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J. Biol. Chem., Vol. 278, Issue 5, 3395-3402, January 31, 2003
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From the Pax genes encode for transcription factors
essential for tissue development in many species. Pax8, the only
member of the family expressed in the thyroid tissue, is involved in
the morphogenesis of the gland and in the transcriptional regulation of
thyroid-specific genes. TTF-1, a homeodomain-containing factor, is also
expressed in the thyroid tissue and has been demonstrated to play a
role in thyroid-specific gene expression. Despite the presence of Pax8 and TTF-1 also in a few other tissues, the simultaneous expression of
the two transcription factors occurs only in the thyroid, supporting the idea that Pax8 and TTF-1 might cooperate to influence
thyroid-specific gene expression. In this report, we describe a
physical and functional interaction between these two factors. The
fusion protein GST-Pax8 is able to bind TTF-1 present in thyroid or in
non-thyroid cell extracts, and by using bacterial purified TTF-1 we
demonstrate that the interaction is direct. By co-immunoprecipitation,
we also show that the interaction between the two proteins occurs in vivo in thyroid cells. Moreover, Pax8 and TTF-1 when
co-expressed in HeLa cells synergistically activate Tg gene
transcription. The synergism requires the N-terminal activation domain
of TTF-1, and deletions of Pax8 indicate that the C-terminal domain of
the protein is involved. Our results demonstrate a
functional cooperation and a physical interaction between
transcription factors of the homeodomain-containing and of the
paired domain-containing gene families in the regulation of
tissue-specific gene expression.
The Paired Domain-containing Factor Pax8 and the
Homeodomain-containing Factor TTF-1 Directly Interact and
Synergistically Activate Transcription*
,,,,¶
Istituto di Endocrinologia ed Oncologia
Sperimentale, CNR and Department Biologia e Patologia Cellulare e
Molecolare, Via Pansini 5, 80131 Napoli and § Laboratory of
Biochemistry and Molecular Biology, Stazione Zoologica A. Dohrn, Villa
Comunale, 80121 Napoli, Italy
*
This work was supported by grants from the Consiglio
Nazionale delle Ricerche "Target Project on Biotechnology" and the
Ministero per l'Universitá e la Ricerca Scientifica (MIUR-URST)
Project "Molecular Mechanisms Responsible for Thyroid Cells
Differentiation."The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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