Abstract
The transforming growth factor-β (TGFβ) type II receptor (TβR-II) is responsible for transducing the growth inhibitory signals of TGFβ. The TβR-II gene promoter lacks both a TATA box and a CAAT box near the transcription initiation site, and has been shown to contain binding sequences for several transcription factors (Sp1, AP1, NF-Y, Cut and ERT) which are important for TβR-II gene promoter activity in vitro. However, it is still not clear which interactions are important for the regulation of TβR-II gene promoter activity in vivo. Using in vivo genomic DNA footprinting of normal human epithelial cells (HaCaT), we have identified two novel identical and strongly protected sites (ggggctgg) at positions −59 and −102 of the TβR-II gene promoter. Mutation of either site significantly reduced promoter activity in transient transfections. Protein binding to these sites, as determined by electrophoretic mobility shift assays (EMSA), was specifically competed with consensus Sp1 oligonucleotides. Furthermore, anti-Sp1/3 antibodies produced band shifts when incubated with the TβR-II −59 and −102 DNA probes. Importantly, Sp1 protein binding was influenced by the presence of an intact NF-Y binding site at position −83. Our data suggests that both Sp1 and NF-Y may play an important role in regulating TβR-II gene promoter basal activity in vivo.
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
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Ammanamanchi S, Brattain MG . 2001 J. Biol. Chem. 276: 3348–3352
Ammanamanchi S, Kim SJ, Sun LZ, Brattain MG . 1998 J. Biol. Chem. 273: 16527–16534
Bae HW, Geiser AG, Kim DH, M.T. C, Burmester JK, Sporn MB, Roberts AB, Kim S-J . 1995 J. Biol. Chem. 270: 29460–29468
Baker JC, Harland RM . 1997 Curr. Opin. Genet. Dev. 7: 467–473
Bennett MK, Osborne TF . 2000 Proc. Natl. Acad. Sci. USA 97: 6340–6344
Birnbaum MJ, van Wijnen AJ, Odgren PR, Last TJ, Suske G, Stein GS, Stein JL . 1995 Biochemistry 34: 16503–16508
Boukamp P, Petrussevska RT, Breitkreutz D, Hornung J, Markham A, Fusenig NE . 1988 J. Cell. Biol. 106: 761–771
Choi S-G, Yi Y, Kim Y-S, Kato M, Chang J, Chung H-W, Hahm KB, Yang H-K, Rhee HH, Kim SJ . 1998 J. Biol. Chem. 273: 110–117
De Luca P, Majello B, Lania L . 1996 J. Biol. Chem. 271: 8533–8536
Doetzlhofer A, Rotheneder H, Lagger G, Koranda M, Kurtev V, Brosch G, Wintersberger E, Seiser C . 1999 Mol. Cell. Biol. 19: 5504–5511
Garrigue-Antar L, Muñoz-Antonia T, Antonia SJ, Gesmonde J, Vellucci VF, Reiss M . 1995 Cancer Res. 55: 3982–3987
Hahm KB, Cho K, Lee C, Im YH, Chang J, Choi SG, Sorensen PH, Thiele CJ, Kim SJ . 1999 Nat. Genet. 23: 222–227
Humphries DE, Bloom BB, Fine A, Goldstein RH . 1994 Biochem. and Biophys. Res. Commun. 203: 1020–1027
Jackson RJ, Antonia SJ, Wright KL, Moon NS, Nepveu A, Muñoz-Antonia T . 1999 Arch. Biochem. Biophys. 371: 290–300
Ji C, Casinghino S, McCarthy TL, Centrella M . 1996 J. Cell. Bichem. 63: 478–490
Kelly D, Kim SJ, Rizzino A . 1998 J. Biol. Chem. 273: 21115–21124
Kennett SB, Udvadia AJ, Horowitz JM . 1997 Nucleic Acids Res. 25: 3110–3117
Kim Y, Ratziu V, Choi SG, Lalazar A, Theiss G, Dang Q, Kim SJ, Friedman SL . 1998 J. Biol. Chem. 273: 33750–33758
Lin HL, Wang XF, Ng-Eaton E, Weinberg RA, Lodish HF . 1992 Cell 68: 775–785
Liu F, Pouponnot C, Massague J . 1997 Genes Dev. 11: 3157–3167
Liu Y, Zhong X, Li W, Brattain MG, Banerji SS . 2000 J. Biol. Chem. 275: 12231–12236
Mantovani R . 1998 Nucleic Acids. Res. 26: 1135–1143
Massague J, Hata A, Liu F . 1997 Trends in Cell. Biol. 7: 187–191
Moustakas A, Kardassis D . 1998 Proc. Natl. Acad. Sci. USA 95: 6733–6738
Muñoz-Antonia T, Li X, Reiss M, Jackson R, Antonia S . 1996 Cancer Res. 56: 4831–4835
Periyasamy S, Ammanamanchi S, Tillekeratne MP, Brattain MG . 2000 Oncogene 19: 4660–4667
Roder K, Wolf SS, Beck KF, Schweizer M . 1997 J. Biol. Chem. 272: 21616–21624
Roder K, Wolf SS, Larkin KJ, Schweizer M . 1999 Gene 234: 61–69
Rotheneder H, Geymayer S, Haidweger E . 1999 J. Mol. Biol. 293: 1005–1015
Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA . 1988 Science 239: 487–491
Sorensen P, Wintersberger E . 1999 J. Biol. Chem. 274: 30943–30949
Sowa Y, Orita T, Hiranabe-Minamikawa S, Nakano K, Mizuno T, Nomura H, Sakai T . 1999 Ann. N Y Acad. Sci. 886: 195–199
Steffen ML, Harrison WR, Elder FF, Cook GA, Park EA . 1999 Biochem. J. 340: 425–432
Sun L, Wu G, Willson JKV, Zborowski E, Yang J, Rajkarunanayake I, Wang J, Gentry LE, Wang X-L, Brattain M . 1994 J. Biol. Chem. 269: 26449–26455
Suske G . 1999 Gene 238: 291–300
Wang J, Sun L, Myeroff L, Wang X, Gentry LE, Yang J, Liang J, Zborowska E, Markowitz S, Willson JKV, Brattain MG . 1995 J. Biol. Chem. 270: 22044–22049
Wrana J, Attisano L, Weiser R, Ventura F, Massaque J . 1994 Nature 370: 341–347
Wrana JL, Attisano L, Carcamo J, Zentella A, Doody J, Laiho M, Wang X-F, Massaque J . 1992 Cell 71: 1003–1014
Wright KL, Moore TL, Vilen BJ, Brown AM, Ting JP . 1995 J. Biol. Chem. 270: 20978–20986
Acknowledgements
We would like to thank Dr Nancy Olashaw for critically reading this manuscript. This study was partially supported by the American Cancer Society Grant #RPG-98-039-01-CNE.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Jennings, R., Alsarraj, M., Wright, K. et al. Regulation of the human transforming growth factor β type II receptor gene promoter by novel Sp1 sites. Oncogene 20, 6899–6909 (2001). https://doi.org/10.1038/sj.onc.1204808
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.onc.1204808
Keywords
This article is cited by
-
Modulation of transforming growth factor beta signalling pathway genes by transforming growth factor beta in human osteoarthritic chondrocytes: involvement of Sp1 in both early and late response cells to transforming growth factor beta
Arthritis Research & Therapy (2011)
-
Therapeutic effects of the Sp1 inhibitor mithramycin A in glioblastoma
Journal of Neuro-Oncology (2011)