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Horm Metab Res 2002; 34(6): 303-310
DOI: 10.1055/s-2002-33258
Original Basic
© Georg Thieme Verlag Stuttgart · New York

APE/Ref-1 is Controlled by Both Redox and cAMP-Dependent Mechanisms in Rat Thyroid Cells

G.  Tell 2 , A.  Pines 2 , M.  Pandolfi 3 , A.  V.  D’Elia 1 , D.  Donnini 1 , R.  Lonigro 1 , G.  Manzini 2 , D.  Russo 4 , C.  Di Loreto 3 , G.  Damante 1
  • 1Dipartimento di Scienze e Tecnologie Biomediche, Università di Udine. M.A.T.I. Center.
  • 2Dipartimento di Biochimica Biofisica e Chimica delle Macromolecole, Università di Trieste, C.E.B.
  • 3Dipartimento di Ricerche Mediche e Morfologiche, Università di Udine.
  • 4Dipartimento di Scienze Farmacobiologiche, Università di Catanzaro
Further Information

Publication History

Received 26 July 2001

Accepted after Revision 12 February 2002

Publication Date:
12 August 2002 (online)

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Abstract

APE/Ref-1 is a multifunctional protein possessing both redox and DNA repair functions. Through its redox activity, APE/Ref-1 controls the DNA-binding function of several transcriptional regulators (AP1, NF-κB, p53, Pax proteins). We have previously shown that APE/Ref-1 upregulates the transcriptional activity of the thyroid-specific transcription factor Pax8. In thyroid cells, APE/Ref-1 can be detected both in the nuclear and cytoplasmatic compartments. In this study regulatory mechanisms acting on APE/Ref-1 were revealed using the FRTL-5 cell line. TSH induces both cytoplasm-to-nucleus translocation and neosynthesis of APE/Ref-1 protein. Interestingly, only neosynthesis is dependent on cAMP signalling. In contrast, the cytoplasm-to-nucleus translocation is dependent on redox-mediated mechanisms. Based upon the data shown in this study and in others, a bimodal control of APE/Ref-1 by TSH can be delineated.

