Characterization of a Filopodial Myosin

丝状肌球蛋白的表征

• 批准号: 10403958
• 负责人: Casey W Eddington
• 金额: $ 3.25万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10403958
• 关键词: Ablation; Actins; Address; Amoeba genus; Animal Model; Binding; Biochemical; Biological Assay; Biological Models; Biotin; Cell physiology; Cells; Complex; Data; Development; Dictyostelium; Dimerization; Distant; Environment; Evolution; Filopodia; Future; Goals; Immune; Immune response; In Vitro; Invaded; Knowledge; Label; Ligase; Malignant Neoplasms; Mammals; Mass Spectrum Analysis; Measures; Mediating; Membrane; Microfilaments; Molecular Motors; Motor; Motor Activity; Myosin ATPase; Neoplasm Metastasis; Neurons; Organism; Pathogenesis; Phenotype; Play; Process; Property; Proteins; Rod; Role; Site; Structure; Surveys; Thinness; Tissues; Work; axon guidance; base; cancer cell; cell motility; cell type; cohort; crosslink; experimental study; genetic manipulation; in vivo; insight; mutant; particle; polymerization; prevent; recruit; single molecule; social; wound healing

Project Summary/Abstract Filopodia play important roles in a variety of cellular processes including immune response, neuronal pathogenesis, and wound healing. Cells use filopodia to survey their environment and are typically upregulated in motile cells. Filopodia are thin, rod-like cellular protrusions composed of parallel bundles of actin and found in divergent organisms such as the social amoeba Dictyostelium and metazoans. The core mechanisms behind filopodia formation are conserved throughout metazoans and amoebozoans, yet the mechanism of filopodia initiation is still unknown. MyTH4-FERM (MF) myosins, the amoeboid Myosin 7 (DdMyo7) and metazoan Myosin 10 (Myo10), are essential for filopodia initiation. They are targeted to the membrane, form initiation foci, and are subsequently enriched in filopodia tips during elongation. Filopodia initiation is being studied in the simple model organism Dictyostelium because it has minimal protein redundancy, allows for efficient genetic manipulation, and biochemical quantities of material are easily obtained. The ultimate goal of the proposed work is to understand the role of MF myosins during filopodia initiation. To accomplish this, I will 1) define the motor properties of DdMyo7 using standard motility assays and 2) identify DdMyo7 interacting partners using an unbiased proximity ligase screen. The data generated by the proposed experiments will provide critical information needed to understand how a filopodia myosin drives initiation.

项目摘要/摘要 丝状虫在包括免疫反应，神经元在内的多种细胞过程中起重要作用 发病机理和伤口愈合。细胞使用丝状虫调查其环境，通常会上调 在运动细胞中。丝状虫是由肌动蛋白平行束组成的薄棒状细胞突起，发现 诸如社会变形虫的分歧生物和后生动物。背后的核心机制 丝状形成在整个后生动物和变形虫群中都保存，但丝状虫的机制 启动仍然未知。 神经4-肌（MF）肌球蛋白，变形虫肌球蛋白7（DDMYO7）和后生肌球蛋白10（myo10）是 对于丝状启动至关重要。它们针对膜，形成启动焦点，随后是 在伸长过程中富含丝状尖端。在简单模型生物体中研究了丝状启动 dictyostelium，因为它具有最小的蛋白质冗余，允许有效的遗传操作，并且 很容易获得生化量的材料。拟议工作的最终目标是了解 MF肌球蛋白在丝状启动过程中的作用。为此，我将1）定义 DDMYO7使用标准运动分析和2）使用无偏接近的DDMYO7相互作用伴侣识别DDMYO7 连接酶屏幕。提出的实验生成的数据将提供所需的关键信息 了解丝状肌球蛋白如何驱动启动。