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： 该项目为申请者提供分子、细胞生物学和生物化学方面的博士培训。这个 申请者的论文研究将进一步加深我们对细胞极化是如何在 多细胞生物体的发展。细胞极性，或细胞成分的组织以特定的 细胞的不同区域，是不同细胞过程的基础，如细胞不对称分裂和细胞 迁移。细胞极性的核心是进化上保守的PAR复合体，它引导亚细胞 通过磷酸化定位下游因子。PAR复合体磷酸化了大量的 下游因素，但只有在满足严格的空间和时间要求的情况下。在执法的同时， 这些时空要求是正确的PAR复合功能的关键，该领域并不完全理解这一点 进程。这个项目将研究两种调节蛋白，par-3和cdc42如何协同工作来调节par。 复杂的函数。PAR复合体在细胞皮质的亚细胞定位由PAR-3调节，它 抑制PAR复合体的活性，因为它不对称地将其靶向皮质。一旦正确本地化，PAR情结 从PAR-3到CDC42的转变，刺激PAR复合体的活性。因此，从PAR-3到CDC42的过渡是 对PAR复合体活性调节至关重要。尽管这种过渡是当前细胞模型的中心特征 不同细胞类型中的极性，详细说明了PAR复合体如何从不活跃的PAR-3- 结合到活性的CdC42结合的络合物仍然没有被分解。此外，这些络合物的活性具有 PAR-3或CDC42如何调节PAR复合体活性的机制尚未被直接研究。 申请人的初步生化数据表明，PAR-3和CDC42相互抑制彼此与 标准杆情结。申请者将解决我们对PAR理解中的几个基本知识差距 使用广泛的生化技术来调节极性。申请者将决定哪一家公司和 分子内相互作用控制PAR复合体从PAR-3转换到CDC42的能力(目标1)，调查 通过研究细胞类型特定的PAR的作用，PAR复合体转换如何调节不同的细胞类型 复合调节剂，Crumbs(目标2)，并检验PAR-3结合的PAR复合体催化作用较弱的假设 比与CdC42或Crumbs结合的PAR复合体更有效(目标3)。总之，提议的目标将提供一个彻底的 PAR复合体在不同细胞中定位和活性调节的机制理解 上下文。表征这一关键的调节轴将为发育缺陷和疾病提供洞察力 这与PAR介导的极性功能障碍有关。申请者将获得广泛的技术支持 培训由经验丰富的指导团队指导。此外，这项奖学金将为申请者提供 科学交流、教育和职业发展机会。