GENETIC INTERACTION OF HUMAN HERPESVIRUS 6 WITH HIV 1

人类疱疹病毒 6 与 HIV 1 的基因相互作用

• 批准号: 2568936
• 负责人: A RAZZAQUE
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2568936
• 关键词: AIDS; animal tissue; chemical binding; cytomegalovirus; gel mobility shift assay; gene interaction; genetic promoter element; human herpesvirus 6; human immunodeficiency virus 1; human tissue; molecular site; site directed mutagenesis; transcription factor; transfection; transforming virus; virus genetics; virus replication; virus virus interaction

Studies on interactions of human herpesvirus-6 (HHV-6) or human cytomegalovirus (HCMV) with human immunodeficiency virus type 1 (HIV-1) are of interest in view of the suggested relationship between HHV-6 or CMV infections and the pathogenesis of AIDS. HHV-6 and HIV-1 can productively infect CD4+T cells and coinfection accelerates cytopathic effects. Studies were conducted to understand how these interactions occur at the molecular level. We tested HHV-6 genes that may affect HIV-1 replication. By DNA transfection into human T-cells and monkey kidney cells, we have shown that an HHV-6 transforming gene segment, ZVH14, previously identified in this laboratory, can transactivate the HIV-1 LTR. This transactivation is mediated through Sp1 binding sites in the HIV-1 long terminal repeat. A 115-amino acid open reading frame, B115, in the ZVH14 DNA showed a similar transactivation capacity as ZVH14. In vitro expression studies found the B115 gene product of the predicted Mr = 18 kDa. Mutants of this protein were generated by site-specific mutagenesis to confirm its role in transactivation. A specific mutant having a deletion of 5 nucleotides from the 5' - end of the ORF showed significant reduction in transactivation of the HIV-1 LTR. This observation supports our finding that B115 encodes a transactivator protein. In vitro expressed protein or the extracts from B115 transfected Cos-7 (SV40 transformed monkey kidney cells) cells did not show any binding to the HIV-1 LTR in gel shift assays. In a follow-up study, we have shown that B115 can also transactivate the CMV immediate early promoter in human fibroblasts and monkey kidney cells. Sp1 binding sites seem to mediate this transactivation as we have reported with the HIV-1 LTR activation. These data suggest that HHV-6 transforming gene segment, ZVH14, encodes a transactivator protein which can activate transacription from heterologous promoters and Sp1 binding sites are essential for this transactivation. This study is important for understanding the role of HHV-6 in the pathogenesis of AIDS and CMV retinitis. The project will also provide knowledge and tools for development of novel antiviral therapies and safe viral and plasmid vectors for gene therapy of AIDS.

人类疱疹病毒6型与人的相互作用研究 巨细胞病毒（HCMV）伴人类免疫缺陷病毒1型（HIV-1） 考虑到HHV-6或 CMV感染与艾滋病的发病机制。 HHV-6和HIV-1可以 有效感染CD 4 +T细胞，共感染加速细胞病变 方面的影响. 进行研究以了解这些相互作用 发生在分子水平上。 我们检测了HHV-6基因， HIV-1复制。 通过DNA转染人T细胞和猴T细胞 肾细胞，我们已经表明，HHV-6转化基因片段， ZVH 14，以前在这个实验室发现，可以反式激活 HIV-1 LTR。 这种反式激活通过Sp1结合位点介导 在HIV-1长末端重复序列中 115个氨基酸的开放阅读框， B115在ZVH 14 DNA中显示出与B115类似的反式激活能力， ZVH 14。 体外表达研究发现B115基因产物 预测Mr = 18 kDa。 该蛋白的突变体通过以下方法产生： 位点特异性诱变以确认其在反式激活中的作用。 一 具有从5' -末端缺失5个核苷酸的特异性突变体， ORF显示HIV-1 LTR的反式激活显著降低。 这一观察结果支持了我们的发现，即B115编码一个反式激活因子 蛋白 体外表达的蛋白或B115提取物 转染的Cos-7（SV 40转化的猴肾细胞）细胞不表达 在凝胶迁移试验中显示与HIV-1 LTR的任何结合。 新闻和跟踪 研究表明，B115也可以反式激活CMV立即 人成纤维细胞和猴肾细胞中的早期启动子。 Sp1结合 位点似乎介导了这种反式激活，正如我们已经报道的， HIV-1 LTR激活。 提示HHV-6转化基因 片段ZVH 14编码一种反式激活蛋白， 从异源启动子和Sp1结合位点的转录是 对这种反式激活至关重要。 本研究对于 了解HHV-6在AIDS和CMV发病机制中的作用 视网膜炎 该项目还将提供知识和工具， 新型抗病毒疗法和安全病毒和质粒的开发 用于艾滋病基因治疗的载体。