Redox Effector Factor-1 Regulates the Activity of Thyroid Transcription Factor 1 by Controlling the Redox State of the N Transcriptional Activation Domain*

Thyroid transcription factor 1 (TTF-1) is a homeodomain-containing transcriptional regulator responsible for the activation of thyroid- and lung-specific genes. It has been demonstrated that its DNA binding activity is redox-regulated in vitro through the formation of dimers and oligomeric species. In this paper, we demonstrate that the redox regulation mainly involves a Cys residue (Cys87), which resides out of the DNA binding domain, belonging to the N-transactivation domain. In fact, the oxidized form of a truncated TTF-1 (containing the N-transactivation domain and the DNA-binding domain, here called TTF-1N-HD) looses specific DNA binding activity. Since most of the oxidized TTF-1N-HD is in a monomeric form, these data indicate that the redox state of Cys87 may control the DNA-binding function of the homeodomain, suggesting that Cys87 could play an important role in determining the correct folding of the homeodomain. By using gel retardation and transient transfection assays, we demonstrate that the redox effector factor-1 (Ref-1) mediates the redox effects on TTF-1N-HD binding and that it is able to modulate the TTF-1 transcriptional activity. Glutathione S-transferase pull-down experiments demonstrate the occurrence of interaction between Ref-1 and TTF-1N-HD. Having previously demonstrated that Ref-1 is able to modulate the transcriptional activity of another thyroid-specific transcription factor (Pax-8), our data suggest that Ref-1 plays a central role in the regulation of thyroid cells.

[1]  H. E. Marshall,et al.  Nitrosation and oxidation in the regulation of gene expression , 2000, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[2]  G. Tell,et al.  TSH controls Ref-1 nuclear translocation in thyroid cells. , 2000, Journal of molecular endocrinology.

[3]  R. Barouki,et al.  The repression of nuclear factor I/CCAAT transcription factor (NFI/CTF) transactivating domain by oxidative stress is mediated by a critical cysteine (Cys-427). , 2000, The Biochemical journal.

[4]  G. Tell,et al.  An 'environment to nucleus' signaling system operates in B lymphocytes: redox status modulates BSAP/Pax-5 activation through Ref-1 nuclear translocation. , 2000, Nucleic acids research.

[5]  C. Prives,et al.  Ref‐1 regulates the transactivation and pro‐apoptotic functions of p53 in vivo , 1999, The EMBO journal.

[6]  P. Chanson,et al.  Biochemical characterization of a Ca2+/NAD(P)H-dependent H2O2 generator in human thyroid tissue. , 1999, Biochimie.

[7]  G. Tell,et al.  Ref-1 controls pax-8 DNA-binding activity. , 1998, Biochemical and biophysical research communications.

[8]  A. Scaloni,et al.  Redox Potential Controls the Structure and DNA Binding Activity of the Paired Domain* , 1998, The Journal of Biological Chemistry.

[9]  C. Ramana,et al.  Activation of apurinic/apyrimidinic endonuclease in human cells by reactive oxygen species and its correlation with their adaptive response to genotoxicity of free radicals. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[10]  G. Tell,et al.  Structural and functional properties of the N transcriptional activation domain of thyroid transcription factor-1: similarities with the acidic activation domains. , 1998, The Biochemical journal.

[11]  R. Hammer,et al.  Genetic interaction between HAP1/REF-1 and p53 , 1997, Nature Genetics.

[12]  H. Seo,et al.  Increase in Ref-1 mRNA and protein by thyrotropin in rat thyroid FRTL-5 cells. , 1997, Biochemical and biophysical research communications.

[13]  T. Curran,et al.  Identification of redox/repair protein Ref-1 as a potent activator of p53. , 1997, Genes & development.

[14]  M. Zannini,et al.  Redundant Domains Contribute to the Transcriptional Activity of the Thyroid Transcription Factor 1 (*) , 1995, The Journal of Biological Chemistry.

[15]  M. Kelley,et al.  Expression in Escherichia coli of a rat cDNA encoding an apurinic/apyrimidinic endonuclease. , 1995, Mutation research.

[16]  D. Bradley,et al.  Quantification of DNA-protein interaction by UV crosslinking. , 1995, Nucleic acids research.

[17]  M. Arnone,et al.  The DNA Binding Activity and the Dimerization Ability of the Thyroid Transcription Factor I Are Redox Regulated (*) , 1995, The Journal of Biological Chemistry.

[18]  J. Whitsett,et al.  Lung Cell-specific Expression of the Murine Surfactant Protein A (SP-A) Gene Is Mediated by Interactions between the SP-A Promoter and Thyroid Transcription Factor-1 (*) , 1995, The Journal of Biological Chemistry.

[19]  S. Formisano,et al.  Sequence-specific DNA recognition by the thyroid transcription factor-1 homeodomain. , 1994, Nucleic acids research.

[20]  C. Robson,et al.  Identification of residues in the human DNA repair enzyme HAP1 (Ref-1) that are essential for redox regulation of Jun DNA binding , 1993, Molecular and cellular biology.

[21]  M. Zannini,et al.  Multiple mechanisms of interference between transformation and differentiation in thyroid cells , 1992, Molecular and cellular biology.

[22]  T. Curran,et al.  Redox activation of Fos‐Jun DNA binding activity is mediated by a DNA repair enzyme. , 1992, The EMBO journal.

[23]  R. di Lauro,et al.  Cell-type-specific expression of the rat thyroperoxidase promoter indicates common mechanisms for thyroid-specific gene expression , 1992, Molecular and cellular biology.

[24]  E. Laurent,et al.  The H2O2-generating system modulates protein iodination and the activity of the pentose phosphate pathway in dog thyroid. , 1991, Endocrinology.

[25]  M. Mattéi,et al.  Thyroid nuclear factor 1 (TTF‐1) contains a homeodomain and displays a novel DNA binding specificity. , 1990, The EMBO journal.

[26]  R. Lauro,et al.  The tissue-specific expression of the thyroglobulin gene requires interaction between thyroid-specific and ubiquitous factors. , 1990, European journal of biochemistry.

[27]  T. Curran,et al.  Redox regulation of fos and jun DNA-binding activity in vitro. , 1990, Science.

[28]  M. Yoshida,et al.  Multiple cDNA clones encoding nuclear proteins that bind to the tax‐dependent enhancer of HTLV‐1: all contain a leucine zipper structure and basic amino acid domain. , 1990, The EMBO journal.

[29]  P. V. von Hippel,et al.  Calculation of protein extinction coefficients from amino acid sequence data. , 1989, Analytical biochemistry.

[30]  R. Lauro,et al.  A thyroid‐specific nuclear protein essential for tissue‐specific expression of the thyroglobulin promoter. , 1989, The EMBO journal.

[31]  D. George,et al.  Structural and functional characterization of the promoter region of the mouse c-Ki-ras gene , 1987, Molecular and cellular biology.

[32]  I. Verma,et al.  Modulation of c-"os gene transcription by negative and positive cellular factors , 1987, Nature.

[33]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[34]  A. van der Eb,et al.  A new technique for the assay of infectivity of human adenovirus 5 DNA. , 1973, Virology.

[35]  G. Tell,et al.  A unique combination of transcription factors controls differentiation of thyroid cells. , 2001, Progress in nucleic acid research and molecular biology.