Recruitment of BST-2/tetherin to HIV-1 assembly sites

将 BST-2/tetherin 招募到 HIV-1 装配位点

• 批准号: 8210153
• 负责人: Akira Ono
• 金额: $ 18.87万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8210153
• 关键词: Acquired Immunodeficiency Syndrome; Antibodies; Antiviral Agents; Arenavirus; Biology; C-terminal; Carrier Proteins; Cell membrane; Cell physiology; Cell surface; Cells; Cellular Structures; Cholesterol; Complex; Cytoplasmic Tail; Data; Data Reporting; Defect; Detergents; Event; Family; Filovirus; Fluorescence; Fluorescence Microscopy; Gagging; Genetic; Goals; HIV; HIV-1; Herpesviridae; Integral Membrane Protein; Knowledge; Label; Life; Link; Lipid Bilayers; Lipids; Mediating; Membrane; Membrane Microdomains; Microscopy; Modeling; N-terminal; Neck; Paramyxovirus; Planning Techniques; Play; Proteins; Recruitment Activity; Research; Resistance; Resolution; Retroviridae; Rhabdoviridae; Role; Satellite Viruses; Scanning Electron Microscopy; Sequence Homology; Site; Sorting - Cell Movement; Structural Protein; Structure; Techniques; Testing; Transmembrane Domain; Vial device; Viral; Virion; Virus; Virus Assembly; cell fixing; design; extracellular; interest; mutant; novel; particle; research study; single molecule

DESCRIPTION (provided by applicant): Recent studies showed that a cellular protein known as HM1.24, CD317, BST-2, or tetherin (hereafter referred to as BST-2/tetherin) inhibits HIV-1 particle release by tethering newly assembled virus particles to the plasma membrane (PM) of virus-producing cells. However, the mechanism by which BST-2/tetherin is incorporated into assembling virus particles remains to be determined. Notably, BST-2/tetherin can inhibit release of a wide variety of enveloped viruses including retroviruses, filoviruses, herpesviruses, rhabdoviruses, paramyxoviruses, and arenaviruses. Therefore, determining the mechanism that recruits BST-2/tetherin to HIV-1 assembly sites will not only advance our understanding of the biology of HIV that causes AIDS but also will help us develop antiviral strategies targeting a broad range of enveloped viruses. Notably, an artificial protein that shares key structural features with BST-2/tetherin with no sequence homology was capable of blocking HIV-1 release. Therefore, association of BST-2/tetherin to nascent virus particles is likely dependent on a common cellular structure rather than specific viral or cellular proteins. The PM is now thought to consist of multiple microdomains rather than a uniform lipid bilayer with proteins embedded in a random manner. These microdomains contain specific sets of lipids and proteins and may have various lifetimes, sizes, and dynamics. Among them, lipid rafts are well known to associate with HIV-1 assembly. Notably, BST-2/tetherin is modified at its C terminus with a GPI anchor, which is known to target proteins to lipid rafts. Together with the notion that many enveloped viruses associate with lipid rafts or other microdomains, it is conceivable that association of BST-2/tetherin with lipid rafts may promote incorporation of this protein to assembling particles. However, it is also possible that other membrane-associated structures common to BST-2/tetherin-susceptible viruses, such as membrane curvature, membranous neck/stalk, or ESCRT (endosomal sorting complex required for transport) proteins, play a role in BST-2/tetherin recruitment. Toward better understanding of the antiviral activity of BST-2/tetherin, in this proposal, we plan to test the following hypothesis: BST-2/tetherin associates with specific PM structures that are recruited to virus assembly sites. To test this hypothesis, in Aim 1, the order of events with regard to incorporation of rafts, ESCRTs, and BST-2/tetherin during virus assembly will be determined. Using combination of genetic approaches with advanced microscopy techniques, we plan to identify the assembly step at which BST-2/tetherin are recruited to assembly sites. Experiments in Aim 2 are designed to determine whether lipid rafts and ESCRTs are essential for BST- 2/tetherin function and its recruitment to virus assembly sites. Altogether, information gained in experiments proposed in this application will likely help us elucidate the mechanism that initiates BST-2/tetherin-mediated restriction of HIV-1 release. PUBLIC HEALTH RELEVANCE: BST-2/tetherin is a protein that inhibits release of HIV-1 and other enveloped viruses from infected cells by physically linking virus particles to the cell surface membrane. The goal of the proposed research is to elucidate the role played by the cell surface membrane structures common to many enveloped viruses during incorporation of BST-2/tetherin into forming virus particles. Knowledge obtained from proposed studies will help us develop novel antiviral strategies that may suppress spread of not only HIV-1 that causes AIDS but also a wide variety of enveloped viruses.

