Coordination of molecular motor activity in intracellular transport and assembly of cytoskeletal archit

细胞内运输和细胞骨架结构组装中分子运动活动的协调

基本信息

批准号：
10680430
负责人：
Richard James McKenney
金额：
$ 42.92万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-30 至 2027-08-31
项目状态：
未结题

项目摘要

Title: Coordination of molecular motor activity in intracellular transport and assembly of cytoskeletal architecture. P.I. – Richard J. McKenney Research Summary Intracellular transport is essential for cellular homeostasis in eukaryotes. This process is carried out by molecular motors that convert the chemical energy from ATP hydrolysis into motion along the actin and microtubule cytoskeletal networks. Decades of research has uncovered structural and molecular details that explain how many of these motors move along their filament tracks. In the cellular milieu, most of these motors act in concert with complex regulatory machinery that links them to their respective cargos, modulates their motile properties, and dictates spatiotemporal activity. How individual motor output is controlled by this machinery is currently not clear and difficult to dissect in the complex environment of the cell. In addition, many cargos are moved by the concerted action of simultaneously bound, opposite polarity motors, in a process called bidirectional transport. How individual motors are recruited to cargo, activated, and integrated with other classes of motors presents a large open challenge to the field. Importantly, defects in this process lead to a wide variety of human diseases and these questions are thus directly related to human health. Microtubule network organization, dynamics, and motor activity along microtubules are all impinged upon by non-enzymatic microtubule-associated proteins (MAPs) that dynamically bind to microtubules. The specific functions, molecular properties, and dynamics of MAPs remains underexplored. The tau family of MAPs are critically important for human health, as tau is well- characterized to form insoluble inclusions/aggregates in a host of human neurodegenerative diseases such as Alzheimer’s disease and Pick’s disease. Despite their identification over four decades ago, the specific molecular functions and molecular properties of tau family MAPs remains unclear. This application seeks to develop novel assays and tools to study the complexity of microtubule motor regulation, bidirectional transport of cargos, and cytoskeletal functions driven by motors and MAPs. Our approach to combine biochemistry and single-molecule analysis towards in vitro reconstitutions that test molecular function, and translate our findings into in vivo systems that test hypotheses generated by these reconstitutions, will open up fruitful long-term avenues of research. We propose to: 1) Reconstitute and study the recruitment, regulation, and motility of cytoplasmic dynein and kinesin motors bound to native cellular cargo scaffolding molecules, and 2) Reconstitute and study evolutionary functions, dynamics, and pathological behaviors of tau family MAPs. These broad goals build and expand upon our expertise and previous work in dissecting the regulatory mechanisms of the cytoplasmic dynein motor, and aim to provide powerful new tools useful towards dissecting complex motor function. Our work will illuminate basic molecular and cell biological principles that drive cellular homeostasis and provide insight into the pathological mechanisms that arise from molecular motor malfunction.

标题：细胞内运输和细胞骨架组装中分子运动活动的协调建筑学。 P.I.——理查德·J·麦肯尼研究总结细胞内运输对于真核生物的细胞稳态至关重要。通过分子马达将 ATP 水解产生的化学能转化为沿着肌动蛋白的运动数十年的研究揭示了结构和分子网络。解释了在细胞环境中有多少电机沿着细丝轨道移动的细节。这些电机中的大多数与复杂的监管机制协同作用，将它们与它们的各自的货物，调节它们的运动特性，并决定时空活动。单个电机的输出是由该机器产生的，目前尚不清楚且难以剖析此外，许多货物是通过细胞的协同作用而移动的。同时绑定相反极性的电机，这一过程称为双向传输。各个电机被招募到货物、激活并与其他类别的电机集成重要的是，这个过程中的缺陷导致了各种各样的问题。因此，这些问题与人类健康直接相关。微管网络组织、动力学和沿着微管的运动活动都是受到动态结合的非酶微管相关蛋白 (MAP) 的影响微管的特定功能、分子特性和动力学仍然存在。 MAP 的 tau 蛋白家族对人类健康至关重要，因为 tau 蛋白已被广泛研究。其特征是在许多人类神经退行性疾病中形成不溶性内含物/聚集体例如阿尔茨海默病和皮克病，尽管它们在四十多年前就被识别出来了， tau家族MAPs的具体分子功能和分子特性仍不清楚。该申请旨在开发新的分析方法和工具来研究微管运动的复杂性调节、货物双向运输以及由电机和 MAP 驱动的细胞骨架功能。我们将生物化学和单分子分析结合起来进行体外重组的方法测试分子功能，并将我们的发现转化为测试假设的体内系统我们建议，这些重组所产生的成果将开辟富有成效的长期研究途径。 1) 重建和研究细胞质动力蛋白的募集、调节和运动驱动蛋白马达与天然细胞货物支架分子结合，2) 重建和研究 tau 家族 MAP 的进化功能、动力学和病理行为。建立并扩展我们的专业知识和之前在剖析监管机制方面的工作细胞质动力蛋白马达，旨在提供有助于解剖的强大新工具我们的工作将阐明基本的分子和细胞生物学原理。驱动细胞稳态并深入了解由分子马达故障。

项目成果

期刊论文数量（23）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Microtubule lattice spacing governs cohesive envelope formation of tau family proteins.

DOI：
10.1038/s41589-022-01096-2
发表时间：
2022-11
期刊：
NATURE CHEMICAL BIOLOGY
影响因子：
14.8
作者：
Siahaan, Valerie;Tan, Ruensern;Humhalova, Tereza;Libusova, Lenka;Lacey, Samuel E.;Tan, Tracy;Dacy, Mariah;Ori-McKenney, Kassandra M.;McKenney, Richard J.;Braun, Marcus;Lansky, Zdenek
通讯作者：
Lansky, Zdenek

Tau oligomerization on microtubules in health and disease.

健康和疾病中微管的 Tau 寡聚化。

DOI：
10.1002/cm.21785
发表时间：
2024
期刊：
Cytoskeleton (Hoboken, N.J.)
影响因子：
0
作者：
Ori-McKenney,KassandraM;McKenney,RichardJ
通讯作者：
McKenney,RichardJ

Polarity of Neuronal Membrane Traffic Requires Sorting of Kinesin Motor Cargo during Entry into Dendrites by a Microtubule-Associated Septin.

DOI：
10.1016/j.devcel.2018.06.013
发表时间：
2018-07-16
期刊：
Developmental cell
影响因子：
11.8
作者：
Karasmanis EP;Phan CT;Angelis D;Kesisova IA;Hoogenraad CC;McKenney RJ;Spiliotis ET
通讯作者：
Spiliotis ET

Evidence for anaphase pulling forces during C. elegans meiosis.

DOI：
10.1083/jcb.202005179
发表时间：
2020-12-07
期刊：
The Journal of cell biology
影响因子：
0
作者：
Danlasky BM;Panzica MT;McNally KP;Vargas E;Bailey C;Li W;Gong T;Fishman ES;Jiang X;McNally FJ
通讯作者：
McNally FJ

LIS1 cracks open dynein.

LIS1 裂解动力蛋白。