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:

Transcriptional Control and Deregulation

Transcriptional regulation of parathyroid hormone-related protein promoter P2 by NF-κB in adult T-cell leukemia/lymphoma

Leukemia volume 21, pages 1752–1762 (2007)Cite this article

Abstract

Parathyroid hormone-related protein (PTHrP) plays a primary role in the development of humoral hypercalcemia of malignancy (HHM) that occurs in the majority of patients with adult T-cell leukemia/lymphoma (ATLL) due to human T-cell lymphotropic virus type-1 (HTLV-1) infection. We previously showed that ATLL cells constitutively express high levels of PTHrP via activation of promoters P2 and P3, resulting in HHM. In this study, we characterized a nuclear factor-κB (NF-κB) binding site in the P2 promoter of human PTHrP. Using electrophoretic mobility shift assays, we detected a specific complex in Tax-expressing human T cells composed of p50/c-Rel, and two distinct complexes in ATLL cells consisting of p50/p50 homodimers and a second unidentified protein(s). Chromatin immunoprecipitation assays confirmed in vivo binding of p50 and c-Rel on the PTHrP P2 promoter. Using transient co-transfection with NF-κB expression plasmids and PTHrP P2 luciferase reporter-plasmid, we showed that NF-κB p50/p50 alone and p50/c-Rel or p50/Bcl-3 cooperatively upregulated the PTHrP P2 promoter. Furthermore, inhibition of NF-κB activity by Bay 11-7082 reduced PTHrP P2 promoter-initiated transcripts in HTLV-1-infected T cells. In summary, the data demonstrated that transcriptional regulation of PTHrP in ATLL cells can be controlled by NF-κB activation and also suggest a Tax-independent mechanism of activation of PTHrP in ATLL.

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

Similar content being viewed by others

Accession codes

Accessions

GenBank/EMBL/DDBJ

References

  1. Poiesz BJ, Ruscetti FW, Gazdar AF, Bunn PA, Minna JD, Gallo RC . Detection and isolation of type C retrovirus particles from fresh and cultured lymphocytes of a patient with cutaneous T-cell lymphoma. Proc Natl Acad Sci USA 1980; 77: 7415–7419.

    Article  CAS  Google Scholar 

  2. Broadus AE, Mangin M, Ikeda K, Insogna KL, Weir EC, Burtis WJ et al. Humoral hypercalcemia of cancer. Identification of a novel parathyroid hormone-like peptide. N Engl J Med 1988; 319: 556–563.

    Article  CAS  Google Scholar 

  3. Grill V, Martin TJ . Hypercalcemia of malignancy. Rev Endocr Metab Disord 2000; 1: 253–263.

    Article  CAS  Google Scholar 

  4. Wysolmerski JJ, Stewart AF . The physiology of parathyroid hormone-related protein: an emerging role as a developmental factor. Annu Rev Physiol 1998; 60: 431–460.

    Article  CAS  Google Scholar 

  5. Mangin M, Ikeda K, Dreyer BE, Broadus AE . Identification of an up-stream promoter of the human parathyroid hormone-related peptide gene. Mol Endocrinol 1990; 4: 851–858.

    Article  CAS  Google Scholar 

  6. Campos RV, Wang C, Drucker DJ . Regulation of parathyroid hormone-related peptide (PTHrP) gene transcription. Mol Endocrinol 1992; 6: 1642–1652.

    CAS  PubMed  Google Scholar 

  7. Suva LJ, Mather KA, Gillespie MT, Webb GC, Ng KW, Winslow GA et al. Structure of the 5′ flanking region of the gene encoding human parathyroid-hormone-related protein (PTHrP). Gene 1989; 77: 95–105.

    Article  CAS  Google Scholar 

  8. Vasavada RC, Wysolmerski JJ, Broadus AE, Philbrick WM . Identification and characterization of a GC-rich promoter of the human parathyroid hormone-related peptide gene. Mol Endocrinol 1993; 7: 273–282.

    CAS  PubMed  Google Scholar 

  9. Bouizar Z, Spyratos F, De Vernejoul MC . The parathyroid hormone-related protein (PTHrP) gene: use of downstream TATA promotor and PTHrP 1–139 coding pathways in primary breast cancers vary with the occurrence of bone metastasis. J Bone Miner Res 1999; 14: 406–414.

    Article  CAS  Google Scholar 

  10. Southby J, Murphy LM, Martin TJ, Gillespie MT . Cell-specific and regulator-induced promoter usage and messenger ribonucleic acid splicing for parathyroid hormone-related protein. Endocrinology 1996; 137: 1349–1357.

