Understanding how HIV-1 Vpu and HIV-2 Env stimulate virus release

了解 HIV-1 Vpu 和 HIV-2 Env 如何刺激病毒释放

• 批准号: 8110215
• 负责人: Paula M Cannon
• 金额: $ 8.45万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-18 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8110215
• 关键词: Acquired Immunodeficiency Syndrome; Affect; Anti-HIV Agents; Anti-HIV Therapy; Biochemical; Biology; Cell Line; Cell physiology; Cell surface; Cells; Cellular Structures; Code; Complementary DNA; Complex; Cytoplasmic Tail; Data; Employee Strikes; Enhancers; Ensure; Gagging; Gene Expression; Goals; HIV; HIV Infections; HIV-1; HIV-2; Hand; Hela Cells; Homologous Gene; Human; Human Cloning; Immunofluorescence Immunologic; Ion Channel; Knowledge; Lentivirus Vector; Libraries; Location; Lymphocyte; Macaca; Maps; Mediating; Membrane; Microarray Analysis; Modeling; Molecular; Murine leukemia virus; Open Reading Frames; Pathogenicity; Pathology; Pathway interactions; Patients; Phenotype; Play; Population; Primate Lentiviruses; Principal Investigator; Process; Production; Property; Proteins; Public Health; Recruitment Activity; Research; Retroviral Vector; Retroviridae; Role; SIV; Screening procedure; Site; Small Interfering RNA; Specificity; Suggestion; Testing; Therapeutic Intervention; Time; Viral; Viral Proteins; Virus; Virus Assembly; base; cell type; cellular targeting; drug development; env Gene Products; env Glycoproteins; gag Gene Products; in vivo; macrophage; murine retroviral vector; particle; programs; trafficking; vector genome; vpu Protein

Primate lentiviruses code for a conserved function that is necessary for the efficient production of viral particles and in vivo pathogenicity. For HIV-1 this is carried out by the accessory protein Vpu, while for HIV-2, the function has been mapped to the Env glycoprotein. In addition, our studies with the simple retrovirus, murine leukemia virus (MLV), have identified a similar activity in its Env protein. We have found that all three of these enhancers of virus release (EVRs) can boost the production of heterologous retroviruses, suggesting that they act on the general cellular pathways involved in retrovirus budding. Analysis of EVR-responsive and non-responsive cell lines has led to the suggestion that human cells contain a natural restriction factor that suppresses retrovirus budding and which the EVRs have evolved to counteract. Therefore, it is our central hypothesis that EVR function is a conserved and important activity for retroviruses in general. Although the EVR activity of Vpu was first described in 1989, the mechanism whereby it enhances virus release is still unknown and represents a significant gap in our knowledge of HIV biology. Even less is understood about the HIV-2 Env activity, which shows both similarities and striking differences with Vpu. The research plan outlined in this proposal will take a comprehensive approach to studying retroviral EVRs, both in order to identify the functional domains of the viral proteins involved, and to understand the host cell processes that are the targets of their activity. Using the three different EVRs that we have identified, we will precisely define the functional domains within these proteins and assess whether they have the same cellular target and activity, or whether a common EVR phenotype is achieved through different pathways. We will also take advantage of a non-Vpu-responsive HeLa cell line that we have identified, which retains the ability to respond to the HIV-2 Env EVR. Preliminary studies of this cell lines are already suggesting a mechanism for Vpu `s activity. We will directly examine how EVRs affect virus assembly and budding and their influence on cellular trafficking pathways, using a combination of microarray, biochemical and immunofluorescence approaches. Finally, we will attempt to clone the human cell restriction factor using approaches based on the microarray analysis of complex cDNA and siRNA libraries. Relevance of research to public health: HIV infection and AIDS continue to be a major public health problem, with anti-HIV drugs only available against limited viral targets. The HIV-1 Vpu protein is essential for HIV pathogenicity but its mechanism of action is unclear. It is our goal to investigate the mechanism behind this important HIV function and thereby identify new targets for drug development.

灵长类动病毒代码用于有效生产病毒所必需的保守功能 颗粒和体内致病性。对于HIV-1，这是由配件蛋白VPU进行的，而对于HIV-2， 该功能已映射到ENV糖蛋白。此外，我们对简单逆转录病毒的研究， 鼠白血病病毒（MLV）在其ENV蛋白中发现了相似的活性。我们发现这三个 这些病毒释放（EVR）的增强子可以提高异源逆转录病毒的产生，这表明 它们作用于逆转录病毒萌芽的一般细胞途径。 EVR响应性分析和 无反应性细胞系导致了这样的建议，即人类细胞包含自然限制因素，即 抑制逆转录病毒萌芽，EVR已进化为抵消。因此，这是我们的中心 假设EVR功能是逆转录病毒的保守和重要活性。 尽管VPU的EVR活性首次在1989年进行了描述，但它增强了病毒的机制 释放仍然未知，代表了我们对HIV生物学知识的显着差距。更少的是 了解HIV-2 ENV活动，该活动显示出与VPU的相似性和引人注目的差异。这 该提案中概述的研究计划将采取一种全面的方法来研究逆转录病毒EVRS 为了识别涉及病毒蛋白的功能域，并了解宿主细胞过程 那是他们活动的目标。使用我们确定的三种不同的EVR，我们将精确 定义这些蛋白质中的功能域，并评估它们是否具有相同的细胞靶标和 活性，或是否通过不同的途径实现常见的EVR表型。我们也会接受 我们已经确定的非VPU响应HELA细胞系的优势，它保留了响应能力 致HIV-2 ENV EVR。该细胞系的初步研究已经提出了一种VPU的机制 活动。我们将直接研究EVR如何影响病毒组装和萌芽及其对细胞的影响 运输途径，结合了微阵列，生化和免疫荧光方法。 最后，我们将尝试使用基于微阵列的方法来克隆人类细胞限制因子 复杂cDNA和siRNA文库的分析。 研究与公共卫生的相关性：艾滋病毒感染和艾滋病仍然是一个主要的公共卫生问题， 抗HIV药物仅针对有限的病毒靶标使用。 HIV-1 VPU蛋白对于HIV至关重要 致病性，但其作用机理尚不清楚。我们的目标是调查此机制 重要的HIV功能，从而确定了药物开发的新目标。

项目成果

Anti-tetherin activities in Vpu-expressing primate lentiviruses.

• DOI: 10.1186/1742-4690-7-13
• 发表时间: 2010-02-18
• 期刊: Retrovirology
• 影响因子: 3.3
• 作者: Yang SJ;Lopez LA;Hauser H;Exline CM;Haworth KG;Cannon PM
• 通讯作者: Cannon PM

Determinants in HIV-2 Env and tetherin required for functional interaction.

• DOI: 10.1186/s12977-015-0194-0
• 发表时间: 2015-08-07
• 期刊: Retrovirology
• 影响因子: 3.3
• 作者: Exline CM;Yang SJ;Haworth KG;Rengarajan S;Lopez LA;Droniou ME;Seclen E;Cannon PM
• 通讯作者: Cannon PM

Lack of adaptation to human tetherin in HIV-1 group O and P.

• DOI: 10.1186/1742-4690-8-78
• 发表时间: 2011-09-28
• 期刊: Retrovirology
• 影响因子: 3.3
• 作者: Yang SJ;Lopez LA;Exline CM;Haworth KG;Cannon PM
• 通讯作者: Cannon PM