HIV-1 INHIBITION BY 2-5A DERIVATIVES, 2-50AS & PKR GENES

描述：(改编自申请者摘要)研究建议 重点介绍了控制HIV-1感染的两种策略，并探讨了 发生在2-5 OAS、PKR和HIV-1之间的基础生物学。第一, 抗核酸酶、无毒、生物活性的2-5A衍生物和 他们的亲脂结合物已经被合成来增加耗尽的 通过作用于HIV-1中2-5OAS阻断的远端的2-5A细胞池 被感染的细胞。这些2-5A衍生物显示对HIV-1的增强摄取 被感染的细胞。它们可能部分地通过激活核糖核酸酶来起作用 L领导的艾滋病病毒1型复制得到了抑制。这种抗病毒作用 在HIV-1复制上的2-5A衍生物中可能仅代表一个 它们在体内的作用方式的一个方面；因此，替代 将研究活动机制，包括抑制艾滋病毒-1 整合酶和HIV-1逆转录酶。2-5A的选择性 衍生品将通过测量它们对宿主细胞DNA的影响进行测试， HIV-1感染和未感染细胞的RNA和蛋白质合成。 其次，克隆了编码2-5OAS和PKR的cDNA 位于HIV-13‘LTR内的TAR结构并包装成 高效价逆转录病毒载体。一旦真核细胞稳定 转导后，这些关键的抗病毒酶将被置于 另一种控制机制。HIV-1感染后会产生TAT 蛋白质将转录激活TAR序列，导致 增加了2-5OAS和PKR的产量。随后激活这些 病毒dsRNA的酶可能最终导致对 被感染的细胞阻止病毒传播和/或潜伏期重新激活。 因此，这项提案将侧重于理解和 重组天然抗病毒宿主的机制(S) 防御途径控制着HIV-1的复制。

DESCRIPTION: (Adapted from Applicant's Abstract) The research proposed focuses on two strategies to control HIV-1 infection and explore the basic biology which occurs between 2-5OAS, PKR, and HIV-1. First, nuclease-resistant, non-toxic, biologically active 2-5A derivatives and their lipophilic conjugates have been synthesized to augment the depleted 2-5A cellular pool by acting distal to the 2-5OAS blockade in HIV-1 infected cells. These 2-5A derivatives show enhanced uptake in HIV-1 infected cells. They may act, in part, through the activation of RNase L leading to an inhibition of HIV-1 replication. This antiviral effect of the 2-5A derivatives on HIV-1 replication may only represent one aspect of their mode of action in vivo; therefore, alternative mechanisms of activity will be examined, including inhibition of HIV-1 integrase and HIV-1 reverse transcriptase. The selectivity of the 2-5A derivatives will be tested by measuring their effect on host cell DNA, RNA and protein synthesis in HIV-1 infected and uninfected cells. Second, cDNAs encoding 2-5OAS and PKR have been cloned under the control of the TAR structure located within the HIV-1 3' LTR and packaged into high titer retroviral vectors. Once eukaryotic cells are stably transduced, these key antiviral enzymes will have been placed under this alternate control mechanism. Upon infection, the HIV-1 produced Tat protein will transcriptionally activate the TAR sequence leading to the increased production of 2-5OAS and PKR. Subsequent activation of these enzymes by viral dsRNA may ultimately lead to the destruction of the infected cells preventing viral spread and/or latency reactivation. This proposal will therefore focus upon the understanding and restructuring of the mechanism(s) by which the natural antiviral host defense pathways control HIV-1 replication.