Dynamic of Retroviruses Assembly and Budding

逆转录病毒组装和出芽动态

• 批准号: 7838566
• 负责人: Nolwenn monique Jouvenet
• 金额: $ 9.27万
• 依托单位: AARON DIAMOND AIDS RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7838566
• 关键词: Acquired Immunodeficiency Syndrome; Address; Affect; African Swine Fever Virus; Animals; Antiviral Agents; Area; Binding; Biological; Biological Assay; Biology; Biophysics; Calculi; Cell membrane; Cells; Cellular biology; Collaborations; Complex; Cytoplasm; DNA Viruses; Data; Development; Diamond; Doctor of Philosophy; England; Event; Gagging; Genome; Goals; Grant; HIV; HIV-1; Head; Health; Imaging Techniques; Immunology; Individual; Infection; Institutes; Intracellular Transport; Laboratories; Laboratory Study; Life; Life Cycle Stages; Mentors; Microscopic; Names; New York; Nuclear Import; Paper; Pathway interactions; Phase; Play; Postdoctoral Fellow; Publications; RNA; Research; Research Personnel; Retroviridae; Role; Site; Techniques; Time; Training; Universities; Viral; Virion; Virus Diseases; Work; Writing; career; cell determination; gag Gene Products; innovation; interest; novel strategies; particle; post-doctoral training; tool; trafficking; viral RNA; virology

DESCRIPTION (provided by applicant): I completed my PhD in three years in the Department of Virology and Immunology of the Institute for Animal Health, England, where I studied the intracellular transport of African swine fever virus, a large DNA virus. My work led to the publication of 3 first-author papers. I graduated in September 2004. In January 2005, I joined Paul Bieniasz group at the Aaron Diamond AIDS Research Center, in New York, USA. As a post-doctoral fellow, I am continuing to explore the cell biology of virus infection. My current work focuses on the assembly and budding of Human immunodeficiency virus type I (HIV-1). I first established that the productive site of HIV-1 assembly was the plasma membrane. More recently, in collaboration with Sandy Simon, the head of the Cellular Biophysics laboratory at the Rockefeller University, we have developed innovative microscopic approaches to study HIV-1 particles assembly in real time, at the scale of individual particles. These approaches have allowed an unprecedented view of the genesis of individual virus particles in live cells and the determination of parameters that were inaccessible with conventional techniques. So far, my post-doctoral work led to the publication of 5 papers, including 3 first author papers. My immediate objectives are to exploit these approaches to understand the dynamic of the interaction between HIV-1 Gag proteins and key cellular factors at the site of particle assembly and release. During the K99 phase, I will spend more time in Sandy Simon laboratory, where I will be trained to develop new elaborate microscopic tools. I will pursue my work on the dynamic of HIV-1 assembly and release, using existing tools, as well as tools under development. Once the training phase of the grant will be completed, I intend to start my own group. As an independent investigator, I will continue exploiting the techniques and tools that I have developed during my post-doctoral training to pursue my studies on the cell biology of HIV-1 infection. Initially, during the R00 phase, I will focus on a particular aspect of HIV-1 replication: the cell biology of genome packaging. New approaches are definitively warranted in this field to determine, for instance, where in the cell the HIV-1 genome dimerizes, what mechanisms govern the cytoplasmic trafficking of the genome to assembly sites, where in the cell RNA first binds Gag, etc. This is also an area of research that is not studied in Paul Bieniasz laboratory, so my work as an independent investigator should not overlap with his. The R00 phase will allow me to start my own group without the burden of writing a grant in the first year and allow me to hire some staff immediately. After this grant-writing free time, my staff and I should have collected enough preliminary data to submit an R01. This first R01 should be a stepping-stone toward establishing a successful laboratory studying the cell biology of HIV-1. My long-term goal is to expand into other poorly understood areas of HIV-1 biology that interest me, such as on post-entry events. More specifically, I will focus my work on the formation, intracellular transport and nuclear import of partially disassembled viral particles, named pre- integration complexes. All cell biological aspects of these crucial steps of the HIV-1 life cycle are unknown; understanding them is likely to reveal new and fundamental cellular pathways. I am determined and highly motivated to pursue my career in the exciting field of HIV-1 cell biology. Receiving the competitive K99/R00 grant will certainly help me reach this goal. Project narrative: A better understanding of HIV-1 infection is crucial for the development of new classes of antiviral drugs. In order to dissect the mechanisms of HIV-1 assembly, we have developed an assay that allows us to visualize the formation of viral particles in real time and at the scale of single particles. We propose to use this innovative assay to study the dynamic interactions between key viral and cellular components during HIV-1 assembly and release.

