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

• 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
• 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
• 来源: https://reporter.nih.gov/project-details/10680430
• 关键词: ATP Hydrolysis; Actins; Alzheimer' s Disease; Architecture; Behavior; Binding; Biochemistry; Biological; Biological Assay; Cells; Chemicals; Complex; Cytoplasm; Cytoskeleton; Defect; Destinations; Disease; Dynein ATPase; Environment; Eukaryota; Family; Filament; Goals; Health; Homeostasis; Human; Huntington Disease; Impairment; In Vitro; Individual; Intracellular Transport; Kinesin; Lead; Link; Microtubule-Associated Proteins; Microtubules; Molecular; Molecular Motors; Motion; Motor; Motor Activity; Motor output; Neurodegenerative Disorders; Pathologic; Pick Disease of the Brain; Process; Property; Protein Dynamics; Regulation; Research; System; Testing; Translating; Work; cell motility; human disease; in vivo; insight; novel; reconstitution; recruit; scaffold; single molecule; spatiotemporal; tau Proteins; tool

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：细胞内转运和细胞骨架组装中分子马达活性的协调 架构 P.I. - 理查德·J·麦肯尼 研究综述 细胞内转运是真核生物细胞内环境稳定所必需的。这一过程是在 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 和微管细胞骨架网络。几十年的研究揭示了结构和分子 这些细节解释了这些马达中有多少沿着它们的灯丝轨道沿着移动。在细胞环境中， 这些马达中的大多数与复杂的调节机制相一致， 相应的货物，调节其运动特性，并决定时空活动。如何 单个电机输出是由这种机械控制的，目前尚不清楚，也难以剖析。 细胞的复杂环境。此外，许多货物是通过协调行动， 同时绑定，相反极性的电机，在这个过程中称为双向运输。如何 单个发动机被招募到货物上，激活，并与其他类别的发动机集成在一起 对该领域提出了巨大的挑战。重要的是，这一过程中的缺陷导致各种各样的 这些问题直接关系到人类的健康。 微管网络的组织、动力学和沿着微管的运动都是 受到非酶促微管相关蛋白（MAP）的影响，这些蛋白动态地结合到 微管MAPs的具体功能、分子特性和动力学仍有待进一步研究 探索不足tau家族的MAP对人类健康至关重要，因为tau是很好的- 其特征在于在许多人类神经变性疾病中形成不溶性内含物/聚集体 如阿尔茨海默病和皮克病。尽管他们四十多年前就被确认了， tau家族MAPs的特异性分子功能和分子特性仍不清楚。这 申请旨在开发新的测定和工具来研究微管马达的复杂性， 调节、货物的双向运输以及由马达和MAP驱动的细胞骨架功能。 我们将生物化学和单分子分析联合收割机用于体外重组 测试分子功能，并将我们的发现转化为体内系统， 这些重组产生的成果将开辟富有成效的长期研究途径。我们提出 目的：1）重建和研究细胞质动力蛋白的募集、调节和运动， 与天然细胞货物支架分子结合的驱动蛋白马达，和2）重建和研究 tau家族MAPs的进化功能、动力学和病理行为。这些广泛的目标 建立和扩大我们的专业知识和以前的工作，在解剖的监管机制， 细胞质动力蛋白马达，旨在提供强大的新工具， 复杂运动功能我们的工作将阐明基本的分子和细胞生物学原理， 驱动细胞内稳态，并提供对病理机制的洞察， 分子马达故障

项目成果

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.)
• 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.

神经元膜交通的极性需要在微管相关的SEPTIN进入树突期间对运动蛋白运动货物进行排序。

• 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
• 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。

• DOI: 10.1038/s41556-020-0500-5
• 发表时间: 2020
• 期刊: Nature cell biology
• 影响因子: 21.3
• 作者: McKenney,RichardJ