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.

项目总结/摘要： 该项目为申请人提供了博士学位。分子和细胞生物学及生物化学方面的培训。的 申请人的论文研究将进一步我们的细胞极化是如何调节过程中的理解 多细胞生物的发展。细胞极性，或细胞成分的组织， 在细胞的不同区域，是不同细胞过程的基础，如不对称细胞分裂和细胞分裂。 迁移细胞极性的核心是进化上保守的Par复合物，它指导亚细胞的极性。 通过磷酸化定位下游因子。Par复合物磷酸化了大量的 下游因素，但只有在严格的空间和时间要求得到满足。虽然执行 这些时空要求是正确的Par复函数的关键，但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Cdc 42如何协同工作来介导Par 复杂函数Par复合物在细胞皮层的亚细胞定位由Par-3调节， 抑制Par复合物活性，因为它不对称地将其靶向皮层。一旦适当定位，Par复合体 从Par-3转变为Cdc 42，刺激Par复合物活性。因此，从Par-3到Cdc 42的转变是 对Par复合物活性调节至关重要。虽然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，详细说明Par复合体如何从非活性Par-3- 与活性Cdc 42结合复合物结合的问题仍未解决。此外，这些复合物的活性具有 Par-3或Cdc 42如何调节Par复合物活性的机制也未被直接研究。 申请人的初步生物化学数据表明，Par-3和Cdc 42抑制彼此与CD 42的结合。 标准杆复合体。申请人将解决我们对Par理解中的几个基本知识差距 介导的极性使用广泛的生物化学技术。申请人将决定哪一个利息和 分子内相互作用控制Par复合物从Par-3转换为Cdc 42的能力（目的1），研究 Par复合物转换如何通过检查细胞类型特异性Par的作用来调节不同的细胞类型。 复合物调节剂Crumbs（目的2），并检验Par-3结合的Par复合物催化活性较低的假设 活性高于Cdc 42-或Crumbs-结合的Par复合物（Aim 3）。总之，拟议的目标将提供一个全面的 了解Par复合物的定位和活性在不同细胞中的调节机制， contexts.描述这一关键的调节轴将提供对发育缺陷和疾病的洞察力 其与PAR介导的极性功能障碍有关。申请人将获得广泛的技术 由经验丰富的导师团队指导的培训。此外，该奖学金将为申请人提供 科学交流、教育和专业发展机会。