Cell Cycle Dynamics that Ensure Genome Maintenance

确保基因组维持的细胞周期控制（库克） 总结 我们的研究计划旨在深入了解的基本架构和监管的 人类细胞分裂周期，特别关注DNA复制能力。完整和 整个哺乳动物基因组的有效复制需要数千个DNA 通过MCM解旋酶的DNA装载，复制起点被许可在G1期 配合物负载的MCM复合物使基因组位点能够进行复制起始 在S期。失去正常的原产地许可控制会导致基因组不稳定， 导致肿瘤发生、发育缺陷和退化。产地来源证的管制， 重要的是确保正常的基因组维护作为伴随机制， 复制叉的控制和稳定性或DNA修复，但原产地许可证的监管只是 部分理解。例如，如何在具有 G1期长度的显著不同，例如在发育过程中或癌基因激活后？ 在异源染色质环境中，原产地许可活动是如何分布的？怎么样 在进入和退出细胞周期的过渡期间控制原产地许可？这些悬而未决的 这些问题妨碍了对扩散控制的全面理解， 诊断和治疗以细胞增殖为标志的病理。 我们的长期目标是了解DNA复制起点许可是如何由 控制增殖和发育的细胞内和细胞外途径， 了解这些控制扰动的结果。我们目前和未来的项目是 组织成科学问题，分为两个中心目标：目标1）定义MCM如何 负载动力学调节G1进展，目标2）确定 建立和维持细胞周期退出到静止状态。近年来，我们不断创新， 使用定量活细胞和固定单细胞分析的实验工具和方法， 培养的人体细胞我们将这些工具与分子遗传学和生物化学联合收割机相结合。我们 专注于揭示分子机制及其相互关系，然后测试 扰乱这些机制的后果。我们之前的努力产生了一个一致的流 基础科学发现和进步的领域和科学劳动力。的 成功实现我们的中心目标的影响是确定以前未探索的机制 在哺乳动物细胞周期和探测基因组所需的分子事件的动力学 上维护

CELL CYCLE CONTROLS THAT ENSURE GENOME MAINTENANCE (COOK) SUMMARY Our research program seeks insight into the fundamental architecture and regulation of the human cell division cycle, with specific focus on DNA replication competence. Complete and efficient duplication of an entire mammalian genome requires that many thousands of DNA replication origins become licensed in G1 phase through the DNA loading of MCM helicase complexes. Loaded MCM complexes render genomic loci competent for replication initiation during S phase. Loss of normal origin licensing control causes genome instability, which can cause oncogenesis, developmental defects, and degeneration. Origin licensing control is as important for ensuring normal genome maintenance as companion mechanisms such as replication fork control and stability or DNA repair, but the regulation of origin licensing is only partly understood. For example, how is complete origin licensing achieved in cells with dramatically different G1 lengths, such as during development or after oncogene activation? How is origin licensing activity distributed in a heterogenous chromatin environment? How is origin licensing controlled during transitions into and out of the cell cycle? These unanswered questions preclude the comprehensive understanding of proliferation control needed to diagnose and treat pathologies in which cell proliferation is a hallmark. Our long-term goal is to understand how DNA replication origin licensing is governed by intracellular and extracellular pathways that control proliferation and development and to understand the outcomes of perturbations to those controls. Our current and future projects are organized into scientific questions clustered into two central goals: Goal 1) Define how MCM loading dynamics regulate G1 progression, Goal 2) Determine the molecular events that establish and maintain cell cycle exit to quiescence. In recent years, we developed innovative experimental tools and approaches using quantitative live cell and fixed single cell analyses in cultured human cells. We combine these tools with molecular genetics and biochemistry. We focus on uncovering molecular mechanisms and their inter-relationships, and then test the consequences of perturbing those mechanisms. Our prior efforts produced a consistent stream of basic scientific discoveries and advances for both the field and the scientific workforce. The impacts of success towards our central goals are to define previously unexplored mechanisms in the mammalian cell cycle and to probe the dynamics of molecular events required for genome maintenance.