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.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 目的：定义（1）M-PMV Gag与细胞骨架运动机制的相互作用，（2）病毒和细胞组分在将衣壳从其组装位点转运到质膜中的作用，（3）衣壳与细胞ESCRT机制的相互作用，和（4）涉及出芽的肉豆蔻基开关机制。预组装衣壳向细胞膜的有效运输需要功能性囊泡运输途径和包膜（Env）糖蛋白。运输可能沿着由Env或另一种病毒组分诱导的小管-囊泡途径沿着发生，或者这些相互作用可能启动Gag沿着沿着其他运输网络的运输。我们表明，加格相互作用的CTRS依赖的方式与细胞Tctex-1，轻链的动力蛋白马达复合物，这表明微管依赖的逆行运输到pericentriolar区域的衣壳组装。微管也可能解释顺行运输的预先组装的衣壳质膜。在M-PMV感染的CMMT细胞中，脉冲追踪试验显示，与未处理的对照相比，诺考达唑诱导的病毒产生延迟1小时。肌动蛋白破坏导致病毒体释放的轻微延迟，而IF破坏的影响可以忽略不计。活细胞成像显示，微管破坏后，含有GFP标记的Gag的衣壳的线性运动发生初始减少，随后恢复速度。诺考达唑处理导致Env转运停止和释放的病毒体中Env掺入不足。重要的是，从缺乏所有三种功能性细胞骨架组分的细胞中观察到病毒粒子的持续但延迟的释放，表明了不依赖于细胞骨架的转运模式。目前正在研究替代运输途径。这些研究可能对艾滋病毒/艾滋病研究产生影响。