Assembly and function of cytoskeletal systems in eukaryotic and prokaryotic cells

真核和原核细胞中细胞骨架系统的组装和功能

基本信息

批准号：
9900836
负责人：
R DYCHE MULLINS
金额：
$ 47.55万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
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：
2016-04-15 至 2021-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9900836
关键词：
3-Dimensional Actins Archaea Architecture Biochemical Biological Biological Process Biophysical Process Biophysics Cell Nucleus Cells Cellular Structures Cellular biology Collection Complex Cytoskeleton DNA DNA Damage DNA Double Strand Break Endocytosis Eukaryota Eukaryotic Cell Family Filament Funding Genome Goals Grant In Vitro Individual Laboratories Life Membrane Microfilaments Microscopic Microscopy Molecular Motor Movement National Institute of General Medical Sciences Nuclear Phagocytosis Polymers Prokaryotic Cells Property Protein Family Proteins Resolution Role Rupture Shapes Signal Pathway Structure Work base cell motility cell type cofilin comparative genomics healing macromolecule mechanical force migration pathogen profilin public health relevance reconstitution response segregation self assembly single molecule vasodilator-stimulated phosphoprotein

项目摘要

DESCRIPTION (provided by applicant): A major focus of my laboratory is to understand how cytoskeletal polymers help a collection of macromolecules work together to establish a common identity: to become a living cell. Grants to my laboratory from NIGMS have funded work on several basic cell biological processes, in both eukaryotes and prokaryotes, including: (i) the assembly and function of force-generating `lamellipodial' actin networks that drive membrane movements in eukaryotic cells (GM061010 and an NDC Roadmap Grant); (ii) the actin nucleating activity and biological function of Spire-family proteins (GM075287); (iii) the architecture and function of actin-based structures in the nucleus (GM061010); and (iv) DNA segregation in bacterial cells, driven by assembly of cytoskeletal polymers (GM095263 and GM079556). A MIRA grant funding all these projects would enable us redirect energy previously devoted to maintaining multiple R0-1 grants into doing more original work. To understand how molecular properties govern the architecture and function of living cells, we perform quantitative studies at multiple size scales: (i) single-molecule and bulk biochemical studies of cytoskeletal components; (ii) biophysical and microscopical studies of complex cellular structures reconstituted in vitro; and (iii) cell biological and high-resolution microscopy studies of cytoskeletal systems in living cells. Broadly speaking, the ongoing work that would be supported by this grant can be divided into four parts: 1. Studies of prokaryotic cytoskeletal systems. Prokaryotic genomes encode more than forty classes of actin-like proteins (ALPs). We have studied the assembly and function of three eubacterial ALPs (ParM, AlfA, and Alp7A) and we are currently working to understand the role of ALPs in archaea. 2. Studies of dendritic actin networks assembled by: the Arp2/3 complex, WASP/WAVE-family proteins, Ena/VASP, formins, capping protein, cofilin, and profilin. In addition to powering some types of cell locomotion, this `dendritic network motor' also contributes to phagocytosis, endocytosis, movement of intracellular pathogens, and healing of membrane ruptures. Among other things, we are currently working to understand the mechanisms by which these networks generate and adapt to mechanical forces. 3. Studies of fast amoeboid migration. We combine `Evolutionary Cell Biology' approaches with 3D "Bessel Beam" microscopy to understand how cells generate complex membrane dynamics and then harness them for rapid movement. Briefly, we employ comparative genomics and cell biological studies of widely divergent (non-model) organisms to uncover molecular and biophysical mechanisms of cell movement. 4. Studies of the assembly and function of actin filaments in eukaryotic nuclei. We recently discovered nuclear actin filaments created by Fmn2 and Spire-family molecules in response to DNA damage. These filaments contribute to rapid clearance of double-strand DNA breaks (Belin and Mullins, submitted) and we are working to understand their functions and to work out the signaling pathways that create them.

描述（由适用提供）：我的实验室的主要重点是了解细胞骨架聚合物如何帮助一系列大分子共同工作以建立共同的身份：成为活细胞。通过Nigms向我的实验室赠款，已经为真核生物和原核生物的几个基本细胞生物过程提供了资金，包括：（i）生成力的“ lamellipodial”肌动蛋白网络的组装和功能，这些肌动蛋白网络可驱动真核生物细胞中的膜运动（GM06101010和NDC Roadmap Roadmap Grans）；（ii）尖峰家庭蛋白的肌动蛋白成核活性和生物学功能（GM075287）；（iii）基于肌动蛋白的结构在细胞核中的结构和功能（GM061010）；（iv）细菌细胞中的DNA分离，由细胞骨架聚合物组装（GM095263和GM079556）驱动。 MIRA赠款资助所有这些项目将使我们能够重定向以前致力于维持多个R0-1赠款的能源，以进行更多的原创工作。为了了解分子特性如何控制活细胞的结构和功能，我们在多个尺寸尺度上进行定量研究：（i）细胞骨架成分的单分子和大量生化研究；（ii）在体外重构的复杂细胞结构的生物物理和微观研究；（iii）活细胞中细胞骨架系统的细胞生物学和高分辨率显微镜研究。从广义上讲，这笔赠款将支持的正在进行的工作可以分为四个部分：1。核核骨骼系统的研究。原核基因组编码了40种肌动蛋白样蛋白（ALP）。我们研究了三个Eubacterial Alps（Parm，Alfa和Alp7a）的组装和功能，并且我们目前正在努力了解阿尔卑斯山在古细菌中的作用。 2。组装的树突状肌动蛋白网络的研究：ARP2/3复合物，WASP/波型蛋白质，ENA/VASP，formins，formins，Capping蛋白，cofilin和profilin。除了为某些类型的细胞运动提供动力外， “树突网络运动”也有助于吞噬作用，内吞作用，细胞内病原体的运动以及膜破裂的愈合。除其他外，我们目前正在努力了解这些网络生成并适应机械力的机制。 3。快速迁移的研究。我们将“进化细胞生物学”方法与3D“贝塞尔束”显微镜结合在一起，以了解细胞如何产生复杂的膜动力学，然后利用它们进行快速运动。简而言之，我们采用了比较基因组学和细胞生物学研究，对广泛不同（非模型）生物的生物进行了细胞运动的分子和生物物理机制。 4。对真核核中肌动蛋白丝的组装和功能的研究。我们最近发现了FMN2和尖峰家庭分子对DNA损伤产生的核肌动蛋白丝。这些细丝有助于快速清除双链DNA断裂（Belin和Mullins，已提交），我们正在努力理解它们的功能并确定创建它们的信号传导途径。