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The role of virus structure and host cell membrane trafficking as well as signalling factors in Human Papillomavirus Type 16 endocytosis

病毒结构和宿主细胞膜运输以及信号因子在人乳头瘤病毒16型内吞作用中的作用

基本信息

批准号：
83146105
负责人：
Professor Dr. Mario Schelhaas
金额：
--
依托单位：
Assoziiertes Institut für Zelluläre Virologie
依托单位国家：
德国
项目类别：
Independent Junior Research Groups
财政年份：
2008
资助国家：
德国
起止时间：
2007-12-31 至 2014-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/83146105?language=en
关键词：
role virus structure host cell

项目摘要

Several human papillomaviruses (HPV) are the central cause for cervical cancer. The major HPV subtype found in cervix carcinomas is HPV-16. Little is known about the entry of HPVs into host cells except that it occurs via endocytosis. Our results indicate that HPV-16 enters host cells by a clathrin- and caveolin-independent mechanism that features some but not all characteristics of macropinocytosis, and that likely represents a novel endocytic route into cells. We propose to further investigate the cellular entry of HPV-16 by endocytosis. This project has the following aims: - To identify most of the host cell factors involved in HPV-16 endocytosis- To elucidate the role of host cell factors and signalling in the dynamics of HPV-16 endocytosis- To understand the molecular role of sulfated carbohydrates in binding of HPV-16- To identify structural changes in HPV-16 virions that serve as functional ‚cues’ for endocytic entry and trafficking as well as for uncoating of the virus.The main methods to be applied include various techniques for live cell microscopy as well as cell biological, morhological, genetic, and biochemical techniques. To categorise the requirements for HPV entry a genome-wide siRNA-mediated gene silencing screen will be performed in an automated high through-put assay. To gain temporal and spatial information on the events in HPV endocytosis, several advanced light microscopic techniques (wide field, confocal, and total internal reflection microscopy) will be used to follow single HPVs and their association with specific markers for endocytic organelles in living cells. To determine the carbohydrates that serve most specifically for binding of HPV-16 to host cells we will screen a carbohydrate microarrays. To elucidate structural changes in virus particles during endocytosis we will use isolation, biochemical, and ultrastructural analysis of virus particles from infected cells. The results are expected to contribute both to the understanding of HPV entry into host cells, and to the analysis of endocytic traffic in mammalian cells.

几种人乳头瘤病毒（HPV）是宫颈癌的主要原因。在宫颈癌中发现的主要HPV亚型是HPV-16。除了通过内吞作用外，人们对hpv进入宿主细胞知之甚少。我们的研究结果表明，HPV-16通过一种不依赖网格蛋白和小窝蛋白的机制进入宿主细胞，这种机制具有巨噬细胞症的一些特征，但不是全部特征，这可能代表了一种新的细胞内吞途径。我们建议进一步研究HPV-16通过内吞作用进入细胞。该项目有以下目标：-确定参与HPV-16内吞作用的大多数宿主细胞因子-阐明宿主细胞因子和信号在HPV-16内吞作用动力学中的作用-了解硫酸碳水化合物在HPV-16结合中的分子作用-确定HPV-16病毒粒子的结构变化，这些结构变化可作为内吞进入和运输以及病毒脱壳的功能线索。应用的主要方法包括各种活细胞显微镜技术以及细胞生物学、形态学、遗传学和生化技术。为了对HPV进入的要求进行分类，将在自动化的高通量分析中进行全基因组sirna介导的基因沉默筛选。为了获得HPV内吞事件的时间和空间信息，将使用几种先进的光学显微镜技术（宽视野，共聚焦和全内反射显微镜）来跟踪单个HPV及其与活细胞内吞细胞器特定标记物的关联。为了确定对HPV-16与宿主细胞结合最特异的碳水化合物，我们将筛选碳水化合物微阵列。为了阐明病毒颗粒在内吞过程中的结构变化，我们将对感染细胞中的病毒颗粒进行分离、生化和超微结构分析。该结果有望有助于理解HPV进入宿主细胞，并分析哺乳动物细胞的内吞运输。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Heparin increases the infectivity of Human Papillomavirus type 16 independent of cell surface proteoglycans and induces L1 epitope exposure.

肝素增加了独立于细胞表面蛋白聚糖的16型人乳头瘤病毒的感染性，并诱导L1表位暴露。

DOI：
10.1111/cmi.12150
发表时间：
2013-11
期刊：
Cellular microbiology
影响因子：
3.4
作者：
Cerqueira C;Liu Y;Kühling L;Chai W;Hafezi W;van Kuppevelt TH;Kühn JE;Feizi T;Schelhaas M
通讯作者：
Schelhaas M

Concepts of papillomavirus entry into host cells.