Understanding and exploiting DNA topoisomerases in cancer biology

摘要 DNA拓扑结构的适当控制对DNA的稳定性和流动性有重大影响。 遗传信息。本申请集中于II型DNA拓扑异构酶， 调节DNA超螺旋和去除染色体的分子机器 通过催化一个DNA双链体的ATP依赖性运输， 另II型拓扑异构酶在癌症生物学中发挥着重要作用， 都能维持和破坏基因组的完整性;它们也被证明是 治疗癌症 我们过去对真核拓扑异构酶II（topo II）的研究开辟了新的研究领域 了解癌症病因和改善癌症治疗的途径。本 该应用程序将为该领域的关键问题提供突破性的解决方案，包括如何 某些类型的抗拓扑异构酶II药物作用于酶，拓扑异构酶II是如何定位到关键部位的 它解决潜在有害的染色体拓扑结构的行动，以及如何 异常的Topo II活性可促进DNA损伤和遗传不稳定性。我们还将 调查创新的概念和高度重要的调查线提出了我们的新 研究发现，如TCA循环产生的代谢物如何控制拓扑结构II的功能。 我们的方法的特点是全面融合了生物化学，结构， 计算的、基于细胞的和化学生物学方法。影响力大的成果 将包括定义拓扑蛋白II如何适当地与染色质和伴侣蛋白定位 以减轻其天然的DNA损伤潜力，建立如何特异性的抗拓扑异构酶， II剂可以被改进以增强它们在癌症治疗中的效用，并且揭示了 II型拓扑异构酶中天然氨基酸序列变异使其不稳定的可能性 人类染色体和作为癌症驱动程序。过去的进展和未发表的研究结果 确定我们计划目标的可行性。

ABSTRACT The appropriate control of DNA topology has a major impact on the stability and flow of genetic information. The present application focuses on type II DNA topoisomerases, molecular machines that modulate DNA supercoiling and remove chromosome entanglements by catalyzing the ATP-dependent transport of one DNA duplex through another. Type II topoisomerases play a frontline role in cancer biology as factors that can both maintain and disrupt genome integrity; they are also demonstrated drug targets for treating cancer. Our past research on eukaryotic topoisomerase II (topo II) has opened up new research avenues for understanding cancer etiology and improving cancer treatment. The present application will deliver groundbreaking solutions to key problems in the field, including how certain classes of anti-topo II drugs act on the enzyme, how topo II is localized to key sites of action where it resolves potentially deleterious chromosomal topologies, and how aberrant topo II activity can promote DNA damage and genetic instability. We will also investigate innovative concepts and highly significant lines of inquiry raised by our new findings, such as how metabolites produced by the TCA cycle control topo II function. Our approach is distinguished by a comprehensive blend of biochemical, structural, computational, cell-based, and chemical biology methodologies. High-impact outcomes will include defining how topo II appropriately localizes with chromatin and partner proteins to mitigate its natural DNA-damaging potential, establishing how the specificity of anti-topo II agents can be improved to enhance their utility in cancer treatment, and revealing the potential for natural amino-acid sequence variation in type II topoisomerases to destabilize human chromosomes and act as cancer drivers. Past progress and unpublished findings establish the feasibility of our planned goals.