TAT ASSOCIATED KINASE REGULATION OF HIV GENE EXPRESSION

HIV 基因表达的 TAT 相关激酶调节

• 批准号: 2673092
• 负责人: Richard B Gaynor
• 金额: $ 21.59万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-01 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2673092
• 关键词: DNA directed RNA polymerase; adenosinetriphosphatase; enzyme activity; gene expression; helicase; host organism interaction; human immunodeficiency virus 1; human tissue; phosphotransferases; transcription factor; virus protein

DESCRIPTION: (Adapted From Investigator's Abstract) Tat stimulates gene expression by increasing the ability of RNA polymerase II to efficiently undergo transcriptional elongation. Recent studies suggest that tat can bind a cellular kinase that phosphorylates the C-terminal domain (CTD) of RNA polymerase II. The investigator has purified the kinase complex and demonstrated that it contains novel components as well as previously characterized components of the multi-protein transcription factor, TF II H. Wildtype, but not mutant tat proteins, greatly enhance the ability of the kinase complex to phosphorylate the RNA polymerase II CTD and addition of the kinase to in vitro transcription assays with the HIV-1 LTR markedly stimulated tat activation, suggesting that tat-associated kinase complex is a critical cellular factor involved in tat activation of HIV-1. The investigator proposes to examine the mechanism by which tat association with a kinase complex increases HIV-1 gene expression. Specific aims include: Determine which cellular components of the kinase complex are able to directly associate with tat. Assay the function of novel components of the tat-associated complex using both in vitro and in vivo assays. Determine whether tat facilitates association of the kinase complex with RNA polymerase II and whether tat alters enzymatic activities of TF II H. Develop a reconstituted in vitro transcription system in which tat activation is dependent on the addition of the tat-associated kinase and further to determine the role of this kinase on regulating in vivo HIV-1 gene expression.

描述：（改编自研究者摘要）Tat 刺激基因 通过增加 RNA 聚合酶 II 的能力来有效地表达 经历转录延伸。 最近的研究表明，tat 可以 结合磷酸化 C 端结构域 (CTD) 的细胞激酶 RNA聚合酶II。 研究人员纯化了激酶复合物并 证明它包含新颖的成分以及以前的 多蛋白转录因子 TF II H 的特征成分。 野生型而非突变型 tat 蛋白大大增强了 磷酸化 RNA 聚合酶 II CTD 的激酶复合物并添加 激酶与 HIV-1 LTR 的体外转录测定显着 刺激 tat 激活，表明 tat 相关激酶复合物是 参与 HIV-1 的 tat 激活的关键细胞因子。 这 研究者建议研究tat与 激酶复合物可增加 HIV-1 基因表达。 具体目标包括： 确定激酶复合物的哪些细胞成分能够 直接与 tat 相关联。 测定新成分的功能 tat 相关复合物使用体外和体内测定。 决定 tat是否促进激酶复合物与RNA的结合 聚合酶 II 以及 tat 是否改变 TF II H 的酶活性。 开发重建的体外转录系统，其中 tat 激活依赖于tat相关激酶的添加 进一步确定该激酶在体内调节 HIV-1 中的作用 基因表达。