Clinical Implications of HIV-1 Rev-Rev Response Element Functional Activity Variation

HIV的生命周期需要含有内含子的病毒mRNA从细胞核输出到细胞质。这 这一过程通常受到细胞的限制，但艾滋病毒通过与病毒的相互作用克服了这一限制。 蛋白质Rev和RNA二级结构Rev-反应元件（RRE），在含有内含子的 病毒转录本在所取的病毒中观察到Rev-RRE轴功能活性的变化 但这种变异的临床意义及其因果机制并不清楚， 明白了以前对相关慢病毒的研究清楚地表明，Rev-RRE活性变化在慢病毒中起着重要作用。 在发病机制中的作用，强烈表明该系统在HIV感染中同样重要。是 假设Rev-RRE调节轴起变阻器的作用，允许病毒适应 不同的免疫环境。在这个K 08指导临床科学家研究职业发展奖 弗吉尼亚大学传染病研究员帕特里克杰克逊博士提出， 进行了一系列研究，以阐明Rev-RRE系统在HIV传播和病毒感染中的作用。 延迟。首先，使用一种新的Rev-RRE功能活性的高通量慢病毒载体测定， HIV传播中的这一轴将通过比较传播和循环中的Rev-RRE活性来检查。 病毒准种其次，比较潜伏前病毒和循环病毒的Rev-RRE活性。 将检测高活性和低活性Rev-RRE对，以确定它们是否导致病毒复制动力学的差异 和病毒蛋白质的产生。这种现象可能是“踢死”战略失败的原因， 治愈艾滋病最后，将进行结构域交换实验以鉴定 Rev和RRE功能活动。将利用天然存在的和嵌合的Rev和RRE序列 以确定关键的残留物和结构。这项研究对开发改良的 艾滋病毒传播的药物预防和“踢和杀”艾滋病毒治疗的改进。这 研究将在弗吉尼亚大学进行，由大卫雷科什博士和玛丽博士指导- 路易丝·哈马舍尔德有着良好的训练成功记录。杰克逊医生提议 发展计划包括体验式学习;分子生物学，生物信息学， 和赠款写作;和结构化的研究和专业指导。他提出了具体的时间轴， 在国际会议上和在同行审查的出版物中介绍成果， 未来的补助金申请这项计划旨在使他成为一名独立的研究人员， 在艾滋病毒学和一套技能，其中包括一套广泛的科学技术。

The HIV life cycle requires the export of intron-containing viral mRNAs from the nucleus to the cytoplasm. This process is ordinarily restricted by the cell, but HIV overcomes this check through the interaction of a viral protein, Rev, and an RNA secondary structure, the Rev-Response Element (RRE), found on intron-containing viral transcripts. Variation in the functional activity of the Rev-RRE axis has been observed in viruses taken from patients infected with HIV, but the clinical significance of that variation and its causal mechanism is not clearly understood. Previous studies of a related lentivirus clearly show that Rev-RRE activity variation plays a role in pathogenesis, strongly suggesting that this system is similarly important in HIV infection. It is hypothesized that the Rev-RRE regulatory axis functions as a rheostat which permits viral adaptation to different immune environments. In this K08 Mentored Clinical Scientist Research Career Development Award application, Dr. Patrick Jackson, a fellow in Infectious Diseases at the University of Virginia, proposes to perform a series of studies to illuminate the role of the Rev-RRE system in HIV transmission and in viral latency. First, using a novel high-throughput lentiviral vector assay of Rev-RRE functional activity, the role of this axis in HIV transmission will be examined by comparing Rev-RRE activity in transmitted and circulating viral quasispecies. Second, the Rev-RRE activity of latent proviruses and circulating viruses will be compared. High and low activity Rev-RRE pairs will be tested to see if they cause differences in viral replication kinetics and the production of viral proteins. This phenomenon could underlie the failure of “kick and kill” strategies to cure HIV. Finally, domain swapping experiments will be performed to identify the sequence determinants of both Rev and RRE functional activity. Naturally occurring and chimeric Rev and RRE sequences will be utilized to identify key residues and structures. This research has direct implications for the development of improved pharmacologic prophylaxis for HIV transmission and for improvements in “kick and kill” HIV cures. This research will be conducted at the University of Virginia under the mentorship of Drs. David Rekosh and Marie- Louise Hammarskjold who have a strong track record of training success. Dr. Jackson proposes a career development plan consisting of experiential learning; formal course work in molecular biology, bioinformatics, and grant writing; and structured research and professional mentorship. He proposes a specific timeline for presentation of results at international conferences and in peer reviewed publications, as well as a timeline for future grant applications. This program is designed to situate him as an independent researcher with expertise in HIV virology and a skill set which encompasses a broad set of scientific techniques.