CAREER: Mechanisms controlling spatial patterning of PIP lipids in eukaryotic cell polarity

职业：真核细胞极性中 PIP 脂质空间模式的控制机制

• 批准号: 2048060
• 负责人: Scott Hansen
• 金额: $ 103.92万
• 依托单位: University of Oregon Eugene
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2026-01-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2048060&HistoricalAwards=false
• 关键词: CAREER; Mechanisms; controlling; spatial; patterning

The ability of cells to regulate the localization of molecules in both time and space is a hallmark of cellular organization. In eukaryotic cells, phosphatidylinositol phosphate (PIP) lipids function as master regulators of cellular organization by selectively recruiting proteins to intracellular membranes and locally controlling their activity. Using a multidisciplinary biochemistry approach, this project will help decipher mechanisms that regulate communication between signaling molecules that control cellular organization. In long-term, this project will provide a molecular understanding of tissue organization, asymmetric cell division, and cell polarity. Integrated with this research strategy, the education plan seeks to improve how early career scientists communicate their research using art and scientific imagery as a tool for promoting inquiry-based learning. Undergraduate researchers will receive training in visual arts to improve their ability to communicate science. The education plan will support career development in scientific illustration and molecular visualization that bridges science, art, and design.The goal of the proposed research is to determine how biochemical reactions spontaneously self-organize on the plasma membrane to drive cell polarity. To achieve this goal, this project will use a bottom-up approach to biochemically reconstitute the communication between several important signaling elements, including (1) phosphatidylinositol phosphate (PIP) lipids, (2) Rho-GTPases, and (3) the actin cytoskeleton on membrane surfaces. Measurements, implemented in this project, will span several length scales beginning with single molecule biophysical measurements of PIP lipid modifying enzymes on supported membranes using Total Internal Reflection Fluorescence (TIRF) microscopy. Individual biochemical reactions will be combined to generate reaction diffusion systems that exhibit emergent properties, including bistability and polarization on membranes. This research will establish methods to directly visualize interactions between polarized signaling domains and the actin cytoskeleton using micropatterned supported bilayers and membrane encapsulation. Deterministic and stochastic kinetic modeling will be used to create a theoretical framework that describes the enzymology and spatial patterning mechanisms investigated. Overall, the proposed research will identify principles and signaling network architectures that are important for establishing communication between different classes of membrane signaling reactions that control cell polarity. This project is supported by the Molecular Biophysics and Cellular Dynamics and Function Clusters of the Molecular and Cellular Biosciences Division in the Directorate for Biological Sciences.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

细胞在时间和空间上调节分子定位的能力是细胞组织的标志。在真核细胞中，磷脂酰肌醇磷酸（PIP）脂质通过选择性地将蛋白质募集到细胞内膜并局部控制其活性来作为细胞组织的主要调节剂。使用多学科的生物化学方法，该项目将有助于破译调节控制细胞组织的信号分子之间的通信机制。从长远来看，该项目将提供对组织组织，不对称细胞分裂和细胞极性的分子理解。与这一研究策略相结合，该教育计划旨在改善早期职业科学家如何利用艺术和科学图像作为促进探究式学习的工具来传播他们的研究。本科研究人员将接受视觉艺术培训，以提高他们沟通科学的能力。该教育计划将支持科学插图和分子可视化的职业发展，连接科学，艺术和设计。拟议研究的目标是确定生物化学反应如何在质膜上自发自组织以驱动细胞极性。为了实现这一目标，该项目将使用自下而上的方法来生化重建几个重要信号元件之间的通信，包括（1）磷脂酰肌醇磷酸（PIP）脂质，（2）Rho-GTP酶，和（3）膜表面的肌动蛋白细胞骨架。 测量，在这个项目中实施，将跨越几个长度尺度开始与单分子生物物理测量PIP脂质修饰酶的支持膜使用全内反射荧光（TIRF）显微镜。单个的生化反应将被结合起来，以产生反应扩散系统，表现出新兴的属性，包括双稳态和膜上的极化。本研究将建立方法，直接可视化极化信号结构域和肌动蛋白细胞骨架之间的相互作用，使用微图案化的支持双层和膜封装。确定性和随机动力学建模将被用来创建一个理论框架，描述了酶学和空间模式机制的调查。总的来说，拟议的研究将确定原则和信号网络架构，这些原则和信号网络架构对于在控制细胞极性的不同类型的膜信号反应之间建立通信非常重要。该项目由生物科学理事会分子和细胞生物科学部的分子生物物理学和细胞动力学及功能群支持。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

A novel homeostatic mechanism tunes PI(4,5)P2-dependent signaling at the plasma membrane.

• DOI: 10.1242/jcs.261494
• 发表时间: 2023-08-15
• 期刊: Journal of cell science
• 影响因子: 4