    Article  CAS  Google Scholar 

  11. Richard V, Luchin A, Brena RM, Plass C, Rosol TJ . Quantitative evaluation of alternative promoter usage and 3′ splice variants for parathyroid hormone-related protein by real-time reverse transcription-PCR. Clin Chem 2003; 49: 1398–1402.

    Article  CAS  Google Scholar 

  12. Brandt DW, Bruns ME, Bruns DE, Ferguson JE, Burton DW, Deftos LJ . The parathyroid hormone-related protein (PTHrP) gene preferentially utilizes a GC-rich promoter and the PTHrP 1–139 coding pathway in normal human amnion. Biochem Biophys Res Commun 1992; 189: 938–943.

    Article  CAS  Google Scholar 

  13. Good L, Maggirwar SB, Sun SC . Activation of the IL-2 gene promoter by HTLV-I tax involves induction of NF-AT complexes bound to the CD28-responsive element. EMBO J 1996; 15: 3744–3750.

    Article  CAS  Google Scholar 

  14. Ballard DW, Bohnlein E, Lowenthal JW, Wano Y, Franza BR, Greene WC . HTLV-I tax induces cellular proteins that activate the kappa B element in the IL-2 receptor alpha gene. Science 1988; 241: 1652–1655.

    Article  CAS  Google Scholar 

  15. Duyao MP, Kessler DJ, Spicer DB, Sonenshein GE . Transactivation of the c-myc gene by HTLV-1 tax is mediated by NFkB. Curr Top Microbiol Immunol 1992; 182: 421–424.

    CAS  PubMed  Google Scholar 

  16. Mori N, Fujii M, Ikeda S, Yamada Y, Tomonaga M, Ballard DW et al. Constitutive activation of NF-kappaB in primary adult T-cell leukemia cells. Blood 1999; 93: 2360–2368.

    CAS  PubMed  Google Scholar 

  17. Hironaka N, Mochida K, Mori N, Maeda M, Yamamoto N, Yamaoka S . Tax-independent constitutive IkappaB kinase activation in adult T-cell leukemia cells. Neoplasia 2004; 6: 266–278.

    Article  CAS  Google Scholar 

  18. May MJ, Ghosh S . Signal transduction through NF-kappa B. Immunol Today 1998; 19: 80–88.

    Article  CAS  Google Scholar 

  19. Leung TH, Hoffmann A, Baltimore D . One nucleotide in a kappaB site can determine cofactor specificity for NF-kappaB dimers. Cell 2004; 118: 453–464.

    Article  CAS  Google Scholar 

  20. Hayden MS, Ghosh S . Signaling to NF-kappaB. Genes Dev 2004; 18: 2195–2224.

    Article  CAS  Google Scholar 

  21. Bours V, Franzoso G, Azarenko V, Park S, Kanno T, Brown K et al. The oncoprotein Bcl-3 directly transactivates through kappa B motifs via association with DNA-binding p50B homodimers. Cell 1993; 72: 729–739.

    Article  CAS  Google Scholar 

  22. Franzoso G, Bours V, Park S, Tomita-Yamaguchi M, Kelly K, Siebenlist U . The candidate oncoprotein Bcl-3 is an antagonist of p50/NF-kappa B-mediated inhibition. Nature 1992; 359: 339–342.

    Article  CAS  Google Scholar 

  23. Bundy DL, McKeithan TW . Diverse effects of BCL3 phosphorylation on its modulation of NF-kappaB p52 homodimer binding to DNA. J Biol Chem 1997; 272: 33132–33139.

    Article  CAS  Google Scholar 

  24. Caamano JH, Perez P, Lira SA, Bravo R . Constitutive expression of Bc1-3 in thymocytes increases the DNA binding of NF-kappaB1 (p50) homodimers in vivo. Mol Cell Biol 1996; 16: 1342–1348.

    Article  CAS  Google Scholar 

  25. Richard V, Nadella MV, Green PL, Lairmore MD, Feuer G, Foley JG et al. Transcriptional regulation of parathyroid hormone-related protein promoter P3 by ETS-1 in adult T-cell leukemia/lymphoma. Leukemia 2005; 19: 1175–1183.

    Article  CAS  Google Scholar 

  26. Feuer G, Zack JA, Harrington Jr WJ, Valderama R, Rosenblatt JD, Wachsman W et al. Establishment of human T-cell leukemia virus type I T-cell lymphomas in severe combined immunodeficient mice. Blood 1993; 82: 722–731.

    CAS  PubMed  Google Scholar 

  27. Phillips KE, Herring B, Wilson LA, Rickford MS, Zhang M, Goldman CK et al. IL-2Ralpha-Directed monoclonal antibodies provide effective therapy in a murine model of adult T-cell leukemia by a mechanism other than blockade of IL-2/IL-2Ralpha interaction. Cancer Res 2000; 60: 6977–6984.

