Phillips KW, Goldsworthy SM, Bennett LM, Brownlee HA, Wiseman RW and Davis BJ. . 1997 Lab. Invest. 76: 419–425.

  • Qyang Y, Luo X, Lu T, Ismail PM, Krylov D, Vinson C and Sawadogo M. . 1999 Mol. Cell Biol. 19: 1508–1517.

  • Rajan JV, Wang M, Marquis ST and Chodosh LA. . 1996 Proc. Natl. Acad. Sci. USA 93: 13078–13083.

  • Rajan JV, Marquis ST, Gardner HP and Chodosh LA. . 1997 Dev. Biol. 184: 385–401.

  • Rellahan BL, Jensen JP, Howcroft TK, Singer DS, Bonvini E and Weissman AM. . 1998 J. Immunol. 160: 2794–2801.

  • Roeder RG. . 1996 Trends Biochem. Sci. 21: 327–335.

  • Roy AL, Meisterernst M, Pognonec P and Roeder RG. . 1991 Nature 354: 245–248.

  • Sawadogo M and Roeder RG. . 1985 Cell 43: 165–175.

  • Sharan SK, Morimatsu M, Albrecht U, Lim DS, Regel E, Dinh C, Sands A, Eichele G, Hasty P and Bradley A. . 1997 Nature 386: 804–810.

  • Sieweke MH, Tekotte H, Jarosch U and Graf T. . 1998 EMBO J. 17: 1728–1739.

  • Sirito M, Walker S, Lin Q, Kozlowski MT, Klein WH and Sawadogo M. . 1992 Gene Express. 2: 231–240.

  • Sirito M, Lin Q, Maity T and Sawadogo M. . 1994 Nucl. Acids Res. 22: 427–433.

  • Slansky JE and Farnham PJ. . 1996 Curr. Topics Micro. Immunol. 208: 1–30.

  • Smale ST. . 1997 Biochim. Biophys. Acta 1351: 73–88.

  • Smale ST and Baltimore D. . 1989 Cell 57: 103–113.

  • Spillman MA and Bowcock AM. . 1996 Oncogene 13: 1639–1645.

  • Suzuki A, Delapompa JL, Hakem R, Elia A, Yoshida R, Mo R, Nishina H, Chuang T, Wakeham A, Itie A, Koo W, Billia P, Ho A, Fukumoto M, Hui CC and Mak TW. . 1997 Genes Dev. 11: 1242–1252.

  • Tavtigian SV, Simard J, Rommens J, Couch F, Shattuckeidens D, Neuhausen S, Merajver S, Thorlacius S, Offit K, Stoppalyonnet D, Belanger C, Bell R, Berry S, Bogden R, Chen Q, Davis T, Dumont M, Frye C, Hattier T, Jammulapati S, Janecki T, Jiang P, Kehrer R, Leblanc JF, Mitchell JT, McArthur-Morrison J, Nguyen K, Peng Y, Samson C, Schroeder M, Snyder SC, Steele L, Stringfellow M, Stroup C, Swedlund B, Sensen J, Teng D, Thomas A, Tran T, Tranchant M, Weaver-Feldhaus J, Wong AKC, Shizuya H, Eyfjord JE, Cannon-Albright L, Labrie F, Skolnick MH, Weber B, Kamb A and Goldgar DE. . 1996 Nat. Genet. 12: 333–337.

  • Teng DHF, Bogden R, Mitchell J, Baumgard M, Bell R, Berry S, Davis T, Ha PC, Kehrer R, Jammulapati S, Chen QA, Offit K, Skolnick MH, Tavtigian SV, Jhanwar S, Swedlund B, Wong AKC and Kamb A. . 1996 Nat. Genet. 13: 241–244.

  • Thompson CB, Wang CY, Ho IC, Bohjanen PR, Petryniak B, June CH, Miesfeldt S, Zhang L, Nabel GJ, Karpinski B and Leiden JM. . 1992 Mol. Cell. Biol. 12: 1043–1053.

  • Thompson ME, Jensen RA, Obermiller PS, Page DL and Holt JT. . 1995 Nat. Genet. 9: 444–450.

  • Tyler JK and Everett RD. . 1993 Nucl. Acids Res. 21: 513–522.

  • Vaughn JP, Cirisano FD, Huper G, Berchuck A, Futreal PA, Marks JR and Iglehart JD. . 1996a Cancer Res. 56: 4590–4594.

  • Vaughn JP, Davis PL, Jarboe MD, Huper G, Evans AC, Wiseman RW, Berchuck A, Iglehart JD, Futreal PA and Marks JR. . 1996b Cell Growth. Differ. 7: 711–715.

  • Wang CY, Petryniak B, Thompson CB, Kaelin WG and Leiden JM. . 1993 Science 260: 1330–1335.

  • Wong AKC, Pero R, Ormonde PA, Tavtigian SV and Bartel PL. . 1997 J. Biol. Chem. 272.: 31941–31944.

  • Wooster R, Bignell G, Lancaster J, Swift S, Seal S, Mangion J, Collins N, Gregory S, Gumbs C, Micklem G, Barfoot R, Hamoudi R, Patel S, Rice C, Biggs P, Hashim Y, Smith A, Connor F, Arason A, Gudmundsson J, Ficenec D, Kelsell D, Ford D, Tonin P, Bishop DT, Spurr NK, Ponder BAJ, Eeles R, Peto J, Devilee P, Cornelisse C, Lynch H, Narod S, Lenoir G, Egilsson V, Barkadottir RB, Eason DF, Bentley DR, Futreal PA, Ashworth A and Stratton MR. . 1995 Nature 378: 789–792.

  • Zhang HB, Tombline G and Weber BL. . 1998 Cell 92: 433–436.

Download references

Acknowledgements

This work was supported by Public Health Service grants CA-73802 and the Duke Specialized Program of Research Excellence (SPORE) in Breast Cancer CA-68438. The authors would like to thank Gudrun Huper for her excellent technical assistance. We would also like to thank Drs Michelle Sawadogo, Jeffrey Leiden and Stephen Straus for helpful discussions and providing reagents.

Author information

Authors and Affiliations

  1. Department of Surgery, Duke University Medical Center, Box 3873, Durham, 27710, North Carolina, NC, USA

    Penelope L Davis, J Dirk Iglehart & Jeffrey R Marks

  2. The Cancer Genetics Laboratory, Duke University Medical Center, Box 3873, Durham, 27710, North Carolina, NC, USA

    Alexander Miron & Leisha M Andersen

Authors
  1. Penelope L Davis
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Alexander Miron
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Leisha M Andersen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. J Dirk Iglehart
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jeffrey R Marks
    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

Davis, P., Miron, A., Andersen, L. et al. Isolation and initial characterization of the BRCA2 promoter. Oncogene 18, 6000–6012 (1999). https://doi.org/10.1038/sj.onc.1202990

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1202990

Keywords

This article is cited by

Search

Advanced search

Quick links