TTF-2 stimulates expression of 17 genes, including one novel thyroid-specific gene which might be involved in thyroid development

• Published in: Molecular and cellular endocrinology Vol. 221; no. 1; pp. 33 - 46
• Main Authors: Hishinuma, Akira, Ohmika, Narumi, Namatame, Takashi, Ieiri, Tamio
• Format: Journal Article
• Language: English
• Published: Ireland Elsevier Ireland Ltd 30-06-2004
• Subjects: Amino Acid Sequence; Base Sequence; Cell Line; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Forkhead Transcription Factors; Gene Expression Profiling; Humans; Luciferase reporter assay; Membrane Proteins - genetics; Membrane Proteins - metabolism; Microarray analysis; Molecular Sequence Data; Oligonucleotide Array Sequence Analysis; Quantitative RT-PCR; Repressor Proteins - genetics; Repressor Proteins - metabolism; Subtractive cDNA library; Thyroid Gland - growth & development; Thyroid Gland - metabolism; Thyroid transcription factor-2; Tissue Distribution - genetics; Transcriptional Activation; Up-Regulation; Hypothetical protein FLJ23091; UDP- N-acetyl-alpha- d-galactosamine:polypeptide N-acetylgalactosaminyltransferase 1; Subtractive cDNA library; Quantitative RT-PCR; Tropomyosin 1; Collagen type 1 alpha-1; Integrin beta-1; Carboxypeptidase E; Retinal binding protein 7; Thyroid transcription factor-2; Luciferase reporter assay; Ribonuclease 6 precursor; Adenosine A1 receptor; Alpha synuclein; Thymosin beta-10; Annexin A5; P53-induced protein PIGPC1; Ring finger protein 29; Collagen type XI alpha-1; Latent transforming growth factor-beta-binding protein 1; Smooth muscle LIM protein; Microarray analysis
• ISSN: 0303-7207; 1872-8057
• DOI: 10.1016/j.mce.2004.04.003

Thyroid dysgenesis is the most frequent cause of congenital hypothyroidism, but its molecular pathophysiology is largely unknown. Our hypothesis that some genes downstream to thyroid transcription factor-2 (TTF-2) might be responsible for development of the thyroid prompted us to identify genes whose expression is stimulated by TTF-2. PCR products of cDNA clones obtained by a subtraction PCR method in TTF-2 expressing cell lines were screened with labeled cDNA by microarray analysis. We isolated 17 genes up-regulated by TTF-2, which were subsequently confirmed by quantitative reverse transcription-polymerase chain reaction (RT-PCR). One of them is a novel gene designated T1560 that showed a highly thyroid-specific expression pattern. Luciferase reporter assays showed that expression of all of the 14 genes tested was stimulated by both TTF-2 and TTF-1, another thyroid-specific transcription factor. Our results have important implications for understanding normal thyroid development as well as the molecular defects underlying thyroid dysgenesis.