Key words

APE/Ref-1 - Redox Potential - Nuclear Translocation - Thyroid - cAMP

References

  • 1 Nakamura H, Nakamura K, Yodoi J. Redox regulation of cellular activation.  Annu Rev Immunol. 1997;  15 351-369
    CrossrefPubMedGoogle Scholar
  • 2 Rothwell D G, Barzilay G, Gorman M, Morera S, Freemont P, Hickson I D. The structure and functions of the HAP1/Ref-1 protein.  Oncol Res. 1997;  9 275-280
    PubMedGoogle Scholar
  • 3 Ramana C V, Boldogh I, Izumi T, Mitra S. Activation of apurinic/apyrimidinic endonuclease in human cells by reactive oxygen species and its correlation with their adaptive response to genotoxicity of free radicals.  Proc Natl Acad Sci USA. 1998;  95 5061-5066
    CrossrefPubMedGoogle Scholar
  • 4 Xanthoudakis S, Smeyne R J, Wallace J D, Curran T. The redox/DNA repair protein, Ref-1, is essential for early embryonic development in mice.  Proc Natl Acad Sci USA. 1996;  93 8919-8923
    CrossrefPubMedGoogle Scholar
  • 5 Gaiddon C, Moorthy N C, Prives C. Ref-1 regulates the transactivation and pro-apoptotic functions of p53 in vivo.  EMBO J. 1999;  18 5609-5621
    CrossrefPubMedGoogle Scholar
  • 6 Grösh S, Fritz G, Kaina B. Apurinic endonuclease (Ref-1) is induced in mammalian cells by oxidative stress and involved in clastogenic adaptation.  Cancer Res. 1998;  58 4410-4416
    PubMedGoogle Scholar
  • 7 Tell G, Zecca A, Pellizzari L, Spessotto P, Colombatti A, Kelley M R, Damante G, Pucillo C. An “environment to nucleus” signaling system operates in B lymphocytes: redox status modulates BSAP/Pax-5 activation through Ref-1 nuclear translocation.  Nucl Acid Res. 2000;  28 1099-1105
    PubMedGoogle Scholar
  • 8 Raspe E, Dumont J E. Tonic modulation of dog thyrocyte H2O2 generation and I-uptake by thyrotropin through the cyclic adenosine 3’, 5’-monophosphate cascade.  Endocrinology. 1995;  136 965-973
    CrossrefPubMedGoogle Scholar
  • 9 Bianchi G, Solaroli E, Zaccheroni V, Grossi G, Bargossi A M, Melchionda N, Marchesini G. Oxidative stress and anti-oxidant metabolites in patients with hyperthyroidism: effect of treatment. Horm Metab.  Res. 1999;  31 620-624
    PubMedGoogle Scholar
  • 10 Tell G, Pellizzari L, Pucillo C, Puglisi F, Cesselli D, Kelley M R, di Loreto C, Damante G. TSH controls Ref-1 nuclear translocation in thyroid cells. J Mol.  Endocrinol. 2000;  24 383-390
    PubMedGoogle Scholar
  • 11 Kakolyris S, Kaklamanis L, Giatromanolaki A, Koukourakis M, Hickson I D, Barzilay G, Turley H, Leek R D, Kanavaros P, Georgulias V, Gatter K C, Harris A L. Expression and subcellular localization of human AP endonuclease 1 (HAP1/Ref-1) protein: a basis for its role in human disease.  Histopathology. 1998;  33 561-569
    CrossrefPubMedGoogle Scholar
  • 12 Tell G, Pellizzari L, Cimarosti D, Pucillo C, Damante G. Ref-1 controls pax-8 DNA-binding activity.  Biochem Biophys Res Commun. 1998;  252 178-183
    CrossrefPubMedGoogle Scholar
  • 13 Ambesi-Imbiombato F S, Coon H G. Thyroid cells in culture.  Int Rev Cytol. 1979;  10 163-172
    PubMedGoogle Scholar
  • 14 Tell G, Scaloni A, Pellizzari L, Formisano S, Pucillo C, Damante G. Redox potential controls the structure and DNA binding activity of the paired domain.  J Biol Chem. 1998;  273 25 062-25 072
    PubMedGoogle Scholar
  • 15 Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.  Anal Biochem. 1976;  72 248-254
    CrossrefPubMedGoogle Scholar
  • 16 Xu Y, Moore D, Broshears J, Liu L, Wilson T M, Kelley M R. The apurinic/apyrimidinic endonuclease (APE/ref-1) DNA repair enzyme is elevated in premalignant and malignant cervical cancer.  Anticancer Res. 1997;  37 13-20
    PubMedGoogle Scholar
  • 17 Moore D H, Michael H, Tritt R, Parsons S H, Kelley M R. Alterations in the expression of the DNA repair/redox enzyme APE/ref-1 in epithelial ovarian cancers.  Clin Cancer Res. 2000;  6 602-609
    PubMedGoogle Scholar
  • 18 di Monte D, Ross D, Bellomo G, Eklow L, Orrenius S. Alterations in intracellular thiol homeostasis during the metabolism of menadione by isolated rat hepatocytes.  Arch Biochem Biophys. 1984;  235 334-342
    CrossrefPubMedGoogle Scholar
  • 19 Dumont J E, Lamy F, Roger P, Maenhaut C. Physiological and pathological regulation of thyroid cell proliferation and differentiation by thyrotropin and other factors.  Physiol Rev. 1992;  72 667-696
    CrossrefPubMedGoogle Scholar
  • 20 Asai T, Kambe F, Kikumori T, Seo H. Increase in Ref-1 mRNA and protein by thyrotropin in rat thyroid FRTL-5 cells.  Biochem Biophys Res Commun. 1997;  236 71-74
