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.

遗传信息的遗传要求将DNA复制并包装成分子“适配器”或“手柄”的形式，从而使DNA有效地分离为子细胞。 但是，这两个继承步骤之间的关系知之甚少。 该项目将奠定有关DNA复制协调的科学原则的基础，以及在细胞分裂期间其随后的隔离的这些早期步骤。 这项工作还将有助于对这些DNA代谢过程如何实时发生在活细胞中的新理解。 该项目将导致以色列和美国的研究生进行跨学科的教育，以批判性地评估细胞生物学，数学，统计和热力学方面的实验技术。 为了进一步扩大该项目的影响，研究小组的主要研究者和成员将为几项外展活动做出贡献，以将DNA继承的复杂性和优雅传递给公众。 Pi Bloom将在Kenan Fellows计划中指导教师领导计划，这是一项北卡罗来纳州的STEM教师领导力计划。 为了促进代表性不足的科学群体的更广泛参与，PI将指导科学和数学成就和足智运动轨道（SMART）计划，由NC-LOUIS Stokes for Millinal参与（NC-LSAMP）在科学领域赞助。 在真核细胞分裂期间，对气体组件和染色体隔离至关重要。在每个细胞周期中，必须拆卸中心粒DNA和动力学蛋白的复合物，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，但对于通过丝粒区域的补充进展与在这些位点的功能动物学组装之间的相互作用知之甚少。 该项目将结合遗传学，细胞生物学，活细胞显微镜和计算方法，以研究DNA复制，Centromere建立，动力学组装和染色体分离之间的关系之间的关系。 具体而言，将使用通过主动和不活跃的中心层面区域来研究DNA构型对Centromere组装的重要性，计算机模拟和最近开发的活细胞成像方法。 这种集成方法应用于多种突变菌株和条件，将使该项目能够对Centromeric DNA复制，基因组稳定性，染色体分离和细胞分裂的不同方面进行新的启示。 这将提供有关中心区域的转录如何影响动力学大会的批判性见解。该协作美国/以色列的项目得到了美国国家科学基金会和以色列双性科学基金会的支持。该奖项反映了NSF的法定使命，并认为通过基金会的知识优点和广泛的crietia crietia criperia criperia criperia criperia criperia criperia criperia cribitia criperia criperia criperia criperia recteria均值得通过评估。