Ubiquitin-dependent mechanisms of tissue-specific cell cycle control

该建议描述了一种识别组织特异性细胞模块的集成方法 循环控制尽管有明确的证据表明，在增殖和细胞周期中存在组织特异性， 组织特异性细胞周期调节对发育的重要性， 疾病的潜在机制尚未被发现。确定和 在增殖网络中的组织特异性模块的表征将不仅提供 深入了解多细胞生物的发展和肿瘤发生，但也奠定了 为发现组织特异性化疗药物奠定了基础。 由于泛素化在细胞周期控制中的关键作用， 泛素化酶的表达，泛素化的频繁失调， 癌症，和泛素化的酶的性质，我们将集中解剖组织- 对泛素化机制成员的特异性细胞周期调控。 在不同的细胞系中使用合成的致死性siRNA筛选，我们将鉴定 组织特异性细胞周期控制所需的泛素化酶。通过隔离 底物的这些酶独特的某一组织，我们将描绘途径 在这个组织中潜在的细胞周期调节。最后，通过开发和 可量化的泛素化分析，我们将分离小分子抑制剂， 进一步解剖增殖网络，并可作为新的 组织特异性化疗药物。 我们相信，这种综合方法利用基因发现，生物化学， 解剖和抑制剂鉴定的基础上化学生物学将提供深入的 深入了解细胞周期和发育调控，并为新的， 具有改进的组织特异性的创新化疗药物。

This proposal describes an integrated approach to identify modules of tissue-specific cell cycle control. Despite clear evidence for tissue-specificity in proliferation and the outstanding importance of tissue-specific cell cycle regulation for development and disease, the underlying mechanisms have not been uncovered. The identification and characterization of tissue-specific modules in proliferative networks will not only provide deep insight into development of multicellular organisms and tumorigenesis, but also lay the groundwork for the discovery of tissue-specific chemotherapeutics. Because of the crucial role of ubiquitination in cell cycle control, the tissue-specific expression of ubiquitination enzymes, the frequent misregulation of ubiquitination in cancer, and the enzymatic nature of ubiquitination, we will focus the dissection of tissue- specific cell cycle regulation on members of the ubiquitination machinery. Using synthetic lethality siRNA screens in different cell lines, we will identify ubiquitination enzymes required for tissue-specific cell cycle control. By isolating substrates of these enzymes unique to a certain tissue, we will delineate pathways underlying cell cycle regulation in this tissue. Finally, by developing and miniaturizing quantifiable ubiquitination assays, we will isolate small-molecule inhibitors that allow further dissection of proliferative networks and may serve as lead structures for novel tissue-specific chemotherapeutics. We are convinced that this integrated approach using genetic discovery, biochemical dissection, and inhibitor identification based on chemical biology will provide deep insights into cell cycle and developmental regulation and lay the groundwork for novel, innovative chemotherapeutics with improved tissue-specificity.