ACTIN CABLE FORMATION COORDINATED BY APC, FORMINS AND MICROTUBULES IN ANIMAL DEVELOPMENT

动物发育中由 APC、Formins 和微管协调的肌动蛋白线形成

基本信息

批准号：
9923672
负责人：
BROOKE M MCCARTNEY
金额：
$ 29.17万
依托单位：
CARNEGIE-MELLON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-07-01 至 2023-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9923672
关键词：
APC gene APC2 gene Actins Acute Lymphocytic Leukemia Address Adenomatous Polyposis Coli Adenomatous Polyposis Coli Protein Alleles Animal Model Animals Architecture Binding Biochemical Biological Models CLIP-170 gene Cell Nucleus Cell physiology Cells Cellular biology Collaborations Colorectal Cancer Complex Coupling Cytoskeleton Data Development Developmental Biology Disease Drosophila genus Electron Microscopy Elongation Factor Feedback Fluorescence Microscopy Genetic Goals Grant Growth Hodgkin Disease Human Image In Vitro Investigation Lead Length Link MLL gene Mediating Microfilaments Microtubule-Associated Proteins Microtubules Modeling Molecular Morphogenesis Morphology Mutation Nuclear Nurses Oogenesis Organism Ovary Pattern Peptide Elongation Factor 2 Phosphorylation Phosphotransferases Physiological Plus End of the Microtubule Positioning Attribute Property Proteins Regulation Research Research Personnel Role Science Structure Suppressor Mutations Testing Time Tumor Suppressor Proteins Work base cell cortex design experience in vivo insight light microscopy mutant premature protein complex reconstitution single molecule spatiotemporal time use tumorigenesis vasodilator-stimulated phosphoprotein

项目摘要

Project Summary The goal of this proposal is to define the mechanisms controlling dynamic assembly and function of cellular actin structures in a living, developing animal. Recent advances in single molecule in vitro TIRF microscopy have made major contributions to our understanding of the molecular mechanisms controlling actin and microtubule rearrangements. However, the field is lagging behind in the integration of these in vitro mechanisms into a physiological context by testing them in animal models. To fill this gap, we will use a combined `top-down' and `bottom-up' strategy, bringing together the genetic and live imaging power of Drosophila with quantitative in vitro reconstitution and single molecule analysis. Specifically, we focus on a group of interacting actin assembly factors, which includes Adenomatous polyposis coli proteins 1 and 2 (APC1 and APC2) and the formin Diaphanous (Dia), and how they work together to build a striking array of long actin cables in Drosophila nurse cells at a specific stage of oogenesis. Our central hypothesis is that APC1, APC2, and Dia each make a mechanistically distinct and essential contribution to actin cable formation, and that the proper spatial and temporal pattern of cable assembly arises from these proteins working in concert under tight regulation by interactions with microtubules and microtubule-associated proteins (e.g., EB1 and CLIP-170), and regulated by GSK3ß kinase. By directly visualizing these mechanisms at the single molecule level in vitro, and using this information to guide rigorous multi-faceted tests in living Drosophila ovaries, we will arrive at an unprecedented level of mechanistic understanding of actin assembly and microtubule-actin cross-talk in the living animal. The specific aims are: (1) Determine how actin cable arrays are assembled during oogenesis by tightly controlled multi-component mechanisms involving collaborative actin filament nucleation and elongation factors; and (2) Define the mechanisms by which microtubules trigger actin cable emergence and interact with growing cables to guide their morphogenesis and their contact with the nucleus.

项目摘要该提案的目的是定义控制细胞动态组装和功能的机制肌动蛋白在生物中发展的动物中的结构。单分子体外TIRF显微镜的最新进展已经为我们理解控制肌动蛋白的分子机制做出了重大贡献微管重排。但是，该领域在体外的整合中落后通过在动物模型中测试物理环境的机制。为了填补这个空白，我们将使用结合了“自上而下”和“自下而上”策略，将遗传和现场成像能力融合在一起果蝇具有定量的体外重构和单分子分析。具体来说，我们专注于相互作用的肌动蛋白组装因子，其中包括腺瘤性息肉病大肠杆菌1和2 （APC1和APC2）以及formin diaphanous（DIA），以及它们如何共同构建罢工阵列果蝇护士细胞中的长肌动蛋白电缆在特定的卵子发生阶段。我们的中心假设是 APC1，APC2和DIA对肌动蛋白电缆的形成做出机械不同的贡献，电缆组件的适当空间和临时模式是由这些蛋白质产生的通过与微管和微管相关蛋白的相互作用在严格调节下的音乐会（例如，EB1 和夹170），由GSK3ß激酶调节。通过直接在单个上可视化这些机制体外分子水平，并使用此信息指导活果蝇中的严格多面测试卵巢，我们将达到对肌动蛋白大会的前所未有的机械理解水平生物动物中的微管 - 肌动蛋白串扰。具体目的是：（1）确定肌动蛋白电缆阵列通过严格控制的多组分机制在卵子发生过程中组装在一起肌动蛋白丝核和伸长因子；（2）定义微管触发的机制肌动蛋白电缆的出现并与生长的电缆相互作用，以指导其形态发生及其接触核。