MOLECULAR VIROLOGY OF THE FELINE IMMUNODEFICIENCY VIRUS PROTEASE

这项研究的目的是利用猫慢病毒， FIV，作为一个测试策略的系统，旨在合理设计 对慢病毒感染有效的抗病毒药物。 的相似性 FIV和HIV在生命周期、基因组组成和疾病病因方面的差异 使猫成为一个特别相关的模型。 在 此外，猫的相对经济性、安全性和可操作性 系统使其更适合详细的测试比任何 灵长类系统 研究将围绕FIV的特征 蛋白酶（PR）和FIV内被这种关键蛋白酶切割的位点 酵素 将制备真核和原核表达克隆 为了1）确定PR切割位点; 2）建立 切割; 3）评估天然和合成PR对 天然底物;和4）监测药物产生的影响 项目1-4关于天然底物的Pr裂解。 生成的数据将 用于项目4中的合成底物制备;药物设计 项目1、2和3;用于与HIV PR比较切割特异性; 并帮助建立逆转录病毒PR的共识规则， 乳沟 该项目还将负责编制 基于预测的酶之间的相互作用， 在项目1-4中确定的。 我们还将执行所有 该计划的病毒学方面，这将涉及在体外测试 对选定的药物进行毒性和抗病毒效果的测试。 的结果 这些研究将完成发育循环的锚阶段， 为设计战略提供支持和反馈， 整体方案。

The purpose of the proposed research is to utilize the feline lentivirus, FIV, as a system for testing strategies aimed at the rational design of antivirals efficacious against lentivirus infections. The similarities between FIV and HIV in life cycle, genomic make-up, and etiology of disease resulting from infection make the cat a particularly relevant model. In addition, the relative economy, safety, and manipulability of the feline system make it much more amenable to detailed testing than any of the primate systems. Studies will center around the characterization of FIV protease (PR) and the sites within FIV that are cleaved by this critical enzyme. Both eucaryotic and procaryotic expression clones will be prepared in order to 1) determine PR cleavage sites; 2) establish a hierarchy of cleavage; 3) assess cleavage rates of the natural and synthetic PRs against native substrates; and 4) monitor the influence of drugs arising from Projects 1-4 on Pr cleavage of natural substrates. The data generated will be used for synthetic substrate preparation in Project 4; drug design by projects 1, 2 and 3; for comparison of cleavage specificities with HIV PR; and to aid in the establishment of consensus rules for retroviral PR cleavage. This project will also be responsible for the preparation of site-directed mutants of PR, based on predicted interactions between enzyme and drug, established in Projects 1-4. We will also carry out all virological aspects of the program, which will involve the in vitro testing of selected drugs for both toxicity and antiviral efficacy. The results of these studies will fulfill the anchor phase of the developmental loop, providing support and feedback for design strategies arising from the overall program.