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Dynein-adaptor interaction mechanisms and malfunction at atomic resolution

原子分辨率下的动力蛋白-适配器相互作用机制和故障

基本信息

批准号：
10450161
负责人：
Beat Rolf Vogeli
金额：
$ 32.49万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-09 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10450161
关键词：
3-Dimensional Adaptor Signaling Protein Animal Model Animals Binding Biophysics C-terminal Caenorhabditis elegans Carrier Proteins Cell division Cells Chromosome Segregation Collaborations Complex Disease Dynein ATPase Elements Endosomes Ensure Family Glass Goals Golgi Apparatus Human In Vitro Investigation Kinetics Kinetochores Lead Light Link Mediating Methods Microtubules Minus End of the Microtubule Mitosis Mitotic Chromosome Molecular Motor Motor Activity Movement Multiprotein Complexes Mutation NMR Spectroscopy Nature Nerve Degeneration Neurodegenerative Disorders Nuclear Nuclear Magnetic Resonance Pathogenicity Play Point Mutation Positioning Attribute Protein Dynamics Proteins RNA Research Resolution Rod Roentgen Rays Role Running Secretory Vesicles Signal Transduction Site Specificity Spinal Muscular Atrophy Tail Techniques Testing Therapeutic Intervention Thermodynamics Vesicle alpha helix base biophysical techniques cell motility chromosome movement disease-causing mutation dynactin experimental study flexibility free behavior innovation insight link protein migration mutant nanoscale neuron loss predictive test recruit

项目摘要

Project Summary Neurodegenerative diseases are mostly incurable conditions that result in progressive degeneration or death of nerve cells. Prominent examples such as Spinal Muscular Atrophy (SMA) are caused by mutations in the cytoplasmic dynein multi-protein complex. Dynein 1 participates in a wide array of cellular activities, ranging from the cargo transport of proteins, RNA, and vesicles to nuclear migration and cell division. Processive movement of dynein along microtubules requires the binding to the essential co-activator dynactin and specific adaptor proteins, which recruit cargo and facilitate movement by forming a stable ternary complex with dynein and dynactin. Our understanding of these interactions is limited and many questions remain: How do cargo adaptors bring dynein and dynactin together? How does this lead to activation of the motor and the initiation of transport? Which elements determine the adaptor specificity? How do the disease-causing mutations modify these interactions? Importantly, it is becoming increasingly evident that binding to light intermediate chain of dynein 1 (`LIC1') is a common feature of functionally distinct adaptors. We will focus our investigations on the interaction between the C-terminal tail of the LIC1 with the adaptors Spindly, BICD2 and Hook3, which each have important but distinct functions in transport. These studies are complicated by the fact that the disordered nature of C- terminal LIC1 makes it difficult to obtain a comprehensive dynamic view at atomic resolution from X-ray or EM techniques. However, we will take the innovative approach of using nuclear magnetic resonance (NMR) as a nano-scale `magnifying glass' that pinpoints interaction sites within the proteins at atomic resolution. Compared to other methods, NMR is uniquely suited to study disordered proteins and to characterize highly dynamic interactions. We propose four aims: (1) Structural and dynamical characterization of free C-terminal dynein LIC1. (2) Characterization of dynein LIC1 interaction with Spindly, BicD2 and Hook3, and their binding competition. (3) Prediction and testing of effects of mutations on dynein LIC1 - adaptor interaction. (4) Structural & dynamic basis of interaction triangle between dynein LIC1, Spindly and ROD/Zw10/Zwilch complex. Our experiments are therefore expected to provide detailed molecular insight into how human dynein LIC1 engages with structurally diverse dynein adaptors. Through collaboration, we will extend our findings into animal models. Overall, this research is significant because the presence of pathogenic point mutations in dynein and adaptor proteins suggests that these insights will also help to understand the diseases caused by malfunction of the dynein-driven cargo transport and will offer therapeutic intervention targets.

