Polyomavirus transport: Vesicles, motor proteins, and endocytosis.

多瘤病毒运输：囊泡、运动蛋白和内吞作用。

• 批准号: 7212592
• 负责人: CHRISTINE K PAYNE
• 金额: $ 16.1万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-06-15 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7212592
• 关键词: Actins; Affect; Avidin; Behavior; Biological Assay; Biotin; Caveolins; Cells; Cessation of life; Cholera Toxin; Clathrin; Color; Coma; Complex; Condition; Cytoskeletal Proteins; Cytoskeleton; DNA Sequence Rearrangement; Dementia; Dependence; Development; Disruption; Drug Delivery Systems; Dynein ATPase; Endocytosis; Endosomes; Fluorescence Microscopy; Fluorescent Probes; Genetic; Goals; Green Fluorescent Proteins; Health; Human; Image; Imaging Techniques; Imaging technology; Immunocompromised Host; Individual; Infection; Intracellular Transport; Investigation; JC Virus; Kinesin; Label; Lasers; Life; Link; Lipids; Location; Low-Density Lipoproteins; Measurement; Measures; Mediating; Methods; Microscopy; Microtubules; Motion; Motor; Mus; Myelin; Myosin ATPase; Noise; Numbers; Optical Methods; Pathway interactions; Peptides; Personal Satisfaction; Persons; Pharmaceutical Preparations; Polymers; Polyomavirus; Population; Principal Investigator; Progressive Multifocal Leukoencephalopathy; Proteins; Purpose; Quantum Dots; Range; Research; Research Personnel; Resolution; Role; Signal Transduction; Simian virus 40; Sorting - Cell Movement; Source; Speed; System; Techniques; Testing; Time; Transport Vesicles; Vesicle; Viral; Virus; Virus Diseases; Visible Radiation; Work; base; caveolin 1; cellular imaging; charge coupled device camera; fluorescence imaging; fluorescence microscope; interest; late endosome; mouse polyomavirus; mutant; nanometer; particle; programs; receptor; receptor mediated endocytosis; research study; size

DESCRIPTION (provided by applicant): An understanding of the human polyomavirus, JC virus, is of great importance due to its active infection of immunocompromised persons. Active infection results in progressive multifocal leukoencephalopathy (PML). PML causes the destruction of myelin resulting in dementia, coma, and death. The intracellular pathway of JC virus has not been determined, providing a barrier to the treatment of active infection. The goal of this investigation is to determine how JC and other polyomaviruses are transported in cells. This includes determining the vesicles used for transport and the motor proteins that drive the motion. I propose that the transport of specific populations of vesicles is independent of the cargo contained. This hypothesis will be tested in two steps: 1. The human, simian, and murine polyomaviruses will be imaged during intracellular transport to determine which vesicles and motor proteins are used. Two- and three-color fluorescence imaging will be used to determine which vesicles the virus uses for transport. Characterization of the motor proteins involved in transport will be based on the individual steps of the virus on the cytoskeleton. These experiments will be the first to make use of a recently developed technique, Fluorescence Imaging with One Nanometer Accuracy (FIONA), to probe viral transport on the level of individual motor protein steps. 2. The experimental techniques developed will then be extended to non-viral cargo to test the general relationship between transport dynamics and vesicle population. To this end the polyomaviruses will serve as probes of the three major endocytic pathways. Non-viral cargo will be loaded into specific vesicles and imaged during intracellular transport. The dynamics observed will be compared to those of the polyomaviruses. Overall, this research is expected to contribute significantly to our understanding of three interrelated fields; viral infection, imaging technology, and intracellular transport.

描述(由申请人提供)：了解人类多瘤病毒JC病毒非常重要，因为它会主动感染免疫功能低下的人。活动性感染导致进行性多灶性白质脑病(PML)。PML导致髓鞘破坏，导致痴呆、昏迷和死亡。JC病毒的细胞内途径尚未确定，这为活动性感染的治疗提供了障碍。这项调查的目标是确定JC和其他多瘤病毒如何在细胞中传播。这包括确定用于运输的囊泡和驱动运动的马达蛋白。我认为，特定群体的囊泡的运输与所含货物无关。这一假设将分两个步骤进行检验：1.在细胞内转运过程中，将对人类、猿类和小鼠多瘤病毒进行成像，以确定使用了哪些囊泡和马达蛋白。将使用双色和三色荧光成像来确定病毒使用哪些小泡进行运输。对参与运输的马达蛋白的表征将基于病毒在细胞骨架上的个别步骤。这些实验将首次利用最近开发的一种技术--一纳米精度荧光成像(Fiona)--在单个运动蛋白质步骤的水平上探测病毒的运输。2.开发的实验技术将扩展到非病毒货物，以测试运输动力学和囊泡数量之间的一般关系。为此，多瘤病毒将作为三条主要内吞途径的探针。非病毒货物将被装载到特定的囊泡中，并在细胞内运输过程中进行成像。观察到的动态将与多瘤病毒的动态进行比较。总体而言，这项研究有望对我们理解三个相互关联的领域做出重大贡献：病毒感染、成像技术和细胞内运输。