Elucidating the mechanism of cell type specific regulation of the Par complex

阐明 Par 复合物的细胞类型特异性调节机制

• 批准号: 10607140
• 负责人: Elizabeth Vargas
• 金额: $ 3.91万
• 依托单位: UNIVERSITY OF OREGON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10607140
• 关键词: Adaptor Signaling Protein; Affect; Allosteric Regulation; Animals; Binding; Biochemical; Biochemistry and Cellular Biology; Biological Assay; Cell Extracts; Cell Polarity; Cell division; Cell model; Cell physiology; Cells; Cellularity; Communication; Complex; Cues; Data; Defect; Development; Disease; Doctor of Philosophy; Education; Elements; Epithelial Cells; Epithelium; Exclusion; Fellowship; Foundations; Functional disorder; Future; Health; In Vitro; Knowledge; Length; Link; Mediating; Mediator; Mentorship; Modeling; Molecular and Cellular Biology; Nature; Organism; PAR-6 protein; Phosphorylation; Phosphotransferases; Process; Protein Truncation; Proteins; Radiolabeled; Regulation; Reporting; Repression; Research; Role; Science; Site; Structure; Switching Complex; System; Techniques; Testing; Training; Work; cell cortex; cell motility; cell type; experience; experimental study; genetic regulatory protein; in vivo; innovation; insight; novel therapeutics; polarized cell; predictive modeling; reconstitution; spatiotemporal; tumorigenesis

PROJECT SUMMARY/ ABSTRACT: This project provides the applicant with Ph.D. training in molecular and cellular biology and biochemistry. The applicant’s thesis research will further our understanding of how cell polarization is regulated during the development of multicellular organisms. Cell polarity, or the organization of cellular components to specific regions of the cell, is fundamental for diverse cellular processes such as asymmetric cell division and cell migration. At the core of cell polarity is the evolutionarily conserved Par complex, which guides the subcellular localization of downstream factors by phosphorylation. The Par complex phosphorylates a multitude of downstream factors, but only if strict spatial and temporal requirements are satisfied. While the enforcement of these spatiotemporal requirements is key to proper Par complex function, the field does not fully understand this process. This project will examine how two regulatory proteins, Par-3 and Cdc42, work in concert to mediate Par complex function. Subcellular localization of the Par complex at the cell cortex is regulated by Par-3, which inhibits Par complex activity as it asymmetrically targets it to the cortex. Once properly localized, the Par complex transitions from Par-3 to Cdc42, stimulating Par complex activity. Thus, the transition from Par-3 to Cdc42 is crucial for Par complex activity regulation. Although this transition is a central feature of current models of cell polarity in diverse cell types, the mechanism detailing how the Par complex transitions from an inactive Par-3- bound to an active Cdc42-bound complex remains unresolved. Additionally, the activity of these complexes has not been directly examined nor is the mechanism known for how Par-3 or Cdc42 regulate Par complex activity. The applicant’s preliminary biochemical data indicates that Par-3 and Cdc42 inhibit each other’s binding to the Par complex. The applicant will address several fundamental knowledge gaps in our understanding of Par mediated polarity using a wide range of biochemical techniques. The applicant will resolve which inter and intramolecular interactions control the Par complex’s ability to switch from a Par-3 to Cdc42 (Aim 1), investigate how Par complex switching is regulated diverse cell types by examining the role of the cell type specific Par complex regulator, Crumbs (Aim 2), and test the hypothesis that Par-3-bound Par complex is less catalytically active than Cdc42- or Crumbs-bound Par complex (Aim 3). Together, the proposed aims will provide a thorough mechanistic understanding for how the Par complex localization and activity is regulated in different cellular contexts. Characterizing this critical regulatory axis will provide insight for developmental defects and disease which have been linked to Par-mediated polarity dysfunction. The applicant will receive extensive technical training guided by an experienced mentorship team. Additionally, this fellowship will provide the applicant with science communication, educational, and professional development opportunities.

项目摘要/摘要： 该项目为申请人提供博士学位。分子和细胞生物学和生物化学方面的培训。这 申请人的论文研究将进一步了解细胞极化在细胞极化过程中是如何调节的 多细胞生物的发育。细胞极性，或细胞成分的特定组织 细胞区域，对于不同的细胞过程（例如不对称细胞分裂和细胞分裂）至关重要。 迁移。细胞极性的核心是进化上保守的 Par 复合体，它指导亚细胞 通过磷酸化定位下游因子。 Par 复合物磷酸化大量 下游因素，但前提是满足严格的空间和时间要求。在执法的同时 这些时空要求是正确 Par 复杂函数的关键，该领域并未完全理解这一点 过程。该项目将研究两种调节蛋白 Par-3 和 Cdc42 如何协同作用来介导 Par 复杂的功能。 Par 复合物在细胞皮层的亚细胞定位受 Par-3 调节，Par-3 抑制 Par 复合体活性，因为它不对称地靶向皮质。一旦正确定位，Par 复合体 从 Par-3 过渡到 Cdc42，刺激 Par 复合体活动。因此，从 Par-3 到 Cdc42 的过渡是 对于 Par 复杂活动调节至关重要。尽管这种转变是当前细胞模型的核心特征 不同细胞类型中的极性，详细描述了 Par 复合物如何从非活性 Par-3- 转变的机制 与活性 Cdc42 结合复合物的结合仍未解决。此外，这些复合物的活性 Par-3 或 Cdc42 如何调节 Par 复合物活性的机制尚未被直接检查，也不清楚。 申请人的初步生化数据表明Par-3和Cdc42相互抑制与 帕复合体。申请人将解决我们对 Par 的理解中的几个基本知识差距 使用广泛的生化技术介导极性。申请人将决定哪些内部和 分子内相互作用控制 Par 复合物从 Par-3 切换到 Cdc42 的能力（目标 1），研究 通过检查细胞类型特异性 Par 的作用，如何调节不同细胞类型的 Par 复合物转换 复合物调节剂 Crumbs（目标 2），并测试 Par-3 结合的 Par 复合物催化作用较低的假设 比 Cdc42 或 Crumbs 结合的 Par 复合体更活跃（目标 3）。总之，拟议的目标将提供彻底的 对 Par 复合物定位和活性如何在不同细胞中进行调节的机制理解 上下文。表征这一关键的调控轴将为发育缺陷和疾病提供见解 这与 Par 介导的极性功能障碍有关。申请人将获得广泛的技术 培训由经验丰富的导师团队指导。此外，该奖学金将为申请人提供 科学传播、教育和专业发展机会。