权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Mechanisms of microtubule-based transport

基于微管的运输机制

基本信息

批准号：
10661663
负责人：
SAMARA L RECK-PETERSON
金额：
$ 36.85万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
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：
2021-07-14 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10661663
关键词：
3-Dimensional Aspergillus nidulans Binding Biochemical Cells Chemicals Complex Cryo-electron tomography Cryoelectron Microscopy Cytoskeleton Defect Dynein ATPase Endosomes Eukaryotic Cell Family Genes Genetic Goals Human In Vitro Intracellular Transport Kinesin Link Melanophores Melanosomes Microtubules Molecular Molecular Machines Motor Movement Mutate Nature Neurodegenerative Disorders Neurodevelopmental Disorder Organelles Protein Engineering Proteins Proteomics RNA Editing Regulation Research Role Specific qualifier value Specificity Structure System Vertebrates Visualization Work Xenopus laevis cell motility disease-causing mutation dynactin fungal genetics genetic approach imaging genetics insight lissencephaly live cell imaging molecular imaging nervous system disorder ninein physical model programs reconstitution screening single molecule tool

项目摘要

PROJECT SUMMARY The contents of eukaryotic cells are highly dynamic, yet organized spatially and temporally. This is achieved primarily by the microtubule cytoskeleton and associated transport machinery, whose fundamental nature is highlighted by the many neurological diseases caused by mutations in them. The overarching goal of my research program is to understand how this system works at the molecular, cellular, and organismal scales. My team is highly interdisciplinary and we use in vitro biochemical reconstitution, protein engineering, single-molecule imaging, proteomics, live-cell imaging, and fungal genetics to achieve our goals. Through collaborative projects we use cryo-electron microscopy (cryo-EM) and cryo-electron tomography (cryo-ET) to incorporate a structure-guided approach to understanding intracellular transport, and we develop testable quantitative physical models of transport. We have made major contributions to determining how the dynein motor works and is regulated, to developing tools and screening strategies to study bi-directional movement of cargos on microtubules, and to understanding the regulation of intracellular transport in cells. Fundamental questions that we will address here include: (1) How does the dynein motor work? Our earlier work revealed how Lis1, a protein mutated in the neurodevelopmental disease lissencephaly, interacts with dynein and regulates its mechanochemical cycle. Here, we will focus on determining the mechanistic underpinnings for how Lis1 promotes the formation of activated dynein/dynactin complexes. We will also explore a new direction—the role of RNA editing—as a previously undescribed mechanism to regulate dynein and kinesin motors. Microtubule-based motors move dozens if not hundreds of cargos. (2) How is cargo-specificity achieved? Our past work used two complementary discovery-based approaches—genetics and proteomics—to identify molecules responsible for specifying dynein’s many functions. One mechanism revealed by our past work is organelle hitchhiking, where cargos link to motors indirectly, by attaching themselves to other cargos that are directly bound to the motors. A second strategy for achieving cargo specificity is the expansion of dynein activating adaptor genes in vertebrates. However, the molecular connections between most activating adaptors and dynein’s cargo are unknown. Here, we will determine the mechanisms underlying hitchhiking and the linkages between the Hook and Ninein families of activating adaptors and their cargos. As an additional approach to understand how dynein and kinesin link to their cargos, we will visualize these connections in cells in three dimensions using cryo-electron tomography of endosomes in Aspergillus nidulans and melanosomes in Xenopus laevis melanophores, two systems where we can use exquisite genetics or chemical tools to control microtubule- based motility.

