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Background  The molecular mechanisms leading to a fully differentiated thyrocite are still object of intense study even if it is well known that thyroglobulin, thyroperoxidase, NIS and TSHr are the marker genes of thyroid differentiation. It is also well known that Pax8, TTF-1, Foxe1 and Hhex are the thyroid-enriched transcription factors responsible for the expression of the above genes, thus are responsible for the differentiated thyroid phenotype. In particular, the role of Pax8 in the fully developed thyroid gland was studied in depth and it was established that it plays a key role in thyroid development and differentiation. However, to date the bases for the thyroid-enriched expression of this transcription factor have not been unraveled yet. Here, we report the identification and characterization of a functional thyroid-specific enhancer element located far upstream of the  Pax8  gene.    Results  We hypothesized that regulatory cis-acting elements are conserved among mammalian genes. Comparison of a genomic region extending for about 100 kb at the 5'-flanking region of the mouse and human  Pax8  gene revealed several conserved regions that were tested for enhancer activity in thyroid and non-thyroid cells. Using this approach we identified one putative thyroid-specific regulatory element located 84.6 kb upstream of the  Pax8  transcription start site. The  in silico  data were verified by promoter-reporter assays in thyroid and non-thyroid cells. Interestingly, the identified far upstream element manifested a very high transcriptional activity in the thyroid cell line PC Cl3, but showed no activity in HeLa cells. In addition, the data here reported indicate that the thyroid-enriched transcription factor TTF-1 is able to bind  in vitro  and  in vivo  the Pax8 far upstream element, and is capable to activate transcription from it.    Conclusions  Results of this study reveal the presence of a thyroid-specific regulatory element in the 5' upstream region of the  Pax8  gene. The identification of this regulatory element represents the first step in the investigation of upstream regulatory mechanisms that control  Pax8  transcription during thyroid differentiation and are relevant to further studies on  Pax8  as a candidate gene for thyroid dysgenesis.

Comparative genomics reveals a functional thyroid-specific element in the far upstream region of the PAX8 gene