GENETICS OF PRIMATE 'D' TYPE RETROVIRUSES

灵长类“D”型逆转录病毒的遗传学

• 批准号: 8357425
• 负责人: Eric Hunter
• 金额: $ 7.43万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8357425
• 关键词: AIDS/HIV problem; Actins; Biological Assay; Capsid; Cell membrane; Cells; Complex; Cytoskeleton; Funding; Gagging; Genetic; Glycoproteins; Grant; Hour; Life; Microtubules; Minor; Motor; Movement; National Center for Research Resources; Nocodazole; Pathway interactions; Pericentriolar Regions; Physiologic pulse; Primates; Principal Investigator; Production; Recovery; Research; Research Infrastructure; Resources; Role; Site; Source; Tubular formation; Type D Retrovirus; United States National Institutes of Health; Viral; Virion; Virus; anterograde transport; cellular imaging; cost; dynein light chain; retrograde transport; trafficking

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Objectives: define (1) M-PMV Gag interactions with the cytoskeletal motor machinery, (2) role of viral and cellular components in transporting capsids from their assembly site to the plasma membrane, (3) capsid interactions with the cellular ESCRT machinery, and (4) the myristyl switch mechanism involved in budding. Efficient transport of pre-assembled capsids to the cell membrane requires a functional vesicular trafficking pathway and envelope (Env) glycoprotein. Transport may occur along a tubular-vesicular pathway induced by Env or another viral component, or these interactions may initiate Gag transport along other trafficking networks. We show Gag interacts in a CTRS-dependent manner with cellular Tctex-1, a light chain of the dynein motor complex, suggesting a microtubule-dependent retrograde transport to the pericentriolar region for capsid assembly. Microtubules might also explain anterograde transport of pre-assembled capsids to the plasma membrane. In M-PMV-infected CMMT cells, pulse-chase assays revealed a nocodazole induced 1-hour delay in virus production compared to untreated controls. Actin disruption led to a minor delay in virion release, while IF disruption had negligible effects. Live cell imaging showed that following microtubule disruption, an initial reduction in linear movement of capsids containing GFP-tagged Gag occurred, followed by a recovery of velocities. Nocodazole treatment led to a cessation in Env transport and deficient Env incorporation in released virions. Importantly, a continued albeit delayed release of virions was observed from cells lacking all three functional cytoskeleton components, suggesting a cytoskeleton-independent mode of transport. Studies are underway to alternate transport pathways. These studies could have implications for HIV/AIDS research.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 目标：定义（1）M-PMV与细胞骨架运动机械的相互作用，（2）病毒和细胞成分在将帽子从组装位点传输到质膜中的capsids中的作用，（3）与capsID相互作用，与细胞的ESCRT机械相互作用，以及（4）涉及Myristyl switch in in puss of pust offulding offulding offulding。预组装的衣壳向细胞膜有效运输需要功能性囊泡运输途径和包膜（ENV）糖蛋白。运输可能会沿着ENV或其他病毒成分引起的管状腔途径发生，或者这些相互作用可能会沿其他运输网络启动插入运输。我们以CTRS依赖性方式与Clular Tctex-1（Dynein Motor复合物的轻链）相互作用，这表明微管依赖性逆行逆行转运到capsidssbly。微管也可能解释了预组装的衣壳向质膜的顺行转运。在M-PMV感染的CMMT细胞中，与未经治疗的对照相比，脉搏疗法测定显示病毒产生1小时的延迟。肌动蛋白的破坏导致病毒粒子释放的延迟较小，而如果破坏效果可忽略不计。活细胞成像表明，在微管破坏后，发生了含有GFP标记的GAG的衣壳的线性运动的初始减少，然后恢复了速度。 Nocodazole的治疗导致了ENV运输的停止，并在释放的病毒体中置入了不足的设想。重要的是，尽管缺乏所有三个功能性细胞骨架成分的细胞，但仍延迟了病毒体的释放，这表明是细胞骨架非依赖性的转运模式。研究正在进行替代运输途径。这些研究可能对艾滋病毒/艾滋病研究有影响。