Super-resolution studies of the entry mechanisms of influenza viruses
流感病毒侵入机制的超分辨率研究
基本信息
- 批准号:8040533
- 负责人:
- 金额:$ 33.78万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2004
- 资助国家:美国
- 起止时间:2004-09-22 至 2014-11-30
- 项目状态:已结题
- 来源:
- 关键词:ActinsAdaptor Signaling ProteinAreaBehaviorBindingBinding SitesBiochemicalBiological AssayBiologyBiomedical ResearchCell CommunicationCell physiologyCell surfaceCellsCellular StructuresCellular biologyClathrinClathrin-Coated VesiclesComplementDrug Delivery SystemsElectron MicroscopyEndocytosisEndosomesEventGlycolipidsGlycoproteinsGoalsHIVHealthHumanHuman Influenza A VirusHuman VirusImageImaging TechniquesImaging technologyIndividualInfectionInfluenzaInfluenza A Virus, H5N1 SubtypeInfluenza A virusIntegration Host FactorsInvestigationLaboratoriesLeadLengthLifeLipidsMediatingMicroscopyMolecularOpticsParasitesPathway interactionsPatientsPopulationProteinsRNA InterferenceResearchResolutionShapesSialic AcidsSpecificityStructureTFAP2A geneTechniquesTestingTimeVesicleVesicular stomatitis Indiana virusViralViral Envelope ProteinsViral GenomeVirionVirusVirus DiseasesVirus Replicationanti-influenzabasecoated pitepsin 1flufluorescence imaginggenome-wideimaging modalityimprovedinfluenza epidemicinfluenzavirusinsightmeetingsmolecular scalenanoscalenovelpandemic diseasepandemic influenzapathogenpolymerizationpreferenceprotein complexreceptorreconstructionresearch studytraffickingultra high resolutionvirology
项目摘要
DESCRIPTION (provided by applicant): Viral infections pose major threats to human health. As parasites that hijack endogenous cellular functions for replication, viruses are also excellent probes for studying cellular processes. Therefore investigations of viral infection mechanisms may not only lead to new therapies against viral diseases but also to a better understanding of fundamental cell biology and host cell-pathogen interactions. This application focuses on the influenza A virus. Our long-term goal is to dissect the influenza infection pathway and to understand the molecular mechanisms underlying individual steps along the infection pathway. In this project, we will focus on the viral entry step, which is a promising target for drug inhibition and for anti-flu therapies. Given the small sizes of the cellular endocytic structures and the virus, imaging with both ultra-high spatial resolution and molecular specificity is required to visualize the molecular details of viral entry. This requirement is difficult to meet with conventional imaging methods especially for living cells. We have recently developed a super-resolution fluorescence imaging technology, stochastic optical reconstruction microscopy (STORM), which provides near-molecular-scale resolution with high molecular specificity for imaging cellular structures. In this project, we will further improve the resolution of STORM and apply this novel technique to investigate the internalization mechanisms of influenza viruses for the first time. These high-resolution fluorescence imaging studies will be complemented by electron microscopy (EM), biochemical, cell biology, and virology assays. We will tackle the following four specific aims. In Aim 1, we will determine the spatio-temporal organization of the endocytic structures internalizing influenza virus, and compare the results with those obtained for other endocytic cargos, to elucidate the molecular mechanisms underlying virus internalization specifically and clathrin-mediated endocytosis in general. While influenza virus binds to cells through cell-surface sialic acids, various experiments suggest that influenza virus entry may require specific protein receptors in addition to sialic acids. In Aim 2, we will explore the molecular identity of the cellular receptor(s) required for influenza infection and investigate how such receptor(s) may guide viral entry. In Aims 3 and 4, we will determine the internalization mechanisms of two pathologically important influenza virus types. In Aim 3, we will investigate the internalization mechanism of the avian H5N1 influenza virus, which poses a pandemic threat, and compare the results to those on the human virus strains. In Aim 4, we will investigate the entry mechanism of filamentous influenza virus, which represents a prominent form of virus isolated from flu patients. We anticipate that results obtained from these studies will not only elucidate the entry mechanisms of influenza virus, but will also provide new insights into fundamental cellular processes, such as endocytosis, as well as host cell-pathogen interactions.
PUBLIC HEALTH RELEVANCE: In this project, we will use advanced imaging techniques with nanometer-scale resolution, together with biochemical, cell biology and virology assays, to investigate the infection mechanisms of influenza viruses, with particular emphasis on viral entry mechanisms. We anticipate that results from these studies will not only help elucidate the infection mechanisms of influenza viruses but will also provide new insights into fundamental cellular processes and into host cell-pathogen interactions. Furthermore, the super-resolution imaging approach developed here will have broad applications in many areas of biomedical research.
