CELL CYCLE CONTROLS THAT ENSURE GENOME MAINTENANCE

确保基因组维持的细胞周期控制(库克) 摘要 我们的研究计划旨在深入了解 人类细胞分裂周期，特别关注DNA复制能力。完成并 整个哺乳动物基因组的有效复制需要成千上万的DNA 通过MCM解旋酶的DNA负载，复制起始点在G1期获得许可 复合体。负载的MCM复合体使基因组位点具有复制起始能力 在S阶段。失去正常的原产地许可控制会导致基因组不稳定，这可能 导致肿瘤发生、发育缺陷和退化。产地来源许可证管制为 对于确保正常的基因组维护非常重要，因为伴随机制包括 复制分叉的控制和稳定或DNA修复，但对原产地许可的监管只是 部分地理解了。例如，如何在细胞中实现完整的原产地许可 明显不同的G1长度，例如在发育过程中或在癌基因激活之后？ 在异质染色质环境中，原产地许可活动是如何分布的？你好吗？ 在进入和退出细胞周期的过程中，原产地许可受到控制吗？这些未回答的问题 这些问题妨碍了对扩散控制的全面了解，这是 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复制起源许可是如何由 控制增殖和发育的细胞内和细胞外途径以及 了解对这些控制进行扰动的结果。我们目前和未来的项目是 组织成两个中心目标的科学问题：目标1)如何定义MCM 负荷动态调节G1进程，目标2)决定分子事件 建立和维持细胞周期退出到静止状态。近年来，我们开发了创新的 使用定量活细胞和固定单细胞分析的实验工具和方法 培养的人类细胞。我们将这些工具与分子遗传学和生物化学结合起来。我们 重点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致的结果 为该领域和科学工作者提供基本的科学发现和进步。这个 成功对我们中心目标的影响是定义以前未曾探索过的机制 在哺乳动物细胞周期中并探索基因组所需分子事件的动力学 维修。

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.