GENETIC INTERACTION OF HUMAN HERPESVIRUS-6 WITH HIV-1

这个项目的目标是识别和描述人类的特征 能激活人类免疫缺陷的疱疹病毒6型基因 病毒1型(HAD)启动子和确定如何相互作用 在这两种病毒之间发生。HHV-6被认为是一个辅助因素 在感染和艾滋病的发展中，因为1)HHV-6 能有效感染许多与HIV-6相同类型的细胞 通过将DNA导入猴肾来增强细胞病变效应 细胞和人类T细胞，我们已经证明了HHV-6基因片段ZVH14 (8.7kb)，发现它们可以转化小鼠细胞和人类 角质形成细胞，可增加该指标的表达 氯霉素乙酰转移酶(ZCAT)基因与HAVE启动子连锁。 这种激活是通过Sp1(细胞因子)结合位点介导的 在HAD启动子中的存在和激活被显著降低 设计用来阻断Sp1结合位点的低聚物。为提高效率 激活时，需要多个Sp1位点。核转录因子-kB位点 HAVE启动子不是HHV-6ZVH12介导的激活所必需的。 我们对ZVH14缺失锥体的研究发现，一个小的开放 被称为B115的阅读框可能编码115个氨基酸的蛋白质， 主要参与了这一激活功能。初步 体外表达研究发现，预测的18R=18的蛋白 KDA。最近，我们已经通过位点产生了这种蛋白质的突变体- 特异性突变以确认其在这种反式激活中的作用 复制。这项研究对于更好地理解HHV- 6在HIV发病机制中的作用。该项目还将提供知识和工具 用于开发新的抗病毒疗法和安全的病毒载体 艾滋病的基因治疗。

he objective of this project is to identify and characterize the human herpesvirus-6 (HHV-6) genes that can activate the human immunodeficiency virus type-1 (HAVE) promoter and to determine how the interactions between these two viruses occur.HHV-6 has been suggested as a cofactor in HAVE infection and in the development of AIDS, since 1) HHV-6 productively infects many of the same cell types as HIV-6 lead to increased cytopathic effects.By DNA transfection into monkey kidney cells and human T cells, we have shown that a HHV-6 gene segment ZVH14 (8.7 kb), which were found to transform mouse cells and human keratinocytes, can increase the expression of the indicator chloramphenicol acetyltransferase zCAT) gene linked to the HAVE promoter. This activation was mediated through a Sp1 (cellular factor) binding site present in the HAVE promoter and activation was dramatically reduced by oligomers designed to block the Sp1 binding sites. For efficient activation, more than one Sp1 site were required. NF-kB sites of the HAVE promoter were not essential for HHV-6 ZVH12-mediated activation. Our studies with deletion cones of ZVH14 identified that a small open reading frame called B115, potentially encoding a 115-amino acid protein, was primarily involved in this transactivation function. Preliminary expression studies in vitro found the protein of the predicted 18r = 18 kDa. Recently, we have generated mutants of this protein by site- specific mutagenesis to confirm its role in this transactivation and HAVE replication.This study is important to better understand the role of HHV- 6 in HIV pathogenesis. The project will also provide knowledge and tools for development of novel antiviral therapies and safe viral vectors for gene therapy of AIDS.