Interpreting and Deploying Genomic Information During Animal Development

摘要 这个项目的长期目标是了解大转录因子的成员是如何 共享DNA结合域的家族，通常具有非常相似的DNA结合 特异性，在体内执行高度特异性的功能。这项提案的主要重点是 HOX同源结构域转录因子家族，在动物生长发育过程中发挥重要作用 脊椎动物的胚胎发生、器官发育和附肢规格 无脊椎动物。当这些转录因子错误调节它们的目标基因时，要么是由于 其蛋白质编码序列或其目标DNA结合位点的突变，发育 缺陷和许多人类疾病，从癌症到糖尿病，都可能导致。用于下一笔资金 在此期间，该项目将弥合了解这些因素如何具体 在体外识别它们的结合部位，通常与辅因子结合，以及它们是如何执行的 它们在体内的特定功能。这将使用全基因组方法来完成， 标记和突变的HOX因子，并通过可视化这些蛋白及其在生物中的结合位置 细胞。更多的努力将是确定和表征新发现的新辅助因子和 机制，例如HOX蛋白的翻译后修饰，可以改变它们的DNA 体内结合特异性和/或转录调节活性。在所有情况下，这些 实验的基础将是识别和理解Hox蛋白是如何调节它们的 真正的靶向基因，使它们能够协调整个发育程序，如 随着腿和翅膀的发育。

Summary The long term goal of this project is to understand how members of large transcription factor families, which share a DNA binding domain and typically have very similar DNA binding specificities, execute highly specific functions in vivo. The primary focus of this proposal is on the Hox family of homeodomain transcription factors, which play critical roles during animal embryogenesis, organ development, and appendage specification in both vertebrates and invertebrates. When these transcription factors mis-regulate their target genes, either due to mutations in their protein-coding sequences or in their target DNA binding sites, developmental defects and many human diseases, from cancer to diabetes, can result. For the next funding period this project will bridge the gap between understanding how these factors specifically recognize their binding sites in vitro, often in conjunction with cofactors, and how they execute their specific functions in vivo. This will be accomplished using whole genome approaches with tagged and mutant Hox factors and by visualizing these proteins and their binding sites in living cells. Additional efforts will be to identify and characterize newly discovered novel cofactors and mechanisms, such as post-translational modifications of Hox proteins, that can alter their DNA binding specificities and/or transcriptional regulatory activities in vivo. In all cases, these experiments will be grounded in identifying and understanding how Hox proteins regulate their bona fide target genes, which allow them to orchestrate entire developmental programs, such as the development of legs and wings.