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

G. Tell; A. Pines; M. Pandolfi; A. V. D’Elia; D. Donnini; R. Lonigro; G. Manzini; D. Russo; C. Di Loreto; G. Damante

doi:10.1055/s-2002-33258

Hormone and Metabolic Research, Table of Contents

Horm Metab Res 2002; 34(6): 303-310
DOI: 10.1055/s-2002-33258

Original Basic

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

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

Full Text

References

1 Nakamura H, Nakamura K, Yodoi J. Redox regulation of cellular activation. Annu Rev Immunol. 1997; 15 351-369
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
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
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
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
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
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
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
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
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
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
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
13 Ambesi-Imbiombato F S, Coon H G. Thyroid cells in culture. Int Rev Cytol. 1979; 10 163-172
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
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
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
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
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
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
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
21 Nunez J, Pommier J. Formation of thyroid hormones. Vitam Horm. 1982; 39 175-229
22 Bjorkman U, Ekholm R. Accelerated exocytosis and H₂O₂ generation in isolated thyroid follicles enhance protein iodination. Endocrinology. 1988; 130 393-399
23 Sies H. Glutathione and its role in cellular functions. Free Radic Biol Med. 1999; 27 916-921
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
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
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
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
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
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
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
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
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
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
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
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
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
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

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