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Papers In Press, published online ahead of print April 10, 2002
J. Biol. Chem, 10.1074/jbc.M201737200
Submitted on February 20, 2002
Revised on April 10, 2002
Accepted on April 10, 2002
Department of Biological Sciences, The University of Tulsa, Tulsa, OK 74104
Corresponding Author: brad-amendt{at}utulsa.edu
Three major PITX2 isoforms are differentially expressed in humans, mice, zebrafish, chick and frog tissues. To demonstrate differential regulation of gene expression by these isoforms we used three different promoters and three cell lines. Transient transfection of CHO, HeLa and LS-8 cell lines reveal differences in PITX2A and PITX2C activation of the PLOD1 and Dlx2 promoters, however, PITX2B is inactive. In contrast, PITX2B actives the pituitary specific Prolactin promoter at higher levels than either PITX2A or PITX2C. Interestingly, co-transfection of either PITX2A or PITX2C with PITX2B results in a synergistic activation of the PLOD1 and Dlx2 promoters. Furthermore, PITX2 isoforms have different transcriptional activity dependent upon the cells used for transfection analysis. We have isolated a fourth PITX2 isoform (PITX2D) expressed only in humans, which acts to suppress the transcriptional activity of the other PITX2 isoforms. EMSA and GST pull-down experiments demonstrate that all isoforms interact with PITX2D, and PITX2B forms heterodimeric complexes with PITX2A and PITX2C. Our research provides a molecular basis for differential gene regulation through the expression of PITX2 isoforms. PITX2 isoform activities are both promoter and cell specific and our data reveal new mechanisms for PITX2 regulated gene expression.
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