Odd-skipped related 1 gene expression is regulated by Runx2 and Ikzf1 transcription factors

Odd-skipped related 1 (Osr1) gene encodes a zinc-finger transcription factor that plays important roles in embryonic, heart, and urogenital development, however, it is unknown how its expression is regulated. In this study, we analyzed the promoter region of Osr1 to elucidate its regulation mechanism. The mouse Osr1 promoter region was cloned and characterized, and found to have two repressor elements in the -4504/-2766 and -1616/-109 regions, and two enhancer elements in the -2766/-1616 and -109/+199 regions. Several Runx2 and Ikzf1 binding sites were found in both mouse and human Osr1 promoters. Osr1 promoter activity was suppressed by cotransfection with Runx2- and Ikzf1-expressing vectors in a dose-dependent manner. Electrophoresis mobility shift assays showed that purified Runx2 bound to proximal (-611/-606) Runx2 binding motifs and that Ikzf1 bound to proximal (-1652/-1644) Ikzf1 binding motifs. Chrosmatin immunoprecipitation demonstrated that Runx2 bound to both the distal (-3047/-3042) and proximal regions, and that Ikzf1 bound to both the far-distal (-3036/-3028) and proximal elements. These findings indicate that Osr1 expression is regulated by Runx2 and Ikzf1, which are known as master-gene of osteogenesis and hematopoiesis, respectively.