Identification of novel anti-HIV inhibitors based on Vif-E3 activity

基于 Vif-E3 活性鉴定新型抗 HIV 抑制剂

• 批准号: 8713917
• 负责人: Richard B. Markham
• 金额: $ 20.25万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-05 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8713917
• 关键词: 26S proteasome; Acquired Immunodeficiency Syndrome; Antiviral Agents; Biological Assay; Biological Models; CD4 Positive T Lymphocytes; Cell Line; Cells; Core-Binding Factor; Cytidine Deaminase; Cytosine; Development; Goals; HIV; HIV-1; Host Defense; Human; In Vitro; Intervention; Knowledge; Libraries; Mammalian Cell; Mediating; Modification; Molecular; Polyubiquitination; Proteins; Research; Reverse Transcription; System; Testing; Toxic effect; Uracil; Viral; Virion; Virus; Virus Diseases; Virus Inhibitors; antiretroviral therapy; base; counterscreen; design; effective intervention; genetic regulatory protein; high throughput screening; human CEM15 protein; inhibitor/antagonist; macrophage; novel; novel strategies; public health relevance; screening; small molecule; ubiquitin-protein ligase; viral DNA; yeast two hybrid system

DESCRIPTION (provided by applicant): Human cytidine deaminases APOBEC3 (A3) proteins are potent host defenses against HIV. These antiviral proteins induce lethal modification of cytosines to uracils in newly synthesized minus-strand viral DNA, resulting in abortive viral infection. HIV must overcome these host cellular defenses for successful viral replication. HIV-1 encodes a protein, Vif, which suppresses the antiviral effects of A3 proteins by targeting them for degradation through the 26S proteasome. Vif hijacks cellular Cullin5 (Cul5), ElonginB, and ElonginC to form a viral E3 ubiquitin ligase that targets A3G for polyubiquitination and degradation. Thus, identification of novel strategies to preserve the antiviral functions of A3 is n exciting new target for antiretroviral therapy. In this application, we propose to capitalize our expertise in HIV-1 Vif/A3 system and our new understanding of the viral evasion mechanism to develop a rapid cell-based assay for the identification of small molecule inhibitors of Vif and to further optimize and adapt the system for application to high throughput molecular screening of large compound libraries to identify molecules that inhibit HIV-1 replication. The proposed research is based on our recent discovery that CBF? is a key and unique regulator of HIV-1 Vif function. This study is expected to provide us with critical information regarding the design and development of effective intervention strategies against HIV.

描述（由申请人提供）：人胞苷脱氨酶APOBEC3 （A3）蛋白是抗HIV的有效宿主防御。这些抗病毒蛋白诱导新合成的负链病毒DNA中胞嘧啶对尿嘧啶的致命修饰，导致病毒感染流产。HIV必须克服宿主细胞的这些防御才能成功复制病毒。HIV-1编码一种名为Vif的蛋白，该蛋白通过26S蛋白酶体靶向A3蛋白降解，从而抑制A3蛋白的抗病毒作用。Vif劫持细胞Cullin5 （Cul5）、拉长素b和拉长素，形成病毒E3泛素连接酶，靶向A3G进行多泛素化和降解。因此，发现保留A3抗病毒功能的新策略是抗逆转录病毒治疗的一个令人兴奋的新靶点。在本申请中，我们建议利用我们在HIV-1 Vif/A3系统方面的专业知识和我们对病毒逃避机制的新理解，开发一种基于细胞的快速检测方法，用于鉴定Vif的小分子抑制剂，并进一步优化和调整该系统，用于大化合物文库的高通量分子筛选，以鉴定抑制HIV-1复制的分子。这项提议的研究是基于我们最近的发现，脑血流？是HIV-1 Vif功能的关键和独特的调节因子。这项研究有望为我们提供设计和开发有效的艾滋病干预策略的关键信息。