NSF-BSF: Defining the relationship between DNA replication kinetics and macromolecular protein assembly at the centromere

NSF-BSF：定义 DNA 复制动力学与着丝粒大分子蛋白质组装之间的关系

• 批准号: 1929114
• 负责人: Kerry Bloom
• 金额: $ 90万
• 依托单位: University of North Carolina at Chapel Hill
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1929114&HistoricalAwards=false
• 关键词: NSF; BSF; Defining; relationship; between

The inheritance of genetic information requires that DNA be replicated and packaged into a form with molecular "adaptors" or "handles" that allow the DNA to be efficiently segregated into daughter cells. However, little is known about the relationship between these two steps of inheritance. This project will lay the foundation of scientific principles governing the coordination of DNA replication and these early steps of its subsequent segregation during cell division. This work will also contribute new understandings of how these DNA metabolic processes occur in real-time within a living cell. The project will result in the interdisciplinary education of graduate students in Israel and the US, cross-training them to critically evaluate experimental techniques in cell biology, mathematics, statistics, and thermodynamics. To further broaden the impact of the project, the Principal Investigator and members of the research groups will contribute to several outreach events to convey the complexity and elegance of DNA inheritance to the general public. PI Bloom will mentor in the Kenan Fellows Program for Teacher leadership, a well-established STEM teacher leadership program in North Carolina. To facilitate broader participation of underrepresented groups in science, the PI will mentor in the Science and Math Achievement and Resourcefulness Track (SMART) program sponsored by the NC-Louis Stokes Alliance for Minority Participation (NC-LSAMP) in science. Centromeres are essential for kinetochore assembly and chromosome segregation during the eukaryotic cell division. At each cell cycle, the complex of centromere DNA and kinetochore proteins must be disassembled to allow DNA replication and then reassembled to allow microtubule attachment and chromosome segregation. Despite extensive studies, little is known regarding the interplay between replisome progression through centromeric regions and the assembly of functional kinetochores at these sites. This project will combine genetics, cell biology, live-cell microscopy, and computational methods to study the relationship between DNA replication, centromere establishment, kinetochore assembly, and chromosome segregation during mitosis. Specifically, to investigate the importance of DNA configurations to centromere assembly, computer simulations and recently developed live-cell imaging approaches for monitoring replication fork progression through active and inactive centromeric regions will be used. This integrative approach, applied to a variety of mutant strains and conditions, will enable the project to shed new light on different aspects of centromeric DNA replication, genome stability, chromosome segregation and cell division. This will provide critical insight into how transcription of centromeric regions affects kinetochore assembly.This collaborative US/Israel project is supported by the US National Science Foundation and the Israeli Binational Science Foundation.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

遗传信息的遗传需要DNA被复制并包装成具有分子“接头”或“手柄”的形式，使DNA能够有效地分离到子细胞中。 然而，人们对这两个遗传步骤之间的关系知之甚少。 该项目将奠定基础的科学原则，管理协调的DNA复制和这些早期的步骤，其随后的分离在细胞分裂。 这项工作还将有助于对这些DNA代谢过程如何在活细胞内实时发生的新理解。 该项目将导致以色列和美国研究生的跨学科教育，交叉培训他们批判性地评估细胞生物学，数学，统计学和热力学的实验技术。 为了进一步扩大该项目的影响，首席研究员和研究小组的成员将参与几项外展活动，向公众传达DNA遗传的复杂性和优雅性。 PI布鲁姆将在凯南研究员教师领导计划，一个完善的干在北卡罗来纳州教师领导计划导师。 为了促进代表性不足的群体更广泛地参与科学，PI将指导由NC-Louis Stokes少数民族参与联盟（NC-LSAMP）赞助的科学和数学成就和资源轨道（SMART）计划。 着丝粒是真核细胞分裂过程中着丝粒组装和染色体分离的关键。在每个细胞周期中，着丝粒DNA和动粒蛋白的复合物必须分解以允许DNA复制，然后重新组装以允许微管附着和染色体分离。尽管进行了广泛的研究，很少有人知道复制体之间的相互作用，通过着丝粒区域和组装的功能kinetochores在这些网站。 本计画将结合联合收割机遗传学、细胞生物学、活细胞显微镜及计算方法，研究有丝分裂过程中DNA复制、着丝粒建立、动粒组装与染色体分离之间的关系。 具体而言，调查DNA构型的重要性，着丝粒组装，计算机模拟和最近开发的活细胞成像方法，通过活跃和不活跃的着丝粒区域监测复制叉进展将被使用。 这种综合方法适用于各种突变株和条件，将使该项目能够对着丝粒DNA复制，基因组稳定性，染色体分离和细胞分裂的不同方面提供新的见解。 这将为着丝粒区域的转录如何影响动粒组装提供重要的见解。这个美国/以色列合作项目得到了美国国家科学基金会和以色列两国科学基金会的支持。这个奖项反映了NSF的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。