Molecular Analysis of Chromosome Segregation

染色体分离的分子分析

• 批准号: 10335237
• 负责人: Trisha N. Davis
• 金额: $ 73.08万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10335237
• 关键词: Address; Affinity; Aneuploidy; Avidity; Binding Proteins; Cell Death; Cell division; Chromosome Segregation; Chromosomes; Congenital Abnormality; DNA Binding; Exhibits; Genetic Materials; Geometry; Individual; Kinetochores; Lead; Life; Longevity; Measurement; Microtubules; Mitosis; Mitotic spindle; Molecular Analysis; Molecular Machines; Morphogenesis; Organelles; Ploidies; Process; Property; Proteins; Signal Transduction; Sum; Testing; Work; base; chromosome conformation capture; daughter cell; disability; in vivo; reconstitution; repaired; response; synergism; transmission process; tumorigenesis

Project Summary Life depends on the accurate transmission of genetic material at each cell division. Errors in this process lead to aneuploidy, which is implicated in oncogenesis, birth defects and cell death. Duplicated chromosomes are captured and segregated by a microtubule-based molecular machine, the mitotic spindle. The spindle is bipolar and each spindle pole carries an exact complement of chromosomes to each daughter cell. During mitosis, microtubules nucleate from the poles and capture and organize the chromosomes. Kinetochores, large multiprotein organelles located at the centromeric DNA, bind the microtubules and anchor the chromosomes to the poles. Our work focuses on each end of the microtubule, the spindle poles and the kinetochores. Spindle morphogenesis requires spatially controlled microtubule nucleation. Using a combination of reconstitution and in vivo analysis, we will test hypotheses that address how microtubule nucleation is activated and spatially regulated. Kinetochores attach chromosomes to microtubules with a striking combination of strength and plasticity. The attachments are mobile and robust under tension, but can also rapidly destabilize in response to regulatory signals. As such, the kinetochore is at the center of an error correction mechanism that repairs incorrect attachments sensed by a lack of ‘proper’ tension. The identification of the proteins that are under tension, the measurement of the strength of the linkages and the requirements for the full strength of attachments are together the second focus of this project. We have found that individually no kinetochore protein binds the microtubule with strength or longevity. To reconstitute the full strength of microtubule attachment exhibited by native kinetochores requires synergy between proteins in contact with the microtubule with proteins within the interior of the kinetochore. We will use a reconstitution-based approach and in vivo analysis to test the contribution of affinity, avidity and geometry to this synergy. In this way we will understand how the whole achieves greater properties than the sum of the parts. In addition, by exploiting our reconstituted kinetochore, we will test hypotheses for how the tension signal that triggers error correction is transmitted from the kinetochore and received by the repair mechanisms.

项目摘要 生命取决于在此过程中遗传材料的准确传播。 导致非整倍性，这与肿瘤发生，出生缺陷和细胞死亡有关。 染色体被基于微管的分子机捕获和分离有丝分裂 主轴。 每个女儿细胞。 染色体。 微管并将染色体锚定在极点。 微管，主轴杆和kinetchores。 纺锤形态发生需要空间控制的微管核。 重新结构和体内分析，我们将测试假设的假设，以解决微管核的含义。 激活并在空间上进行了规则。 Kinetchores将染色体连接到微管上，具有强度和可塑性的惊人组合。 在紧张局势下，Atachments是可移动和健壮的 监管信号。 缺乏“适当”的蛋白质，不正确 在紧张局势下，链接的测量和对全力的要求 atchments是该项目的第二个重点。 我们发现单个NO Kinetchore蛋白可以以强度或寿命结合微管。 重建本机Kinetchores展示的微管附件的全部强度 与微管接触的蛋白质之间的协同作用与蛋白质内部内部的蛋白质 Kinetchore。 与协同作用的亲和力，亲和力和几何形状。 除了利用我们的侦察吗？ 将测试假设的张力信号如何触发从您传输的误差校正 Kinetchore并通过维修机制接收。