Polymerase theta, genome instability, and cancer

项目概要/描述 聚合酶θ（Pol q，基因名称Polq）在许多遗传性乳腺癌中是必需的，但Pol q的缺失是遗传性乳腺癌的重要组成部分。 在大多数正常细胞中耐受良好。因此，在发展中出现了许多令人兴奋的情况， 用于癌症治疗的Pol q的靶向抑制剂。然而，我们对它的生物学作用知之甚少， 这一大型多领域因素的作用机制。因此，不可能调和不同的， Pol q缺失对突变、染色体畸变和细胞存活的明显背景依赖性影响， 并且安全、有效地靶向Pol q用于治疗在很大程度上受到了挫折。 在总体目标1中，全长Pol q和Pol q结构域有助于修复的机制将是 其特征在于使用生物化学、结构、遗传和生物物理成像的平行分析 我们的方案可用的方法。完整的路径重建和可视化是一个目标。总体目标2 我们将研究通常参与Pol q的细胞环境。在总体目标3中，我们将整合见解 从这些其他目的中获得，以开发在癌症治疗中更安全、有效地靶向Pol q的基本原理。 研究工作将在三个核心框架内的计划项目内高度协调 设施基质、蛋白质和实验将与所有项目一起设计，并通过 通过管理核心A进行反馈。核心B和细胞系构建将支持蛋白纯化 核心C 科学项目负责人具有互补的专业知识：Dale Ramsden博士（分子生物学;项目 1），Gaorav Gupta（癌细胞生物学;项目1），Richard Wood（生物化学，项目2）Sylvie Doublié （结构生物学和机制;项目3）和Eli Rothenberg（生物物理学;项目4）。这套 互补的专业知识促进了对整个团队有益的想法的交叉，并使工作 只有通过计划项目赠款才能实现。这支球队也已经有了很长的路要走 富有成效的合作记录，并在此计划项目将有效和协同工作， 实现计划的目标。该计划项目所取得的成果将提供一个基本的 在理解TMEJ的分子机制方面取得了进展，并将为 通过抑制Pol θ设计新的癌症疗法。

PROJECT SUMMARY/DESCRIPTION Polymerase theta (Pol q, gene name Polq) is essential in many hereditary breast cancers, yet loss of Pol q is well tolerated in most normal cells. As a consequence, there has been much excitement in the development of targeted inhibitors of Pol q for cancer therapy. However, we know little about the biological role and mechanism of action for this large, multi-domain factor. It has thus not been possible to reconcile the disparate, apparently context-dependent impacts of Pol q loss on mutation, chromosome aberration, and cell survival, and the safe, effective targeting of Pol q for therapy has been largely frustrated. In overall Aim 1, the mechanism by which full-length Pol q and Pol q domains contribute to repair will be characterized by parallel analysis using the biochemical, structural, genetic and biophysical imaging approaches available to our program. Full pathway reconstitution and visualization is a goal. In Overall Aim 2 we will investigate the cellular contexts that normally engage Pol q. In Overall Aim 3 we will integrate insights gained from these other Aims to develop rationales for safer, effective targeting of Pol q in cancer therapy. The research work will be highly coordinated within the Program Project in a framework with three Core facilities. Substrates, proteins, and experiments will be designed with all Projects and constantly monitored with feedback via Administrative Core A. Protein purification will be supported by Core B, and cell line construction by Core C. The scientific project leaders have complementary expertise: Drs. Dale Ramsden (molecular biology; Project 1), Gaorav Gupta (cancer cell biology; Project 1), Richard Wood (biochemistry, Project 2) Sylvie Doublié (structural biology and mechanism; Project 3), and Eli Rothenberg (biophysics; Project 4). This ensemble of complementary expertise fosters cross-fertilization of ideas beneficial to the whole team, and makes work possible that can only be accomplished by a Program Project grant. This team also already has a long track record of productive collaboration, and within this program project will work effectively and synergistically to accomplish the Program’s goals. The results obtained by this Program Project will provide a fundamental advance in the understanding of the molecular mechanisms underpinning TMEJ, and will pave the way for the design of novel cancer therapy via Pol θ inhibition.