Developmental control of organelle transport

细胞器运输的发育控制

• 批准号: 7525544
• 负责人: MICHAEL Andreas WELTE
• 金额: $ 27.97万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7525544
• 关键词: Affect; Alzheimer' s Disease; Binding; Biochemistry; Biological Models; Biological Process; Biology; Biophysics; Cardiovascular Diseases; Cells; Cellular biology; Destinations; Development; Disease; Drosophila genus; Dynein ATPase; Embryo; Embryonic Development; Ensure; Eukaryota; Eukaryotic Cell; Frequencies; Funding; Genetic; Genetic Techniques; Goals; Grant; Hand; Health; Human; Individual; Intracellular Transport; Kinesin; Light; Link; Lipids; Localized; Location; Malignant Neoplasms; Mammals; Maps; Measurement; Methods; Microtubules; Molecular; Molecular Biology; Motion; Motor; Obesity; Organelles; Phase; Play; Property; Protein Family; Proteins; Public Health; Range; Regulation; Research; Role; Signal Transduction; Signaling Protein; Specificity; Structure; System; Testing; Time; Transact; Transport Process; Travel; Work; design; dynactin; fly; fungus; in vivo; interdisciplinary approach; laser tweezer; member; mutant; nanoscale; novel; research study

DESCRIPTION (provided by applicant): The long-term goals of this proposal are to understand the regulatory mechanisms that allow organelle transport to be integrated with development. Motor-driven transport along microtubules plays many essential cellular roles, yet the mechanisms with which cells control its specificity, timing and destination remain a mystery. The proposal focuses on the rules of regulation for bidirectional transport, a kind of intracellular transport in which motor cooperation results in novel emergent phenomena not easily understood by studying the individual motors in isolation. A unique model system, lipid-droplet motion in Drosophila embryos, provides the opportunity to attack this long-standing problem in cell biology with an interdisciplinary approach that combines experimental techniques from genetics, biochemistry, molecular biology, and biochemistry. The proposal focuses on Phase II of droplet transport in which the Halo protein acts as a directionality determinant: in the absence of Halo, transport is minus-end directed; in the presence of Halo, it is plus-end directed. The questions asked are designed to unravel how the Halo signal is ultimately transmitted into changed activity of the plus-end motor kinesin-1 and the minus-end motor cytoplasmic dynein. Two complementary approaches are proposed, focusing on the one hand on the signal Halo and on the other hand on Sfo, a component of the motor machinery physically connected to the motors. 1) To understand how Halo acts as a signal, protein partners of Halo will be identified and a structure-function analysis of Halo will be performed. For selected Halo mutants, physical motion parameters will be quantified, using nanometer-scale tracking and stall-force measurements with optical tweezers. 2) Sfo has been proposed to be a "conductor", a molecule that fine-tunes how far cargoes travel in a single uninterrupted motion. Experiments are proposed to characterize the developmental expression and localization of Sfo, to determine if Sfo is required to transduce any aspects of the Halo signal, and to identify Sfo domains important in vivo. These experiments will in particular critically test whether Sfo indeed acts as a conductor. If successful, these studies will provide a framework for how a transacting signal is transduced by the transport machinery to affect properties of motors on a specific cargo. Because Halo is the founding member of a whole protein family in Drosophila, because Sfo is conserved from fungi to mammals and because both molecules are expressed throughout Drosophila development, these studies are expected to shed light on many different transport processes, both in flies and other eukaryotes. PUBLIC HEALTH RELEVANCE: The proposed work has broad significance for human health since aberrant motor function is linked to diseases ranging from Alzheimer's to cancer. These studies will also illuminate the basic biology of lipid droplets, organelles important in obesity and cardiovascular disease.

描述（由申请人提供）：该提案的长期目标是了解使Organelle运输与开发集成的监管机制。沿着微管的运动驱动运输扮演着许多必不可少的细胞角色，但是细胞控制其特异性，时机和目的地的机制仍然是一个谜。该提案的重点是双向转运的调节规则，这是一种细胞内运输，其中运动合作导致新型新兴现象不容易通过孤立研究单个电动机来理解。独特的模型系统，果蝇胚胎中的脂质动作运动为攻击细胞生物学中的这个长期存在的问题提供了跨学科方法，该方法结合了遗传学，生物化学，分子生物学和生物化学的实验技术。该提案的重点是液滴转运的II期，其中光环蛋白是方向性决定因素的：在没有光环的情况下，转运是负端的；在Halo面前，它是正式的。提出的问题旨在揭示如何最终将光晕信号传输到加号运动式驱动蛋白-1和减去末端运动细胞质动力蛋白的活性变化中。提出了两种互补方法，重点放在信号光环上，另一方面是SFO，SFO是电机机械的一个物理连接到电动机的组件。 1）要了解光环如何作为信号，将确定光环的蛋白质伴侣，并将对光环进行结构功能分析。对于选定的光晕突变体，使用纳米尺度跟踪和使用光学镊子进行摊位测量测量，将进行物理运动参数。 2）SFO已被认为是“指挥”，这是一种细微调的分子，可以通过单个不间断的运动进行货物的行驶程度。提出了实验来表征SFO的发育表达和定位，以确定是否需要SFO来转导光晕信号的任何方面，并确定体内重要的SFO域。这些实验将特别批判性地测试SFO是否确实充当导体。如果成功，这些研究将为运输机械转导的交易信号如何影响电动机在特定货物上的特性提供一个框架。因为Halo是果蝇中整个蛋白质家族的创始成员，因为SFO是从真菌到哺乳动物的保守的，并且由于两个分子在整个果蝇发育过程中都表达，因此这些研究有望阐明许多不同的运输过程，包括蝇和其他真核生物。 公共卫生相关性：拟议的工作对人类健康具有广泛的意义，因为异常运动功能与从阿尔茨海默氏症到癌症的疾病有关。这些研究还将阐明脂质液滴的基本生物学，对肥胖症和心血管疾病重要的细胞器。