Transcriptional activation by the homeodomain protein Distal-less 3
Transcriptional activation by the homeodomain protein Distal-less 3
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DOI：
10.1093/nar/27.3.764
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发表时间：
1999-02-01
影响因子：
14.9
通讯作者：
Sargent, TD
中科院分区：
文献类型：
--
作者：
Feledy, JA;Morasso, MI;Sargent, TD
基于 PCR 的方法和迁移率变化竞争测定用于确定同源域转录因子 Distal-less 3 (Dlx3) 的基本生化特征，包括最佳 DNA 结合位点、该蛋白质的结合常数和解离速率。 Dlx3 蛋白在 HeLa 细胞或非洲爪蟾胚胎中的表达导致模型靶基因构建体的强烈激活，该模型靶基因构建体包含 TATA 元件上游 Dlx3 结合位点的三个串联拷贝。此外，删除分析表明，Dlx3 的转录激活取决于位于同源盒两侧的两个子结构域：删除任一子结构域都会导致 Dlx3 功能完全丧失。这些观察结果为Dlx3在脊椎动物发育和组织分化中的功能提供了新的见解，并且还提出了人类DLX3基因突变引起的遗传性疾病的显性遗传模式的机制。
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.