描述（由申请人提供）：最近的研究表明，一种称为HM1.24，CD317，BST-2或Tetherin（以下称为BST-2/Tetherin）的细胞蛋白通过将新组装的病毒颗粒固定在质膜（PM）的病毒膜（PM）中，抑制HIV-1颗粒释放。但是，将BST-2/Tetherin纳入组装病毒颗粒的机制仍有待确定。值得注意的是，BST-2/Tetherin可以抑制各种包膜病毒的释放，包括逆转录病毒，丝状病毒，疱疹病毒，黄脂病毒，帕拉马病毒和体育症病毒。因此，确定将BST-2/Tetherin招募到HIV-1组装部位的机制不仅会提高我们对引起艾滋病的HIV生物学的理解，而且还将帮助我们制定针对广泛包膜病毒的抗病毒毒剂策略。值得注意的是，具有与BST-2/Tetherin共享关键结构特征的人工蛋白质无序性同源性能够阻止HIV-1释放。因此，BST-2/Tetherin与新生病毒颗粒的关联可能取决于常见的细胞结构，而不是特定的病毒或细胞蛋白。现在，PM被认为由多个微区域而不是均匀的蛋白质组成。这些微区域包含特定的脂质和蛋白质集，并且可能具有各种寿命，大小和动力学。其中，脂质筏众所周知，可以与HIV-1组装相关。值得注意的是，BST-2/Tetherin用GPI锚在其C末端进行了修饰，GPI锚定为蛋白质靶向脂质筏。加上许多包裹病毒与脂质筏或其他微域相关的概念，可以想象，BST-2/Tetherin与脂质筏的缔合可能促进该蛋白质与组装颗粒的结合。然而，也有可能与BST-2/Tetherin敏感病毒共有的其他相关结构，例如膜曲率，膜颈/茎/茎或ESCRT或ESCRT（运输所需的内体分类复合物）蛋白，在BST-2/TETHETHERIN招募中起作用。为了更好地理解BST-2/Tetherin的抗病毒活性，在此提案中，我们计划检验以下假设：BST-2/Tetherin与特定的PM结构相关，这些结构被募集到病毒组装位点。为了检验该假设，在AIM 1中，将确定病毒组装过程中有关筏，ESCRT和BST-2/Tetherin的事件的顺序。使用遗传方法与高级显微镜技术的组合，我们计划确定将BST-2/Tetherin招募到组装位点的组装步骤。 AIM 2中的实验旨在确定脂质筏和ESCRT是否对于BST-2/Tetherin功能及其募集到病毒组装位点至关重要。总之，在本应用程序中提出的实验中获得的信息可能会帮助我们阐明启动BST-2/Tetherin介导的HIV-1释放限制的机制。 公共卫生相关性：BST-2/Tetherin是一种蛋白质，可通过将病毒颗粒与细胞表面膜物理联系到感染细胞中释放HIV-1和其他包膜病毒。拟议的研究的目的是阐明在将BST-2/Tetherin掺入形成病毒颗粒的过程中，许多包膜病毒常见的细胞表面膜结构所起的作用。从拟议的研究中获得的知识将有助于我们制定新颖的抗病毒药策略，这些抗病毒病毒策略不仅可以抑制引起艾滋病的HIV-1的传播，还可以抑制各种包裹的病毒。