    CAS  PubMed  Google Scholar 

  28. Stewart SA, Feuer G, Jewett A, Lee FV, Bonavida B, Chen IS . HTLV-1 gene expression in adult T-cell leukemia cells elicits an NK cell response in vitro and correlates with cell rejection in SCID mice. Virology 1996; 226: 167–175.

    Article  CAS  Google Scholar 

  29. Mori N, Gill PS, Mougdil T, Murakami S, Eto S, Prager D . Interleukin-10 gene expression in adult T-cell leukemia. Blood 1996; 88: 1035–1045.

    CAS  PubMed  Google Scholar 

  30. Guttridge DC, Albanese C, Reuther JY, Pestell RG, Baldwin Jr AS . NF-kappaB controls cell growth and differentiation through transcriptional regulation of cyclin D1. Mol Cell Biol 1999; 19: 5785–5799.

    Article  CAS  Google Scholar 

  31. Cheng AS, Jin VX, Fan M, Smith LT, Liyanarachchi S, Yan PS et al. Combinatorial analysis of transcription factor partners reveals recruitment of c-MYC to estrogen receptor-alpha responsive promoters. Mol Cell 2006; 21: 393–404.

    Article  CAS  Google Scholar 

  32. Dirksen WP, Mohamed SA, Fisher SA . Splicing of a myosin phosphatase targeting subunit 1 alternative exon is regulated by intronic cis-elements and a novel bipartite exonic enhancer/silencer element. J Biol Chem 2003; 278: 9722–9732.

    Article  CAS  Google Scholar 

  33. Nishikori M, Maesako Y, Ueda C, Kurata M, Uchiyama T, Ohno H . High-level expression of BCL3 differentiates t(2;5)(p23;q35)-positive anaplastic large cell lymphoma from Hodgkin disease. Blood 2003; 101: 2789–2796.

    Article  CAS  Google Scholar 

  34. Holz LE, Jakobsen KP, Van Snick J, Cormont F, Sewell WA . Dexamethasone inhibits IL-9 production by human T cells. J Inflamm (Lond) 2005; 2: 3.

    Article  Google Scholar 

  35. Glasgow JN, Wood T, Perez-Polo JR . Identification and characterization of nuclear factor kappaB binding sites in the murine bcl-x promoter. J Neurochem 2000; 75: 1377–1389.

    Article  CAS  Google Scholar 

  36. Jeang KT, Derse D, Matocha M, Sharma O . Expression status of Tax protein in human T-cell leukemia virus type 1-transformed MT4 cells: recall of MT4 cells distributed by the NIH AIDS Research and Reference Reagent Program. J Virol 1997; 71: 6277–6278.

    CAS  PubMed  PubMed Central  Google Scholar 

  37. Richard V, Lairmore MD, Green PL, Feuer G, Erbe RS, Albrecht B et al. Humoral hypercalcemia of malignancy: severe combined immunodeficient/beige mouse model of adult T-cell lymphoma independent of human T-cell lymphotropic virus type-1 tax expression. Am J Pathol 2001; 158: 2219–2228.

    Article  CAS  Google Scholar 

  38. Richard V, Luchin A, Brena RM, Plass C, Rosol TJ . Quantitative evaluation of alternative promoter usage and 3′ splice variants for parathyroid hormone-related protein by real-time reverse transcription-PCR. Clin Chem 2003; 49: 1398–1402.

    Article  CAS  Google Scholar 

  39. Vasavada RC, Wysolmerski JJ, Broadus AE, Philbrick WM . Identification and characterization of a GC-rich promoter of the human parathyroid hormone-related peptide gene. Mol Endocrinol 1993; 7: 273–282.

    CAS  PubMed  Google Scholar 

  40. Peloponese JM, Yeung ML, Jeang KT . Modulation of nuclear factor-kappaB by human T cell leukemia virus type 1 Tax protein: implications for oncogenesis and inflammation. Immunol Res 2006; 34: 1–12.

    Article  CAS  Google Scholar 

  41. Sun SC, Yamaoka S . Activation of NF-kappaB by HTLV-I and implications for cell transformation. Oncogene 2005; 24: 5952–5964.

    Article  CAS  Google Scholar 

  42. Dittmer J, Pise-Masison CA, Clemens KE, Choi KS, Brady JN . Interaction of human T-cell lymphotropic virus type I Tax, Ets1, and Sp1 in transactivation of the PTHrP P2 promoter. J Biol Chem 1997; 272: 4953–4958.

    Article  CAS  Google Scholar 

  43. Mori N, Ejima E, Prager D . Transactivation of parathyroid hormone-related protein gene expression by human T-cell leukemia virus type I tax. Eur J Haematol 1996; 56: 116–117.

