Spindle Assembly Checkpoint Silencing

主轴装配检查点消音

• 批准号: 10456802
• 负责人: Nicholas Gene Brown
• 金额: $ 38.88万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10456802
• 关键词: Aneuploidy; Binding; Biological Assay; Cell Cycle; Cells; Chemicals; Chromosome Segregation; Complex; Crystallography; Cyclin B; Development; Electron Microscopy; Ensure; Enzyme Kinetics; Enzymes; Human; MXI1 gene; Malignant Neoplasms; Mediating; Mitotic; Mitotic Checkpoint; Molecular Machines; Multiprotein Complexes; PTTG1 gene; Process; Protein Engineering; Research; Signal Pathway; Sister Chromatid; System; Therapeutic; Time; Triad Acrylic Resin; Ubiquitin; anaphase-promoting complex; base; crosslink; design; innovation; novel; prevent; public health relevance; ubiquitin ligase

ABSTRACT The Spindle Assembly Checkpoint (SAC) is a signaling pathway responsible for the fidelity of chromosome segregation. Disruption of this process leads to catastrophic cellular consequences, such as aneuploidy and cancer. The primary effector of the SAC is the inhibitory complex known as Mitotic Checkpoint Complex (MCC). MCC is responsible for binding and inhibiting the 1.2 MDa ubiquitin ligase Anaphase-Promoting Complex/Cyclosome (APC). Once all of the sister chromatids achieve proper bipolar orientation, MCC is dismantled. This restores APC activity to trigger the ubiquitin-mediated proteasomal destruction of key mitotic regulators, e.g. Cyclin B and Securin, permitting mitotic exit. The mechanisms of release and disassembly of APC-bound MCC (BUBR1, MAD2, BUB3, and CDC20- an APC coactivator) by a triad of large multiprotein enzymes remain poorly understood. I aim to dissect this process using an innovative technological approach involving enzyme kinetics, chemical crosslinking, protein engineering, electron microscopy, NMR, crystallography, and cell-based assays. Information generated from the proposed research will have a long- lasting impact on the cell cycle field and may enable the development of novel cancer therapeutics.

抽象的 纺锤体装配检查点 (SAC) 是负责染色体保真度的信号通路 隔离。这一过程的破坏会导致灾难性的细胞后果，例如非整倍体和 癌症。 SAC 的主要效应器是称为有丝分裂检查点复合物的抑制复合物 （中冶集团）。 MCC 负责结合和抑制 1.2 MDa 泛素连接酶后期促进 复合体/环体（APC）。一旦所有姐妹染色单体达到正确的双极性方向，MCC 就会 拆除。这会恢复 APC 活性，触发泛素介导的关键有丝分裂的蛋白酶体破坏 监管机构，例如Cyclin B 和 Securin，允许有丝分裂退出。释放和分解的机制 由大型多蛋白三联体形成的 APC 结合 MCC（BUBR1、MAD2、BUB3 和 CDC20 - APC 共激活剂） 人们对酶的了解仍然知之甚少。我的目标是使用创新的技术方法来剖析这个过程 涉及酶动力学、化学交联、蛋白质工程、电子显微镜、核磁共振、 晶体学和基于细胞的测定。拟议研究产生的信息将具有长期的影响 对细胞周期领域产生持久影响，并可能促进新型癌症疗法的开发。