GATA-4 gene organization and analysis of its promoter.

The mouse GATA-4 gene is separated by six introns, and this gene organization is conserved in rodents and man. The transcriptional start site of the GATA-4 gene is essentially the same in rat heart, stomach and testis, and in cultured cells expressing GATA-4 such as TM3, TM4, I-10 and P19.CL6 cells. The 5'-upstream of the GATA-4 gene is also conserved in rodents and man. We examined its promoter activity by means of luciferase reporter gene assay using testis-derived TM3 and TM4 cells. The GC-boxes and E-box located in the several tens of base pairs upstream of the transcriptional start sites of the GATA-4 gene were found to be critical for its promoter activity in these cells, consistent with the mode of transcription characteristics of the TATA-less promoter. P19.CL6 cells differentiate into beating cardiomyocytes upon induction by DMSO, accompanied by stimulation of the transcription of heart-specific genes including GATA-4. Interestingly, they exhibit increased luciferase reporter gene activity upon induction by DMSO. Both proximal tandem GC-boxes and the E-box are also contributed to the reporter gene activity in P19.CL6 cells.