    CrossrefPubMedGoogle Scholar
  • 21 Nunez J, Pommier J. Formation of thyroid hormones.  Vitam Horm. 1982;  39 175-229
    PubMedGoogle Scholar
  • 22 Bjorkman U, Ekholm R. Accelerated exocytosis and H2O2 generation in isolated thyroid follicles enhance protein iodination.  Endocrinology. 1988;  130 393-399
    PubMedGoogle Scholar
  • 23 Sies H. Glutathione and its role in cellular functions.  Free Radic Biol Med. 1999;  27 916-921
    CrossrefPubMedGoogle Scholar
  • 24 Lonigro R, Donnini D, Fabbro D, Perrell G, Damante G, Ambesi-Impiombato F S, Curcio F. Thyroid-specific gene expression is differentially influenced by intracellular glutathione level in FRTL-5 cells.  Endocrinology. 2000;  141 901-909
    CrossrefPubMedGoogle Scholar
  • 25 Saji M, Ikuyama S, Akamizu T, Kohn L D. Increases in cytosolic Ca++ down regulate thyrotropin receptor gene expression by a mechanism different from the cAMP signal.  Biochem Biophys Res Commun. 1997;  176 94-101
    PubMedGoogle Scholar
  • 26 Bizzarri C, Corda D. Norepinephrine, unlike ATP, induces all-or-none increase in cytosolic calcium in thyroid cells. The role of inositol-trisphosphate-sensitive stores and calcium channels. Eur J.  Biochem. 1994;  219 837-844
    PubMedGoogle Scholar
  • 27 Corda D, Marcocci C, Kohn L D, Axelrod J, Luini A. Association of the changes in cytosolic Ca2+ and iodide efflux induced by thyrotropin and by the stimulation of alpha 1-adrenergic receptors in cultured rat thyroid cells.  J Biol Chem. 1985;  260 9230-9236
    PubMedGoogle Scholar
  • 28 Grösh S, Kaina B. Transcriptional activation of apurinic/apyrimidinic endonuclease (Ape, Ref-1) by oxidative stress requires CREB.  Biochem Biophys Res Commun. 1999;  261 859-863
    CrossrefPubMedGoogle Scholar
  • 29 Raspe E, Laurent E, Corvilain B, Verjans B, Erneux C, Dumont J E. Control of the intracellular Ca(2+)-concentration and the inositol phosphate accumulation in dog thyrocyte primary culture: evidence for different kinetics of Ca(2+)-phosphatidylinositol cascade activation and for involvement in the regulation of H2O2 production.  J Cell Physiol. 1991;  146 242-250
    CrossrefPubMedGoogle Scholar
  • 30 Corvilain B, Laurent E, Lecomte M, Vansande J, Dumont J E. Role of the cyclic adenosine 3’,5’-monophosphate and the phosphatidylinositol-Ca2+ cascades in mediating the effects of thyrotropin and iodide on hormone synthesis and secretion in human thyroid slices.  J Clin Endocrinol Metab. 1994;  79 152-159
    CrossrefPubMedGoogle Scholar
  • 31 Kimura T, Okajima F, Sho K, Kobayashi I, Kondo Y. Thyrotropin-induced hydrogen peroxide production in FRTL-5 thyroid cells is mediated not by adenosine 3’,5’-monophosphate, but by Ca2+ signaling followed by phospholipase-A2 activation and potentiated by an adenosine derivative.  Endocrinology. 1995;  136 116-123
    CrossrefPubMedGoogle Scholar
  • 32 Okazaki T, Chung U, Nishishita T, Ebisu S, Usuda S, Mishiro S, Xanthoudakis S, Igarashi T, Ogata E. A redof factor protein, ref-1, is involved in negative gene regulation by extracellular calcium.  J Biol Chem. 1994;  269 27 855-27 862
    PubMedGoogle Scholar
  • 33 Kambe F, Nomura Y, Okamoto T, Seo H. Redox regulation of thyroid-transcription factors, Pax-8 and TTF-1, is involved in their increased DNA-binding activities by thyrotropin in rat thyroid FRTL-5 cells.  Mol Endocrinol. 1996;  10 801-812
    CrossrefPubMedGoogle Scholar
  • 34 Yanagita Y, Okajima F, Sho K, Nagamachi Y, Kondo Y. An adenosine derivative cooperates with TSH and Graves’ IgG to induce Ca2+ mobilization in single human thyroid cells.  Mol Cell Endocrinol. 1996;  118 47-56
    CrossrefPubMedGoogle Scholar
  • 35 Damante G, Tell G, di Lauro R. A unique combination of transcription factors controls differentiation of thyroid cells.  Progr Nucleic Acid Res Mol Biol. 2000;  66 307-356
    PubMedGoogle Scholar
  • 36 Xanthoudakis S, Curran T. Identification and characterization of Ref-1, a nuclear protein that facilitates AP-1 DNA-binding activity.  EMBO J. 1992;  11 653-665
    PubMedGoogle Scholar
  • 37 Shibasaki F, Price E R, Milan D, McKeon F. Role of kinases and the phosphatase calcineurin in the nuclear shuttling of transcription factor NF-AT4.  Nature. 1996;  382 370-373
    CrossrefPubMedGoogle Scholar

Prof. G. Damante

Dipartimento di Scienze e Tecnologie Biomediche, Università di Udine

Piazzale Kolbe 1 · 33100 Udine · Italy ·

Phone: + 39 (0432) 494374.

Fax: + 39 (0432) 494379.

Email: Gdamante@makek.dstb.uniud.it