EFFECTS OF STRESS/STRESS RESPONSE GENES ON REGULATION OF HIV-1 GENE EXPRESSION

用DNA损伤剂治疗人类细胞，如紫外线辐射， 会导致潜伏病毒的激活和病毒的增加 制作。这可能对疾病的进展有重要的影响。 从环境来源和在治疗艾滋病毒-1感染期间 使用细胞毒剂的患者。我们在基因毒性应激方面的专业知识 反应已被应用于艾滋病毒-1反应的研究。这 该项目是由NIH针对艾滋病的抗病毒计划资助的 对于91财年和92财年仅对93财年的研究员提供部分工资支持的情况： 从那时起，它就得到了DTP资金的支持。最初，我们 集中在细胞毒剂对稳定整合的HIV- 启动子-CAT构建。我们已经发现，体内的一种或多种细胞因子 介质可以影响DNA损伤的反应性；此外，我们还 发现紫外线照射细胞产生的一种可溶性因子(S)可以 激活稳定整合的HIV-CAT反式。因为紫外线照射可以 诱导一种细胞TAT样因子，我们对细胞因子的研究 参与HIV-1调控的主要是带有TAT-1的蛋白质- 类似的属性，我们已经通过西北大学的印迹鉴定 探讨全核中多种TAR RNA结合蛋白(TBP) 萃取物。TBP的水平在不同的细胞系中是不同的， 在许多情况下都是DNA损伤的诱因。使用这种方法，我们有 已知的焦油结合蛋白的数量增加了一倍多，并显示出 这是TBP对压力做出反应的第一个演示。我们计划进一步 描述这些TBP及其对遗传毒性应激的反应，包括 细胞因子在其激活中的作用。将开发各种方法来 纯化这些DNA损伤诱导的RNA结合蛋白并最终 基于部分蛋白质序列测定分离cDNA克隆。 对这些蛋白质的鉴定可能会提供重要的见解 HIV-1的调控和HIV-1感染细胞对 细胞毒剂。

Treatment of human cells with DNA-damaging agents, such as UV radiation, can lead to the activation of latent virus and to increased virus production. This may have important implications in disease progression both from environmental sources and during treatment of HIV-1-infected patients with cytotoxic agents. Our expertise in genotoxic stress responses has been applied to the study of responses in HIV-1. This project was funded by the NIH Intramural AIDS Targeted Antiviral Program for FY91 and FY92 with only partial salary support for a fellow in FY93: since that time, it has been supported by DTP funding. Initially, we focused on the induction by cytotoxic agents of stably-integrated HIV- promoter-CAT constructs. We have found that one or more cytokines in the medium can affect DNA-damage responsiveness; in addition, we have also found that a soluble factor(s) produced by UV-irradiated cells can activate stably-integrated HIV-CAT in trans. Since UV-irradiation can induce a cellular Tat-like factor, our search for cellular factors involved in HIV-1 regulation has focused primarily on proteins with Tat- like properties, and we have identified by a Northwestern blotting approach a variety of TAR RNA-binding proteins (TBP) in whole nuclear extracts. The levels of TBP were found to vary in different cell lines, and were DNA-damage inducible in many cases. With this approach, we have more than doubled the number of known TAR-binding proteins and have shown the first demonstration of a TBP response to stress. We plan to further characterize these TBP and their responses to genotoxic stress including the role of cytokines in their activation. Approaches will be developed to purify these DNA-damage-inducible RNA-binding proteins and to ultimately isolate cDNA clones based on partial protein sequence determinations. Identification of these proteins will probably provide important insights into the regulation of HIV-1 and the responses of HIV-1-infected cells to cytotoxic agents.