Abstract
PCR-based methods and mobility shift competition assays were used to determine the basic biochemical features of the homeodomain transcription factor Distal-less 3 (Dlx3), including an optimal DNA binding site, the binding constant and dissociation rates of this protein. Expression of Dlx3 protein in either HeLa cells or Xenopus embryos resulted in strong activation of a model target gene construct containing three tandem copies of the Dlx3 binding site upstream from the TATA element. In addition, deletion analysis revealed that transcriptional activation by Dlx3 depends on two subdomains located on either side of the homeobox: removal of either subdomain resulted in complete loss of Dlx3 function. These observations provide new insight regarding the function of Dlx3 in vertebrate development and tissue differentiation and also suggest a mechanism for the dominant inheritance pattern of a hereditary disease resulting from mutation of the DLX3 gene in human.