FUNCTIONS AND INHIBITION OF HIV POLYMERASE

HIV聚合酶的功能和抑制

• 批准号: 2057210
• 负责人: Lorna RODRIGUEZ-RODRIGUEZ
• 金额: $ 7.93万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-05-01 至 1998-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2057210
• 关键词: RNA directed DNA polymerase; cis platinum compound; exonuclease; genetic manipulation; genetic transcription; human immunodeficiency virus; nucleic acid sequence; nucleoside triphosphate; protein structure function; reverse transcriptase inhibitors; ribonuclease III; virus genetics

Retrovirus reverse transcriptases (RT) catalyze a reaction called transfer synthesis, whereby a DNA primer is moved form one viral template to another. The reaction is essential for viral replication, and likely an important step in recombination. It is probable that this type of reaction, when it occurs between internal regions of the viral chromosome, contributes to diversity in the viral genome, improving viral survival. We intent to understand and eventually prevent internal strand transfer from occurring in vivo, since it may be one of the features that improves virulence. A strand transfer assay in vitro will be used to measure the capacity of the reverse transcriptase to transfer DNA synthesis from a natural sequence donor template to an acceptor template. The role of template sequence and secondary structure, and of RNA cleavage, in the strand transfer reaction will be tested. Effects of specific alterations in gap size produced in the RNA template, the size of specifically introduced hairpin loops, and degree of homology between donor and acceptor templates on the efficiency of strand transfer will be examined. The effect of covalent modifications to the donor RNA forming cisplatin adducts will also be determined. Clinically relevant experiments will be done determining the effects of BI-RG-587 and its analogues on processive primer elongation necessary for strand transfer. In summary, the experiments are designed to determine the characteristic of the templates, RT and reaction conditions that favor the internal strand transfer reaction.

逆转录病毒逆转录酶(RT)催化一种称为 转移合成，即DNA引物从一个病毒模板移出 给另一个人。这种反应对病毒复制是必不可少的，而且很可能 重组的重要一步。很可能这种类型的 当它发生在病毒内部区域之间的反应 染色体，有助于病毒基因组的多样性，改善病毒 生死存亡。我们打算了解并最终防止内部链 转移发生在体内，因为它可能是 提高毒力。体外链转移试验将用于 测定逆转录酶转移DNA的能力 从自然序列供体模板到受体模板的合成。 模板序列和二级结构以及RNA作用 裂解，在链转移反应中将进行测试。的效果 在RNA模板中产生的间隙大小的特定变化，大小 特别引入的发夹环，以及 供体和受体模板对链转移效率的影响 接受检查。共价修饰对供体RNA的影响 顺铂加合物的形成也将被确定。临床相关性 将进行实验，以确定BI-RG-587和其 链转移所需的连续型底物延伸的类似物。 总而言之，设计这些实验是为了确定 在模板中，RT和反应条件有利于内部 链转移反应。