    Article  CAS  Google Scholar 

  44. Ikeda K, Inoue D, Okazaki R, Kikuchi T, Ogata E, Matsumoto T . Parathyroid hormone-related peptide in hypercalcemia associated with adult T cell leukemia/lymphoma: molecular and cellular mechanism of parathyroid hormone-related peptide overexpression in HTLV-I-infected T cells. Miner Electrolyte Metab 1995; 21: 166–170.

    CAS  PubMed  Google Scholar 

  45. Richard V, Nadella MV, Green PL, Lairmore MD, Feuer G, Foley JG et al. Transcriptional regulation of parathyroid hormone-related protein promoter P3 by ETS-1 in adult T-cell leukemia/lymphoma. Leukemia 2005; 19: 1175–1183.

    Article  CAS  Google Scholar 

  46. Li CC, Ruscetti FW, Rice NR, Chen E, Yang NS, Mikovits J et al. Differential expression of Rel family members in human T-cell leukemia virus type I-infected cells: transcriptional activation of c-rel by Tax protein. J Virol 1993; 67: 4205–4213.

    CAS  PubMed  PubMed Central  Google Scholar 

  47. Kurland JF, Kodym R, Story MD, Spurgers KB, McDonnell TJ, Meyn RE . NF-kappaB1 (p50) homodimers contribute to transcription of the bcl-2 oncogene. J Biol Chem 2001; 276: 45380–45386.

    Article  CAS  Google Scholar 

  48. Fujita T, Nolan GP, Liou HC, Scott ML, Baltimore D . The candidate proto-oncogene bcl-3 encodes a transcriptional coactivator that activates through NF-kappa B p50 homodimers. Genes Dev 1993; 7: 1354–1363.

    Article  CAS  Google Scholar 

  49. Lin R, Gewert D, Hiscott J . Differential transcriptional activation in vitro by NF-kappa B/Rel proteins. J Biol Chem 1995; 270: 3123–3131.

    Article  CAS  Google Scholar 

  50. Schreiber J, Jenner RG, Murray HL, Gerber GK, Gifford DK, Young RA . Coordinated binding of NF-kappaB family members in the response of human cells to lipopolysaccharide. Proc Natl Acad Sci USA 2006; 103: 5899–5904.

    Article  CAS  Google Scholar 

  51. Bundy DL, McKeithan TW . Diverse effects of BCL3 phosphorylation on its modulation of NF-kappaB p52 homodimer binding to DNA. J Biol Chem 1997; 272: 33132–33139.

    Article  CAS  Google Scholar 

  52. Watanabe N, Iwamura T, Shinoda T, Fujita T . Regulation of NFKB1 proteins by the candidate oncoprotein BCL-3: generation of NF-kappaB homodimers from the cytoplasmic pool of p50-p105 and nuclear translocation. EMBO J 1997; 16: 3609–3620.

    Article  CAS  Google Scholar 

  53. Mori N, Yamada Y, Ikeda S, Yamasaki Y, Tsukasaki K, Tanaka Y et al. Bay 11-7082 inhibits transcription factor NF-kappaB and induces apoptosis of HTLV-I-infected T-cell lines and primary adult T-cell leukemia cells. Blood 2002; 100: 1828–1834.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the National Cancer Institute (CA100730 and CA77911). MVPN was supported by the Glenn C Barber Fellowship from the College of Veterinary Medicine, The Ohio State University; TR and SS were supported by the National Center for Research Resources (RR00168) and the NCRR T32 (RR07073), respectively.

Author information

Authors and Affiliations

  1. Department of Veterinary Biosciences, The Ohio State University, Columbus, OH, USA

    M V P Nadella, W P Dirksen, S Shu, J A Morgenstern & T J Rosol

  2. Human Cancer Genetics, The Ohio State University, Columbus, OH, USA

    K S Nadella, A S Cheng, T H Huang & D Guttridge

  3. Pfizer, Sandwich Laboratories, Kent, UK

    V Richard

  4. Center for Biostatistics, The Ohio State University, Columbus, OH, USA

    S A Fernandez

  5. Center for Retrovirus Research, The Ohio State University, Columbus, OH, USA

    T J Rosol

Authors
  1. M V P Nadella
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. W P Dirksen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. K S Nadella
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. S Shu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. A S Cheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. J A Morgenstern
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. V Richard
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. S A Fernandez
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. T H Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. D Guttridge
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. T J Rosol
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to T J Rosol.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nadella, M., Dirksen, W., Nadella, K. et al. Transcriptional regulation of parathyroid hormone-related protein promoter P2 by NF-κB in adult T-cell leukemia/lymphoma. Leukemia 21, 1752–1762 (2007). https://doi.org/10.1038/sj.leu.2404798

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.leu.2404798

Keywords

This article is cited by

Search

Advanced search

Quick links