描述(由申请人提供):病毒感染对人类健康构成重大威胁。病毒作为劫持内源性细胞功能进行复制的寄生虫,也是研究细胞过程的优良探针。因此,对病毒感染机制的研究不仅可能导致针对病毒性疾病的新疗法,而且可能有助于更好地理解基础细胞生物学和宿主-病原体相互作用。此应用程序的重点是甲型流感病毒。我们的长期目标是剖析流感感染途径,并了解感染途径中单个步骤的分子机制。在这个项目中,我们将重点关注病毒进入步骤,这是药物抑制和抗流感治疗的一个有希望的目标。考虑到细胞内吞结构和病毒的小尺寸,需要超高空间分辨率和分子特异性的成像来可视化病毒进入的分子细节。这一要求是难以满足传统的成像方法,特别是对活细胞。我们最近开发了一种超分辨率荧光成像技术,随机光学重建显微镜(STORM),它为细胞结构成像提供了近分子尺度的分辨率和高分子特异性。在本项目中,我们将进一步提高STORM的分辨率,并首次将这种新技术应用于流感病毒内化机制的研究。这些高分辨率荧光成像研究将辅以电子显微镜(EM)、生化、细胞生物学和病毒学分析。具体抓好四个方面工作。在Aim 1中,我们将确定内化流感病毒的内吞结构的时空组织,并将结果与其他内吞货物的结果进行比较,以阐明病毒内化和网格蛋白介导的内吞作用的分子机制。虽然流感病毒通过细胞表面唾液酸与细胞结合,但各种实验表明,除了唾液酸外,流感病毒的进入可能还需要特定的蛋白质受体。在目标2中,我们将探索流感感染所需的细胞受体的分子特性,并研究这些受体如何引导病毒进入。在目标3和4中,我们将确定两种病理上重要的流感病毒类型的内化机制。在目标3中,我们将调查构成大流行威胁的H5N1禽流感病毒的内化机制,并将结果与人类病毒株的结果进行比较。在Aim 4中,我们将研究丝状流感病毒的进入机制,这是一种从流感患者中分离出来的突出的病毒形式。我们预计,从这些研究中获得的结果不仅将阐明流感病毒的进入机制,而且将为基本的细胞过程,如内吞作用以及宿主细胞-病原体相互作用提供新的见解。
项目成果
期刊论文数量(0)
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XIAOWEI ZHUANG其他文献
XIAOWEI ZHUANG的其他文献
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{{ truncateString('XIAOWEI ZHUANG', 18)}}的其他基金
Illuminating molecular mechanisms of cellular functions by single-molecule and super-resolution imaging
通过单分子和超分辨率成像阐明细胞功能的分子机制
- 批准号:
9275694 - 财政年份:2017
- 资助金额:
$ 33.78万 - 项目类别:
Illuminating molecular mechanisms of cellular functions by single-molecule and super-resolution imaging
通过单分子和超分辨率成像阐明细胞功能的分子机制
- 批准号:
9474629 - 财政年份:2017
- 资助金额:
$ 33.78万 - 项目类别:
Single-molecule studies of ATP-dependent chromatin remodeling
ATP依赖性染色质重塑的单分子研究
- 批准号:
8706191 - 财政年份:2013
- 资助金额:
$ 33.78万 - 项目类别:
Single-molecule studies of ATP-dependent chromatin remodeling
ATP依赖性染色质重塑的单分子研究
- 批准号:
9053499 - 财政年份:2013
- 资助金额:
$ 33.78万 - 项目类别:
Cellular Entry of Influenza by Single-particle Imaging
通过单粒子成像观察流感病毒的细胞进入
- 批准号:
7286279 - 财政年份:2004
- 资助金额:
$ 33.78万 - 项目类别:
Super-resolution studies of the entry mechanisms of influenza viruses
流感病毒侵入机制的超分辨率研究
- 批准号:
8204817 - 财政年份:2004
- 资助金额:
$ 33.78万 - 项目类别:
Super-resolution studies of the entry mechanisms of influenza viruses
流感病毒侵入机制的超分辨率研究
- 批准号:
8392262 - 财政年份:2004
- 资助金额:
$ 33.78万 - 项目类别:
Cellular Entry of Influenza by Single-particle Imaging
通过单粒子成像观察流感病毒的细胞进入
- 批准号:
6871654 - 财政年份:2004
- 资助金额:
$ 33.78万 - 项目类别:
Cellular Entry of Influenza by Single-particle Imaging
通过单粒子成像观察流感病毒的细胞进入
- 批准号:
7492222 - 财政年份:2004
- 资助金额:
$ 33.78万 - 项目类别:
Super-resolution studies of the entry mechanisms of influenza viruses
流感病毒侵入机制的超分辨率研究
- 批准号:
8588337 - 财政年份:2004
- 资助金额:
$ 33.78万 - 项目类别: