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; 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了博士学位。分子和细胞生物学和生物化学的培训。这 申请人的论文研究将进一步了解我们对细胞极化如何调节的理解 多细胞生物的发展。细胞极性，或组织特定的细胞成分 细胞的区域对于多种细胞过程，例如不对称细胞分裂和细胞是基础 迁移。细胞极性的核心是进化保守的PAR复合物，它指导亚细胞 通过磷酸化来定位下游因子。 PAR复合物磷酸化众多 下游因素，但只有满足严格的空间和临时要求。而执行 这些时空要求是适当的PAR复杂功能的关键，该领域并不完全理解这一点 过程。该项目将研究两种调节蛋白（Par-3和Cdc42）如何协同工作以调节PAR 复杂函数。 PAR在细胞皮层处的PAR复合物的亚细胞定位受PAR-3的调节，这 抑制PAR复杂活性，因为它不对称地将其靶向皮层。一旦适当定位，par综合体 从par-3到Cdc42的过渡，刺激了PAR复杂活性。那就是从第3杆到cdc42的过渡 对于PAR复杂活性调节至关重要。尽管这种过渡是当前细胞模型的主要特征 潜水细胞类型中的极性，该机制详细介绍了PAR复合物如何从非活性Par-3-- 与活动CDC42结合的复合物结合保持未解决。另外，这些复合物的活性具有 未直接检查了PAR-3或CDC42如何调节PAR复杂活性的机制。 申请人的初步生化数据表明，PAR-3和CDC42抑制了对方对彼此的结合 par复合体。申请人将解决我们对PAR的理解时的几个基本知识差距 使用多种生化技术介导的极性。申请人将解决哪些与 分子内相互作用控制着PAR复合物从PAR-3转换为Cdc42的能力（AIM 1），调查 PAR复杂开关如何通过检查细胞类型特异性PAR的作用来调节潜水细胞类型 复杂的调节剂，面包屑（AIM 2），并检验了par-3结合PAR复合物的假设较小的催化剂 比Cdc42-或面包屑结合的PAR复合物活跃（AIM 3）。拟议的目标在一起将提供直接的 机械理解如何在不同的细胞中调节PAR复合物的定位和活性 上下文。表征这种关键的调节轴将为发育缺陷和疾病提供见解 与PAR介导的极性功能障碍相关的。申请人将获得广泛的技术 由经验丰富的训练团队指导的培训。此外，该奖学金将为申请人提供 科学沟通，教育和专业发展机会。