trans-activation of the HIV promoter by a cDNA and its genomic clones of human herpesvirus-6.
trans-activation of the HIV promoter by a cDNA and its genomic clones of human herpesvirus-6.
复制标题
人类疱疹病毒 6 的 cDNA 及其基因组克隆对 HIV 启动子的反式激活。
作者：
Zhou,Y;Chang,CK;Qian,G;Chandran,B;Wood,C
人类疱疹病毒6型(HHV-6)是一种嗜淋巴的疱疹病毒，在体外能以HIV-1的形式感染人类CD4+T细胞。HIV-1和HHV-6共同感染T细胞可导致HIV-1启动子的激活和细胞病变的加速。编码41 kDa核蛋白的HHV-6(GS)基因克隆pCD41已被鉴定和鉴定(Chang和Balachandran，J.Viro.65,2884-2894和7085,1991)。序列分析表明，该蛋白与人类巨细胞病毒UL44基因编码的ICP36家族早晚期磷蛋白具有显著的血液学特征。以此为探针，克隆了编码HHV-6(GS)P41的3.8kbEcoRI基因组片段，命名为pGD41。当pCD41和pGD41克隆与HIV LTRCAT共转染入CV-1细胞后，均能激活HIV LTR。序列分析表明，pCD41有两个潜在的开放阅读框架(ORF)，A和B，它们与基因组克隆pGD41中发现的ORF同源。PCD41克隆的缺失构建表明ORF-A在HIV LTR激活中起关键作用。PCD41 ORF-A的缺失分析和启动子构建体的使用进一步定位了pCD41序列中的一个内部功能启动子，该启动子可以指导TRANS激活蛋白的合成。通过使用HIV LTR缺失突变体，发现NF-κB结合位点对pCD41反式激活的反应是关键的。
Human herpesvirus 6 (HHV-6) is a lymphotropic herpesvirus, andin vitro, it can productively infect human CD4+T cells as HIV-1. Co-infection of T cells by HIV-1 and HHV-6 can lead to both activation of the HIV-1 promoter and acceleration of the cytopathic effects. An HHV-6 (GS) cDNA clone, pCD41, encoding for a 41-kDa nuclear protein was identified and characterized previously (Chang and Balachandran,J. Virol.65, 2884-2894 and 7085, 1991). Sequence analyses show that this protein has significant hemology with the human cytomegalovirus UL44 gene coding for the ICP36 family of early-late-class phosphoprotein. Using this cDNA as the probe, a 3.8-kbEcoRI genomic fragment encoding the HHV-6(GS)P41 was cloned and designated as pGD41. When cotransfected with the HIV LTR CAT into CV-1 cells, both the pCD41 and pGD41 clonestrans-activated the HIV LTR. Sequence analyses of pCD41 indicate that there are two potential open reading frames (ORFs), A and B, which are homologous to the ORFs found in the genomic clone pGD41. Deletion constructs of the pCD41 clone demonstrated that ORF-A was critical for the HIV LTR activation. Deletion analyses of the pCD41 ORF-A and the use of promoter constructs further mapped an internal functional promoter within the pCD41 sequence that can direct the synthesis of thetrans-activating protein. By using HIV LTR deletion mutants, the NF-κB binding sites were found to be critical for response to the pCD41trans-activation.