描述（由申请人提供）：我在英国动物卫生研究所病毒学和免疫学系完成了三年的博士学位，在那里我研究了非洲猪瘟病毒（一种大型DNA病毒）的细胞内转运。我的工作导致发表了3篇第一作者论文。2004年9月毕业。 2005年1月，我加入了位于美国纽约的亚伦·戴蒙德艾滋病研究中心的保罗·比尼亚兹小组。作为博士后研究员，我继续探索病毒感染的细胞生物学。我目前的工作重点是人类免疫缺陷病毒I型（HIV-1）的组装和萌芽。我首先确定HIV-1装配的生产场所是质膜。最近，我们与洛克菲勒大学细胞生物物理实验室的负责人桑迪西蒙合作，开发了创新的显微镜方法，以真实的时间，在单个粒子的尺度上研究HIV-1粒子的组装。这些方法允许活细胞中单个病毒颗粒的起源和常规技术无法获得的参数的测定的前所未有的视图。到目前为止，我的博士后工作导致发表了5篇论文，其中包括3篇第一作者论文。 我的近期目标是利用这些方法来了解HIV-1 Gag蛋白和关键细胞因子在颗粒组装和释放位点之间的相互作用的动态。在K99阶段，我将花更多的时间在桑迪西蒙实验室，在那里我将接受培训，开发新的精细显微工具。我将继续我的工作对艾滋病毒-1的组装和释放的动态，使用现有的工具，以及正在开发的工具。 一旦培训阶段的赠款将完成，我打算开始我自己的小组。作为一名独立的研究者，我将继续利用我在博士后培训期间开发的技术和工具，继续研究HIV-1感染的细胞生物学。最初，在R 00阶段，我将专注于HIV-1复制的一个特定方面：基因组包装的细胞生物学。新的方法是明确保证在这一领域，以确定，例如，在细胞中的HIV-1基因组二聚化，什么机制管理细胞质运输的基因组组装网站，在细胞中的RNA首先结合Gag等，这也是一个研究领域，没有研究在保罗Bieniasz实验室，所以我的工作作为一个独立的研究者不应该与他重叠。R 00阶段将允许我开始我自己的小组，而无需在第一年编写资助的负担，并允许我立即雇用一些员工。在这段写补助金的空闲时间之后，我和我的工作人员应该已经收集了足够的初步数据来提交R 01。 第一个R 01应该是建立一个成功的研究HIV-1细胞生物学实验室的垫脚石。我的长期目标是扩展到我感兴趣的HIV-1生物学的其他鲜为人知的领域，例如入境后事件。更具体地说，我将集中我的工作的形成，细胞内运输和核输入的部分拆卸的病毒颗粒，命名为前整合复合物。HIV-1生命周期这些关键步骤的所有细胞生物学方面都是未知的;了解它们可能会揭示新的和基本的细胞途径。 我决心并高度积极地追求我的职业生涯在令人兴奋的领域艾滋病毒-1细胞生物学。获得具有竞争力的K99/R 00赠款肯定会帮助我实现这一目标。 项目叙述：更好地了解HIV-1感染对于开发新型抗病毒药物至关重要。为了剖析HIV-1组装的机制，我们开发了一种检测方法，使我们能够在真实的时间和单个颗粒的尺度上可视化病毒颗粒的形成。我们建议使用这种创新的检测方法来研究HIV-1组装和释放过程中关键病毒和细胞成分之间的动态相互作用。