Class I myosins regulate protrusive forces at the actin-membrane interface

I 类肌球蛋白调节肌动蛋白-膜界面的突出力

基本信息

批准号：
10445354
负责人：
Mira Krendel
金额：
$ 53.4万
依托单位：
UPSTATE MEDICAL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-06 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10445354
关键词：
Actins Adhesions Affect Automobile Driving Binding Biological Models Biological Process Cell Shape Cell membrane Cell model Cells Complex Cytoskeleton Data Endocytic Vesicle Endocytosis Environment Eukaryota Experimental Models Filament Fission Yeast Generations Immune response Knockout Mice Light Mammalian Cell Measures Mediating Membrane Microfilaments Modeling Molecular Molecular Motors Motor Motor Activity Mus Mutation Myosin ATPase Myosin Type I Nonmuscle Myosin Type IIA Nutrient Phagocytes Phagocytosis Physiological Play Plus End of the Actin Filament Proteins Regulation Regulation of Cell Shape Resistance Role Shapes Site Structure System Tail Techniques Testing Work Yeast Model System Yeasts base cell growth cell motility experimental study extracellular macrophage migration monomer mutant pathogen polymerization pressure protein crosslink recruit

项目摘要

In this project, we will dissect the roles of class I myosins, molecular motors that bind actin filaments through their motor domains and plasma membrane through their tail domains, in actin-mediated cell shape changes. We will test the hypothesis that myosin I motor activity promotes assembly of branched actin networks at the sites of membrane deformation, including endocytic vesicles, phagocytic cups, and podosomes. The proposed work will use two model systems: fission yeast, where myosin I is required for endocytosis, and murine macrophages, which rely on myosin I for phagocytosis and cell migration. In aim 1, we will determine how myosin motor activity contributes to endocytic actin patch assembly and internalization in yeast. We will use the genetically tractable yeast system to express mutant myosin from the endogenous myosin locus and apply molecule counting techniques, which are well established in the yeast endocytosis system, to measure the rates of actin assembly. In aim 2, we will test the role of myosin motor activity in actin and membrane dynamics during phagocytosis and cell migration. These studies will take advantage of the genetically modified mice lacking class I myosins that are available in our labs. In aim 3, we will determine how myosin-membrane interactions are regulated to contribute to myosin functions in actin assembly and immune response. The proposed work will shed light on the molecular mechanisms driving actin- dependent membrane deformation.

在这个项目中，我们将剖析 I 类肌球蛋白（结合肌动蛋白的分子马达）的作用细丝通过其运动域，质膜通过其尾部域，肌动蛋白介导的细胞形状变化。我们将检验肌球蛋白 I 运动活动的假设促进膜变形部位分支肌动蛋白网络的组装，包括内吞囊泡、吞噬杯和足小体。建议的工作将使用两种模型系统：裂殖酵母（内吞作用需要肌球蛋白 I）和鼠巨噬细胞，依赖肌球蛋白 I 进行吞噬作用和细胞迁移。在目标 1 中，我们将确定如何肌球蛋白运动活性有助于酵母内吞肌动蛋白补丁的组装和内化。我们将使用遗传易处理的酵母系统来表达来自内源性肌球蛋白位点并应用分子计数技术，该技术已得到广泛应用在酵母内吞系统中，测量肌动蛋白组装的速率。在目标 2 中，我们将测试吞噬作用和细胞过程中肌球蛋白运动活性在肌动蛋白和膜动力学中的作用迁移。这些研究将利用缺乏 I 类基因的转基因小鼠我们实验室提供的肌球蛋白。在目标 3 中，我们将确定肌球蛋白膜如何相互作用被调节以促进肌动蛋白组装和免疫中的肌球蛋白功能回复。拟议的工作将揭示驱动肌动蛋白的分子机制依赖膜变形。