Interpreting and Deploying Genomic Information During Animal Development

总结 该项目的长期目标是了解大转录因子的成员如何 家族，共享 DNA 结合域并且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。该提案的主要重点是 同源域转录因子 Hox 家族，在动物发育过程中发挥着关键作用 脊椎动物和动物的胚胎发生、器官发育和附肢规范 无脊椎动物。当这些转录因子错误调节其靶基因时，要么是由于 其蛋白质编码序列或其靶 DNA 结合位点发生突变，发育 可能会导致缺陷和许多人类疾病，从癌症到糖尿病。为下一步的资金筹措 在此期间，该项目将弥合理解这些因素具体如何变化之间的差距 在体外识别它们的结合位点（通常与辅因子结合），以及它们如何执行 它们在体内的特定功能。这将通过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质及其在活体中的结合位点 细胞。额外的努力将是识别和表征新发现的新型辅助因子和 可以改变 DNA 的机制，例如 Hox 蛋白的翻译后修饰 体内结合特异性和/或转录调节活性。在所有情况下，这些 实验将基于识别和理解 Hox 蛋白如何调节其 真正的目标基因，使他们能够协调整个发育计划，例如 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.