项目摘要神经退行性疾病大多是无法治愈的疾病，会导致进行性变性或神经细胞死亡。脊髓性肌萎缩症(SMA)等突出例子是由细胞质动力蛋白多蛋白复合体的突变。动力蛋白1参与了广泛的细胞活动，从蛋白质、RNA和囊泡的货物运输到核迁移和细胞分裂。动力蛋白沿微管的行进运动需要结合到基本的共激活动力蛋白和特定的接头蛋白，它们招募货物并促进运动通过与动力蛋白和动力肌动蛋白形成稳定的三元络合物。我们对这些互动的理解是仍然存在许多有限的问题：货物适配器如何将动力蛋白和动力蛋白结合在一起？多么这是否会导致马达的激活和运输的启动？哪些因素决定了适配器专一性？致病突变是如何改变这些相互作用的？重要的是，它是越来越明显的是，与动力蛋白1的轻质中间链(LIC1)结合是一种常见的功能截然不同的适配器的特点。我们将重点调查两国之间的相互作用 LIC1的C-端尾，带有细长的适配器，BICD2和Hook3，它们各自具有重要的但在运输中具有独特的功能。这些研究之所以复杂化，是因为C-C的无序性质终端LIC1使从X射线获得原子分辨率的全面动态图像变得困难或EM技术。然而，我们将采取创新的方法，使用核磁共振 (核磁共振)是一个纳米尺度的‘放大镜’，可以精确定位蛋白质内部的原子相互作用部位。决议。与其他方法相比，核磁共振特别适合于研究无序蛋白质和描述高度动态的交互作用。我们提出了四个目标：(1)结构和动力游离C-末端动力蛋白LIC1的表征。(2)动力蛋白LIC1与LIC1相互作用的特征 Spindly、BicD2和Hook3以及它们的结合竞争。(3)效果的预测和检验动力蛋白LIC1-接头相互作用的突变。(4)相互作用三角的结构和动力学基础在动力蛋白LIC1、纤毛状和Rod/Zw10/ZWilch复合体之间。因此，我们的实验是可以期待的为人类动力蛋白LIC1如何与结构多样化的结合提供详细的分子洞察力动力蛋白适配器。通过合作，我们将把我们的发现扩展到动物模型中。总体而言，这这项研究意义重大，因为动力蛋白和适配器中存在致病点突变蛋白质表明，这些洞察力也将有助于理解由功能障碍引起的疾病动力蛋白驱动的货物运输，并将提供治疗干预目标。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The Inherent Dynamics and Interaction Sites of the SARS-CoV-2 Nucleocapsid N-Terminal Region.

DOI：
10.1016/j.jmb.2021.167108
发表时间：
2021-07-23
期刊：
Journal of molecular biology
影响因子：
5.6
作者：
Redzic JS;Lee E;Born A;Issaian A;Henen MA;Nichols PJ;Blue A;Hansen KC;D'Alessandro A;Vögeli B;Eisenmesser EZ
通讯作者：
Eisenmesser EZ

The Structural Dynamics, Complexity of Interactions, and Functions in Cancer of Multi-SAM Containing Proteins.

DOI：
10.3390/cancers15113019
发表时间：
2023-06-01
期刊：
Cancers
影响因子：
5.2
作者：
通讯作者：

Distance-independent Cross-correlated Relaxation and Isotropic Chemical Shift Modulation in Protein Dynamics Studies.

蛋白质动力学研究中距离无关的互相关弛豫和各向同性化学位移调制。

DOI：
10.1002/cphc.201800602
发表时间：
2019
期刊：
Chemphyschem : a European journal of chemical physics and physical chemistry
影响因子：
0
作者：
Vögeli,Beat;Vugmeyster,Liliya
通讯作者：
Vugmeyster,Liliya

Atomic resolution protein allostery from the multi-state structure of a PDZ domain.