Abstract
In the present study, we report that, despite the presence of one perfect p53 consensus sequence homology (designated SCL CS) and four half-sites within the 3′-untranslated region of the stem cell leukemia (SCL) gene, the native endogenous gene is not regulated by p53. We employ a tet-repressible system to show that, under conditions in which the WAF1 mRNA steady-state level is upregulated fourfold by p53, the SCL mRNA level is not altered. In a previous report, we demonstrated that p53 interactions with the SCL CS can upregulate downstream reporter gene activity 43-fold in transient reporter assays. This disparity prompted us to explore the differences between p53 regulation of SCL CS activity in organized (chromosomally integrated) and disorganized (non-replicating episomal plasmid) chromatin. We show that p53 can increase (between 3 – 80-fold), decrease (between 5 – 33-fold) or have no effect upon transactivation of an SCL CS/reporter fusion gene depending upon chromosomal integration site. Most studies used to characterize p53 binding sites employ transient transfection assays. Our results suggest that characterization of consensus sequence homologies by assay of transiently transfected cells may be inaccurate.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Ackland-Berglund C and Leib DA. . 1995 BioTechniques 18: 196–200.
Begley CG, Aplan PD, Denning SM, Hayes BF, Waldmann TA and Kirsch IR. . 1989 Proc. Natl. Acad. Sci. USA 86: 10128–10132.
Bi S, Lanza F and Goldman JM. . 1994 Cancer Res. 54: 582–586.
Chin PL, Momand J and Pfeifer GP. . 1997 Oncogene 15: 87–99.
Chomczynski P and Sacchi N. . 1987 Anal. Biochem. 162: 156–159.
Cook JL, Re RN, Giardina JF, Fontenot FE, Cheng DY and Alam J. . 1995 Oncogene 11: 723–733.
Deb SP, Munoz RM, Brown DR, Subler MA and Deb S. . 1994 Oncogene 9: 1341–1349.
El-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, Lin D, Mercer WE, Kinzler KW and Vogelstein B. . 1993 Cell 75: 817–825.
Farmer G, Bargonetti J, Zhu H, Friedman P, Prywes R and Prives C. . 1992 Nature 358: 83–85.
Funk WD, Pak DT, Karas RH, Wright WE and Shay JW. . 1992 Mol. Cell. Biol. 12: 2866–2871.
Furth PA, St. Onge L, Boger H, Gruss P, Gossen M, Kistner A, Bujard H and Henninghausen L. . 1994 Proc. Natl. Acad. Sci. USA 91: 9302–9306.
Gossen M and Bujard H. . 1992 Proc. Natl. Acad. Sci. USA 89: 5547–5551.
Howe JR, Skryabin BV, Belcher SM, Zerillo CA and Schmauss C. . 1995 J. Biol. Chem. 270: 14168–14174.
Jackson P and Yardley G. . 1997 FEBS Lett. 406: 271–274.
Jackson P, Mastrangelo I, Reed M, Tegtmeyer P, Yardley G and Barrett J. . 1998 Oncogene 16: 283–292.
Kristjuhand A and Maimets T. . 1995 Eur. J. Biochem. 234: 827–831.
Mack DH, Vartikar J, Pipas JM and Laimins LA. . 1993 Nature 363: 281–283.
Metcalfe AMJ, Dixon RM and Radda GK. . 1997 Nucleic Acids Res. 25: 983–986.
Morris GF, Bischoff JR and Matthews MB. . 1996 Proc. Natl. Acad. Sci. USA 93: 895–899.
O'Connor DJ, Lam EW, Griffin S, Zhong S, Leighton LC, Burbidge SA and Lu X. . 1995 EMBO J. 14: 6184–6192.
Osifchin NE, Jiang D, Ohtani-Fujita N, Fujita T, Carroza M, Kim S-J, Saki T and Robbins PD. . 1994 J. Biol. Chem. 269: 6383–6389.
Park DJ, Nakamura H, Chumakov AM, Said JW, Miller CW, Chen DL and Koeffler HP. . 1994 Oncogene 9: 1899–1906.
Pennie WD, Hager GL and Smith CL. . 1995 Mol. Cell. Biol. 15: 2125–2134.
Ponte P, Ng SY, Engel J, Gunning P and Kedes L. . 1984 Nucleic Acids Res. 3: 1687–1696.
Raimond J, Rouleux F, Monsigny M and Legrand A. . 1995 FEBS Lett. 363: 165–169.
Raycroft L, Schmidt JR, Yoas K, Hao M and Lozano G. . 1991 Mol. Cell. Biol. 11: 6067–6074.
Santhanam U, Ray A and Sehgal PB. . 1991 Proc. Natl. Acad. Sci. USA 88: 7605–7609.
Zauberman A, Flusberg D, Haupt Y, Barak Y and Oren M. . 1995 Nucleic Acids Res. 23: 2584–2592.
Acknowledgements
This work was supported by the Alton Ochsner Medical Foundation.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Cook, J., Zhang, Z., Alam, J. et al. Effects of chromosomal integration site upon p53 interactions with DNA consensus sequence homologies. Oncogene 18, 2373–2379 (1999). https://doi.org/10.1038/sj.onc.1202566
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.onc.1202566