GENETIC INTERACTION OF HUMAN HERPESVIRUS-6 WITH HIV-1

该项目的目标是识别和描述人类 疱疹病毒-6（HHV-6）基因，可激活人类免疫缺陷 病毒1型（HAVE）启动子，并确定如何相互作用 HHV-6被认为是这两种病毒之间的辅因子。 在HAVE感染和艾滋病的发展中，由于1）HHV-6 能有效地感染许多与HIV-6相同的细胞类型， 通过DNA转染到猴肾中， 细胞和人T细胞，我们已经表明，HHV-6基因片段ZVH 14 (8.7 kb），发现其可转化小鼠细胞和人类细胞， 角质形成细胞，可以增加该指标的表达 氯霉素乙酰转移酶zCAT）基因与HAVE启动子连接。 这种激活通过Sp1（细胞因子）结合位点介导 存在于HAVE启动子中，并且激活被显著降低， 寡聚体被设计为阻断Sp1结合位点。 用于有效 激活时，需要一个以上的Sp1位点。 NF-kB位点 HAVE启动子对HHV-6 ZVH 12介导的激活不是必需的。 我们对ZVH 14缺失锥的研究表明，一个小的开放的 称为B115的阅读框，可能编码115个氨基酸的蛋白质， 主要参与了这种转录激活功能。 初步 体外表达研究发现预测的18 r = 18的蛋白质 kDa。 最近，我们已经产生了这种蛋白质的突变体的网站- 特异性诱变以确认其在这种反式激活中的作用， 这项研究对于更好地理解HHV的作用很重要。 6.艾滋病发病机制。 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.