HIV-2 GENOMIC RNA STRUCTURE

HIV-2 基因组 RNA 结构

• 批准号: 6628001
• 负责人: J. Stephen Lodmell
• 金额: $ 18.2万
• 依托单位: UNIVERSITY OF MONTANA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-02-15 至 2004-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6628001
• 关键词: RNA; dimer; gene deletion mutation; genetic mapping; genome; human immunodeficiency virus 2; nucleic acid sequence; nucleic acid structure; site directed mutagenesis; virus replication

The genomes of retroviruses are remarkably small considering the complexity of replicative control which they encode. To attain this level of parsimony, the genomic RNA contains not only coding information for proteins, but also functional signals for transactivation, dimerization, encapsidation, splicing, and poly A addition near the 5' and 3' extremities. Each of these events is vital for productive replication of the virus. In the research proposed here, the structure and function of the cis-element(s) within the genomic RNA that promote dimerization of two copies of the full-length genome will be studied in detail within the context of the HIV- 2 virus, a causative agent for AIDS in humans. Specifically, the signal for dimerization of genomic RNA will be identified and characterized using RNA structure probing, chemical modification interference analysis, mutagenesis, and deletion analysis. Sites identified by these experiments will be further characterized using biochemical methods and a combinatorial in vitro evolution approach to determine the stability and mechanism of the interaction. Detailed secondary/tertiary structure maps of the 5' leader region and parts of the gag coding sequence of the genomic RNA will be constructed to better understand potential interplay between functional motifs in the region. Finally, mutant viruses will be constructed with deletion or mutations in their dimerization domains to assess their effects in vivo. These studies will provide a vital structural basis for developing new anti-retroviral therapies directed toward the functional regions of the genomic RNA.

考虑到逆转录病毒编码的复制控制的复杂性，它们的基因组非常小。为了达到这种简约水平，基因组 RNA 不仅包含蛋白质的编码信息，还包含反式激活、二聚化、衣壳化、剪接以及 5' 和 3' 末端附近的聚腺苷酸添加的功能信号。这些事件中的每一个对于病毒的有效复制都至关重要。在此提出的研究中，将在 HIV-2 病毒（人类艾滋病的病原体）的背景下详细研究基因组 RNA 内促进全长基因组两个拷贝二聚化的顺式元件的结构和功能。具体来说，将使用RNA结构探测、化学修饰干扰分析、诱变和缺失分析来鉴定和表征基因组RNA二聚化的信号。这些实验确定的位点将使用生化方法和组合体外进化方法进一步表征，以确定相互作用的稳定性和机制。将构建基因组 RNA 的 5' 前导区和部分 gag 编码序列的详细二级/三级结构图，以更好地了解该区域中功能基序之间的潜在相互作用。最后，将构建二聚化结构域缺失或突变的突变病毒，以评估其体内效果。这些研究将为开发针对基因组 RNA 功能区域的新抗逆转录病毒疗法提供重要的结构基础。