Regulation of Cytoplasmic Dynein

细胞质动力蛋白的调节

• 批准号: 8630495
• 负责人: Andres Leschziner
• 金额: $ 48.66万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-10 至 2018-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8630495
• 关键词: ATP Hydrolysis; Address; Affect; Affinity; Beryllium; Binding; Biochemistry; Biological Assay; Biophysics; Cell physiology; Cells; Cellular biology; Complex; Cryoelectron Microscopy; Defect; Development; Disease; Dynein ATPase; Elements; Fluorescence Microscopy; Goals; Grant; Human; In Vitro; Kinesin; Laboratories; Lead; Mechanics; Mediating; Messenger RNA; Methods; Microtubules; Mitotic spindle; Modeling; Molecular; Molecular Motors; Motor; Motor Activity; Movement; Mutate; Mutation; Myosin ATPase; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Property; Proteins; Regulation; Research; Research Project Grants; Resolution; Role; Structure; Testing; Walking; base; cell motility; dimer; dynactin; in vivo; insight; macromolecule; monomer; mutant; polarized cell; protein complex; public health relevance; research study; single molecule; three dimensional structure

PROJECT SUMMARY The long-term goal of this research project is to understand how the microtubule (MT)-based motor cytoplasmic dynein ("dynein") is regulated. Dynein is the largest and most complex of all cytoskeletal motors; this >1 MDa dimeric complex contains numerous mechanical elements whose movements must be coordinated over strikingly long molecular distances to achieve processive motility along MTs. In addition to its enormous size, dynein is also the most versatile of the molecular motors; in sharp contrast to the 45 kinesins and 39 myosins present in humans, a single dynein gene product is responsible for transporting macromolecules within neurons, constructing the mitotic spindle, polarizing cells, and anchoring mRNAs during development. To give dynein the functional plasticity necessary for carrying out its many roles, several ubiquitous co-factors interact with dynein, including Lis1 and the dynactin complex. This project will apply our combined expertise in biochemistry, single-molecule biophysics and cryo-electron microscopy to address the structural and mechanistic bases of dynein's interaction with MTs and its regulation by Lis1 and dynactin. We recently showed that binding of dynein to MTs is accompanied by conformational changes in its MT- binding domain and that Lis1 acts as a "clutch" to uncouple MT binding and release from ATP hydrolysis, promoting a strongly MT-attached state. Dynactin, a 1.2 MDa complex, enhances dynein's processivity and is required for nearly all dynein functions in cells, but its mechanism of action is poorly understood. This grant will address major mechanistic questions about dynein and its regulation. What are the structural and mechanistic bases of MT binding (Aim 1), and of regulation by Lis1 (Aim 2) and dynactin (Aim 3)?

项目总结