Alleviation of HIV-1 Rev Block in Astrocytes by an E3 Ubiquitin Ligase, NKLAM

E3 泛素连接酶 NKLAM 减轻星形胶质细胞中的 HIV-1 Rev 阻断

• 批准号: 8301516
• 负责人: RAGHAVENDAR R THIPPARTHI
• 金额: $ 19万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-15 至 2014-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8301516
• 关键词: AIDS Dementia Complex; Acquired Immunodeficiency Syndrome; Asses; Astrocytes; Binding; Brain; Cell Nucleus; Data; Development; Environment; Enzymes; Epidemic; Gene Expression; Genetic Transcription; Goals; HIV; HIV-1; Highly Active Antiretroviral Therapy; Human; Infection; Intervention; Knowledge; Laboratories; Lead; Life Cycle Stages; Lytic; Mediating; Methodology; Microscopic; Mission; Molecular; Outcome; Pathway interactions; Patients; Post-Transcriptional Regulation; Prevention; Production; Proteins; Public Health; Publishing; RNA; Reporting; Research; Resistance; Role; SRC-associated p68 protein; Testing; Therapeutic; Time; Transactivation; Ubiquitination; Vaccines; Variant; Viral; Work; base; cellular targeting; combat; experience; innovation; insight; mRNA Export; mortality; novel strategies; novel therapeutics; ubiquitin-protein ligase

DESCRIPTION (provided by applicant): There is a fundamental gap in our understanding of how HIV-1 maintains latency in astrocytes. This gap denotes a significant problem, because, unless this gap is filled, understanding the molecular mechanisms of HIV-1 latency leading to HIV-associated dementia (HAD) cannot be achieved. Furthermore, the knowledge required to develop novel therapeutic strategies for HAD patients will remain largely unknown. The long-term goal is to understand the molecular mechanisms that contribute to HIV-1 latency in the brain and develop a therapeutic strategy to combat HIV. The objective of this application is to determine the role of Natural Killer Lytic-Associated Molecule (NKLAM) in the post-transcriptional regulation of HIV-1 gene expression. The central hypothesis of this application is that NKLAM synergizes with Sam68 and alleviates a post-transcriptional block to HIV-1 production in astrocytes. The hypothesis has been formulated from the strong preliminary data, demonstrating that NKLAM interacts with Sam68, which in turn, regulates HIV-1 Rev function in astrocytes. The rationale for the proposed project is that, once the role of NKLAM in the HIV-1 life cycle is understood, its production can be modulated in such a way that impacts HIV-1 production in the prevention and treatment of AIDS, including HAD. Guided by the preliminary data, the central hypothesis will be tested to accomplish the objective of this application by pursuing the following two specific aims: 1) Determine the functional significance of NKLAM in HIV-1 replication. The working hypothesis is that NKLAM relieves a post-transcriptional block to HIV-1 production in astrocytes. Under this aim, stable NKLAM expressing astrocytes will be created and assessed for the alleviation of Rev block and HIV-1 production by examining viral mRNA export. 2) Investigate the mechanism of action of NKLAM in Sam68/RRE-mediated transactivation. The working hypothesis is that NKLAM must act in the nucleus to synergize with Sam68. Employing mutational, microscopic and ubiquitination analyses, the mechanism of action of NKLAM in Sam68/Rev/RRE function will be determined. The proposed work is innovative, because, so far, no E3 ubiquitin ligase(s), specifically NKLAM, has been implicated in the post-transcriptional regulation of HIV-1 gene expression. Most importantly, to date, there are no published reports implicating, any E3 ligase(s), particularly, NKLAM- mediated Sam68 ubiquitination in alleviating the Rev block in astrocytes. The proposed research is significant because it is expected to provide the knowledge needed to develop novel strategies to eliminate HIV-1 reservoirs and combat HAD.

描述（由申请人提供）：我们对HIV-1如何保持星形胶质细胞延迟的理解存在基本差距。该差距表示一个重大的问题，因为除非填补了这个差距，否则无法实现HIV-1潜伏期的分子机制，导致与HIV相关的痴呆症（HAT）无法实现。此外，制定新的患者治疗策略所需的知识将在很大程度上未知。长期目标是了解有助于大脑中HIV-1潜伏期的分子机制，并制定一种治疗策略来对抗HIV。该应用的目的是确定自然杀伤质裂解相关分子（NKLAM）在HIV-1基因表达的转录后调节中的作用。该应用的中心假设是NKLAM与SAM68协同作用，并减轻了星形胶质细胞中HIV-1产生的转录后阻滞。该假设是从强大的初步数据中提出的，表明NKLAM与SAM68相互作用，这反过来又调节了星形胶质细胞中的HIV-1 Rev功能。拟议项目的基本原理是，一旦NKLAM在HIV-1生命周期中的作用被理解，其产量就可以调节，以影响HIV-1产生在预防和治疗艾滋病（包括HAT）中的方式。在初步数据的指导下，将通过追求以下两个特定目的来测试中心假设以实现本应用的目标：1）确定NKLAM在HIV-1复制中的功能意义。工作的假设是，NKLAM将转录后阻滞减轻了星形胶质细胞中的HIV-1产生。在此目标下，将通过检查病毒mRNA出口来创建和评估稳定的NKLAM表达星形胶质细胞，以减轻Rev Block和HIV-1的产生。 2）研究NKLAM在SAM68/RRE介导的反式激活中的作用机理。工作假设是Nklam必须在核中起作用才能与SAM68协同作用。采用突变，微观和泛素化分析，将确定NKLAM在SAM68/REV/RRE功能中的作用机理。提出的工作具有创新性，因为到目前为止，尚无E3泛素连接酶，特别是NKLAM，与HIV-1基因表达的转录后调节有关。最重要的是，迄今为止，尚无公开的报告，任何E3连接酶，尤其是Nklam介导的SAM68泛素化，以减轻星形胶质细胞的转速阻滞。拟议的研究很重要，因为有望提供开发新型策略来消除HIV-1储层和战斗所需的知识。