Probing Actin Filament Assembly, Structure, and Dynamics by Palladin

Palladin 探索肌动蛋白丝组装、结构和动力学

基本信息

批准号：
9171290
负责人：
MORIAH R BECK
金额：
$ 41.53万
依托单位：
WICHITA STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-01 至 2021-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9171290
关键词：
Actin-Binding Protein Actins Address Affect Architecture Automobile Driving Behavior Binding Biochemical Biology Cardiomyopathies Cell Nucleus Cell division Cell physiology Cells Characteristics Complement Complex Coupling Cytoskeleton Data Development Disease Disseminated Malignant Neoplasm Electrons Embryonic Development F-Actin Filament Fluorescence Microscopy G Actin Goals Imaging Techniques Kinetics Lead Life Listeria Listeria monocytogenes Mediating Microfilaments Modeling Molecular Monitor Neoplasm Metastasis Normal Cell Pharmaceutical Preparations Play Population Process Proteins Regulation Research Role Staging Structure Tail Testing Time Work Wound Healing base cancer cell cell motility crosslink depolymerization in vitro Assay in vivo insight mammalian embryology neoplastic cell network architecture overexpression pathogen polymerization research study therapeutic target three dimensional structure tumor progression

项目摘要

PROJECT SUMMARY/ABSTRACT Remodeling of actin filaments is essential for normal cell processes such as wound healing, cell division, and embryonic development. However alterations in the actin cytoskeleton are also associated with serious diseases such as metastatic cancer and cardiomyopathy. Understanding the fundamental mechanisms that govern actin-based cell motility is an important unsolved problem in biology that is critical for developing therapeutics that target alterations in actin dynamics and organization. In order to understand the molecular mechanisms that regulate the assembly of actin filaments, we are focusing on palladin, which we have recently shown can enhance actin polymerization. Normal embryonic development requires palladin and overexpression of palladin is correlated with invasive motility associated with metastatic cancer, however the precise role of palladin in cell motility has yet to be determined. Our current model suggests that palladin promotes actin nucleation and stabilizes actin filaments and that these functions are critical for regulating actin dynamics and organization within the cell. In the proposed studies we will rigorously test this model, determining whether palladin is key for actin-based cell motility and exploring the critical interactions involved to unravel the mechanism. In Aim 1 of this proposal, we will analyze the role of palladin in nucleation of G-actin by testing different nucleation mechanisms. In Aim 2, we will determine the effect of palladin-induced polymerization on actin network architecture. In Aim 3, we will complement our quantitative assays of in vitro actin kinetics with in vivo monitoring of actin-based motility driving the formation of Listeria comet tails. This combination of experiments will allow us to address the question of how palladin contributes to actin-based actin dynamics and organization; and moreover how this role changes with increased expression of palladin.

项目摘要/摘要肌动蛋白丝的重塑对于正常细胞过程，例如伤口愈合，细胞至关重要分区和胚胎发展。但是，肌动蛋白细胞骨架的改变也是与严重疾病（如转移性癌症和心肌病）相关。了解控制基于肌动蛋白的细胞运动的基本机制是一种生物学中重要的未解决问题，这对于开发针对目标的治疗剂至关重要肌动蛋白动态和组织的改变。为了理解分子调节肌动蛋白丝组装的机制，我们专注于palladin，我们最近显示的可以增强肌动蛋白聚合。正常的胚胎发育需要Palladin和Palladin的过表达与与侵入性运动相关在转移性癌症中，palladin在细胞运动中的确切作用尚未是决定。我们当前的模型表明，palladin促进肌动蛋白成核并稳定肌动蛋白丝，这些功能对于调节肌动蛋白动态和细胞内的组织。在拟议的研究中，我们将严格测试此模型，确定palladin是否是基于肌动蛋白的细胞运动并探索关键的关键涉及阐明机制的相互作用。在本提案的目标1中，我们将分析通过测试不同的成核机制，帕拉丁素在G-肌动蛋白成核中的作用。在AIM 2中，我们将确定palladin诱导的聚合对肌动蛋白网络架构的影响。目标 3，我们将通过体内监测的体外肌动蛋白动力学的定量测定基于肌动蛋白的运动驱动李斯特菌彗星尾巴的形成。实验的组合将使我们能够解决Palladin如何对基于肌动蛋白的肌动蛋白动态做出贡献的问题和组织；此外，这种作用如何随palladin的表达增加而变化。