SAM68 AND HIV REPLICATION

SAM68 和 HIV 复制

• 批准号: 6214331
• 负责人: RAGHAVENDAR R THIPPARTHI
• 金额: $ 2.95万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-07-01 至 2000-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6214331
• 关键词: T lymphocyte; antibody; antisense nucleic acid; gene expression; genetic transduction; human immunodeficiency virus; nucleoproteins; protein structure function; ribozymes; tissue /cell culture; virus replication

DESCRIPTION (from applicant's abstract): Recent studies from the investigator's laboratory have identified a cellular nuclear protein, Sam68, which specifically interacts with RRE and can substitute for as well as synergize with Rev in RRE mediated gene expression and virus replication. The major goals of this proposal are to functionally characterize Sam68 protein in RRE-mediated transactivation and determine the relevance of Sam68 in HIV replication. The specific aims are: 1) to functionally characterize the interactions of Sam68 with Rev and/or RRE RNA in RRE-mediated gene expression using truncated and site-directed mutants to map domains of Sam68 and its binding sites on RRE as well as Rev that are functionally involved in RRE-mediated gene expression and virus replication; 2) to investigate the functional relevance of Sam68 in HIV replication by inactivation of Sam68 genes using ribozymes, antisense, and anti- Sam68 antibodies. This Aim will also generate an in vitro model for the enhancement of HIV replication in NIH 3T3 cells by Sam68; 3) to assess the long-term protection of primary cells expressing transdominant mutants of Sam68 from HIV infection by transducing T lymphocytes and primary cells with a retroviral vector encoding transdominant negative mutants of Sam68 and assessing long-term protection of these cells from HIV-1 infection. The effect of Sam68 on HIV mutants that are resistant to Rev M10 will also be assessed. These studies may allow the development of novel anti-viral agents.

描述（来自申请人的摘要）：研究者的最新研究 实验室鉴定出一种细胞核蛋白 Sam68， 与 RRE 特异性相互作用，可以替代或协同作用 Rev在RRE介导的基因表达和病毒复制中。主要目标 该提案的目的是在 RRE 介导中对 Sam68 蛋白进行功能表征 反式激活并确定 Sam68 在 HIV 复制中的相关性。这 具体目标是：1）从功能上表征 Sam68 的相互作用 在RRE介导的基因表达中使用Rev和/或RRE RNA，使用截短和 定点突变体将 Sam68 的结构域及其 RRE 上的结合位点映射为 以及 Rev 在功能上参与 RRE 介导的基因表达和 病毒复制； 2) 研究Sam68在HIV中的功能相关性 使用核酶、反义链和反义链使 Sam68 基因失活来进行复制 抗Sam68抗体。该目标还将生成一个体外模型 Sam68 增强 NIH 3T3 细胞中的 HIV 复制； 3）评估 对表达 Sam68 反显性突变体的原代细胞的长期保护 通过转导 T 淋巴细胞和原代细胞来避免 HIV 感染 编码Sam68和的转显性负突变体的逆转录病毒载体 评估这些细胞免受 HIV-1 感染的长期保护。效果 还将评估 Sam68 对 Rev M10 具有抗性的 HIV 突变体的作用。 这些研究可能有助于开发新型抗病毒药物。