Understanding developmental control of cell polarity using single-cell in vivo biochemistry

使用单细胞体内生物化学了解细胞极性的发育控制

基本信息

批准号：
10029493
负责人：
Daniel J Dickinson
金额：
$ 37.18万
依托单位：
UNIVERSITY OF TEXAS AT AUSTIN
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10029493
关键词：
Age Animals Anterior Apical Biochemical Biochemistry Biological Models Biological Process Caenorhabditis elegans Cell Cycle Cell Polarity Cell Separation Cells Characteristics Complex Cues Data Defect Development Disease Disseminated Malignant Neoplasm Embryo Eukaryotic Cell Foundations Genetic Goals Homeostasis In Vitro Individual Knowledge Learning Link Malignant Neoplasms Maps Measurement Membrane Methods Microscopy Mission Molecular Mutagenesis National Institute of General Medical Sciences Outcome Phosphotransferases Play Positioning Attribute Prevention Protein Kinase Proteins Regulation Research Role Signal Pathway Signal Transduction Somatic Cell Specific qualifier value Stereotyping System Testing Tissues Work age related base cell behavior cell type disease diagnosis experimental study extracellular in vivo in vivo Model innovation novel polarized cell prevent programs protein complex protein protein interaction response single molecule sperm cell tool zygote

项目摘要

Project Summary / Abstract Cell polarity is a fundamental feature of eukaryotic cells, and must be coordinated between cells and regulated to allow for normal animal development and tissue homeostasis. Despite genetic identification of proteins involved in cell polarity and a large body of knowledge about their interactions in vitro, it remains unclear how polarity proteins are organized into signaling complexes in cells. This lack of knowledge has prevented the field from understanding mechanisms of developmental control of polarity signaling in vivo. The long-term goal of the proposed research is to resolve the network of protein-protein interactions that supports animal cell polarity and to understand how this network can respond to developmental signals. To enable progress towards this goal, the applicants have developed innovative experimental tools that allow single-molecule measurements of native protein complex abundance in single cells. This project focuses on two evolutionarily conserved protein kinases, called aPKC and PAR-1, that play central roles in polarity by localizing to opposite ends of a polarized cell and dictating polarized cell behaviors. The applicants will make use of the C. elegans early embryo, in which cells reproducibly polarize in response to multiple spatial and temporal cues, to discover mechanistic links between developmental signals and the polarity machinery. The central hypothesis of this work is that that developmental signals control cell polarity by altering the molecular complexes in which aPKC and PAR-1 reside. This hypothesis will be explored by identifying dynamic aPKC and PAR-1 complexes that control polarity (Aim 1); by determining how polarity signaling is coordinated with cell cycle cues in the zygote (Aim 2); and by determining how developmental cues re-program polarity signaling in later embryos. The work proposed in this application is significant because it will reveal fundamental mechanisms controlling cell polarity, and because it places these mechanistic studies in a developmental context. The proposed work is innovative, in the applicant’s opinion, because it uses novel experimental methods to perform biochemical, mechanistic studies in vivo. By studying the biochemical control of aPKC and PAR-1 in multiple cellular and developmental contexts in a single experimental system, this work will identify fundamental mechanisms of PAR polarity signaling and to learn how these mechanisms are deployed to achieve different outcomes during development.

项目概要/摘要细胞极性是真核细胞的基本特征，必须在细胞之间协调和调节尽管对蛋白质进行了遗传鉴定，但仍能实现正常的动物发育和组织稳态。参与细胞极性以及关于它们在体外相互作用的大量知识，目前尚不清楚如何极性蛋白在细胞中被组织成信号复合物，这种知识的缺乏阻碍了这一领域的发展。来自理解体内极性信号发育控制的机制。拟议研究的长期目标是解决蛋白质-蛋白质相互作用的网络支持动物细胞极性并了解该网络如何响应发育信号。为了实现这一目标，申请人开发了创新的实验工具，允许该项目重点关注单细胞中天然蛋白质复合物丰度的单分子测量。两种进化上保守的蛋白激酶，称为 aPKC 和 PAR-1，通过以下方式在极性中发挥核心作用：定位到极化细胞的两端并指示极化细胞的行为。使用秀丽隐杆线虫早期胚胎，其中细胞响应多个空间和时间线索，发现发育信号和极性机制之间的机制联系。这项工作的中心假设是发育信号通过改变细胞极性来控制 aPKC 和 PAR-1 所在的分子复合物将通过鉴定来探索。通过确定极性信号如何控制极性的动态 aPKC 和 PAR-1 复合物；与受精卵中的细胞周期线索相协调（目标 2）并确定发育线索如何重新编程；本申请中提出的工作意义重大，因为它将揭示胚胎中的极性信号。控制细胞极性的基本机制，并且因为它将这些机制研究置于一个申请人认为，拟议的工作具有创新性，因为它使用了新颖的内容。通过研究生化控制进行体内生化、机制研究的实验方法。这项工作在单个实验系统中在多个细胞和发育环境中研究 aPKC 和 PAR-1 将识别 PAR 极性信号传导的基本机制并了解这些机制是如何运作的部署以在开发过程中实现不同的结果。