项目概要真核细胞的内容是高度动态的，但在空间和时间上是有组织的。这样就实现了主要由微管细胞骨架和相关的运输机制组成，其基本性质是突出的是由它们的突变引起的许多神经系统疾病。我的总体目标研究计划是为了了解该系统如何在分子、细胞和有机体方面发挥作用秤。我的团队是高度跨学科的，我们使用体外生化重建、蛋白质工程、单分子成像、蛋白质组学、活细胞成像和真菌遗传学来实现我们的目标。通过我们使用冷冻电子显微镜 (cryo-EM) 和冷冻电子断层扫描 (cryo-ET) 的合作项目结合结构引导的方法来理解细胞内运输，我们开发了可测试的运输的定量物理模型。我们为确定动力蛋白如何发挥作用做出了重大贡献运动工作并受到监管，开发工具和筛选策略来研究双向运动微管上的货物，并了解细胞内运输的调节。基本的我们在这里要解决的问题包括：（1）动力蛋白电机如何工作？我们早期的工作揭示了 Lis1（一种在神经发育疾病无脑畸形中发生突变的蛋白质）如何与动力蛋白相互作用调节其机械化学循环。在这里，我们将重点确定如何实现的机制基础 Lis1 促进活化动力蛋白/动力蛋白复合物的形成。我们还将探索一个新的方向—— RNA 编辑的作用——作为一种先前未描述的调节动力蛋白和驱动蛋白马达的机制。基于微管的马达可以移动数十甚至数百件货物。 (2) 货物专用性是如何实现的？我们的过去的工作使用了两种互补的基于发现的方法——遗传学和蛋白质组学——来识别负责指定动力蛋白许多功能的分子。我们过去的工作揭示的一种机制是细胞器搭便车，货物通过将自身附着到其他货物上来间接连接到发动机直接绑定到电机。实现货物特异性的第二个策略是扩展动力蛋白激活脊椎动物中的接头基因。然而，大多数激活接头之间的分子连接动力蛋白的货物未知。在这里，我们将确定搭便车的潜在机制和 Hook 和 Ninein 系列激活适配器与其货物之间的联系。作为一种额外的方法为了了解动力蛋白和驱动蛋白如何与其货物连接，我们将在三个方面可视化细胞中的这些连接使用冷冻电子断层扫描分析构巢曲霉内体和非洲爪蟾黑素体的尺寸非洲兔黑素细胞，我们可以使用精致的遗传学或化学工具来控制微管的两个系统- 基于运动性。

项目成果

期刊论文数量（9）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Structures of human dynein in complex with the lissencephaly 1 protein, LIS1.

DOI：
10.7554/elife.84302
发表时间：
2023-01-24
期刊：
ELIFE
影响因子：
7.7
作者：
Reimer, Janice M.;DeSantis, Morgan E.;Leschziner, Andres E.;Reck-Peterson, Samara L.
通讯作者：
Reck-Peterson, Samara L.

Lis1 relieves cytoplasmic dynein-1 autoinhibition by acting as a molecular wedge.

DOI：
10.1038/s41594-023-01069-6
发表时间：
2023-09
期刊：
NATURE STRUCTURAL & MOLECULAR BIOLOGY
影响因子：
16.8
作者：
Karasmanis, Eva P.;Reimer, Janice M.;Kendrick, Agnieszka A.;Nguyen, Kendrick H. V.;Rodriguez, Jennifer A.;Truong, Joey B.;Lahiri, Indrajit;Reck-Peterson, Samara L.;Leschziner, Andres E.
通讯作者：
Leschziner, Andres E.

Woronin body hitchhiking on early endosomes is dispensable for septal localization in Aspergillus nidulans.

DOI：
10.1091/mbc.e23-01-0025
发表时间：
2023-06-01
期刊：
MOLECULAR BIOLOGY OF THE CELL
影响因子：
3.3
作者：
Songster, Livia D.;Bhuyan, Devahuti;Christensen, Jenna R.;Reck-Peterson, Samara L.
通讯作者：
Reck-Peterson, Samara L.

Woronin bodies move dynamically and bidirectionally by hitchhiking on early endosomes in Aspergillus nidulans.

Woronin 体通过搭便车在构巢曲霉的早期内体上进行动态和双向移动。

DOI：
10.1101/2023.01.20.524968
发表时间：
2023
期刊：
bioRxiv : the preprint server for biology
影响因子：
0
作者：
Songster,LiviaD;Bhuyan,Devahuti;Christensen,JennaR;Reck-Peterson,SamaraL
通讯作者：
Reck-Peterson,SamaraL

PxdA interacts with the DipA phosphatase to regulate peroxisome hitchhiking on early endosomes.

DOI：
10.1091/mbc.e20-08-0559
发表时间：
2021-03-15
期刊：
Molecular biology of the cell
影响因子：
3.3
作者：
Salogiannis J;Christensen JR;Songster LD;Aguilar-Maldonado A;Shukla N;Reck-Peterson SL
通讯作者：
Reck-Peterson SL