Regulation of cytomegalovirus gene expression: alpha and beta promoters are trans activated by viral functions in permissive human fibroblasts.
Regulation of cytomegalovirus gene expression: alpha and beta promoters are trans activated by viral functions in permissive human fibroblasts.
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巨细胞病毒基因表达的调节：α和β启动子在允许的人成纤维细胞中被病毒功能反式激活。
DOI：
10.1128/jvi.56.1.135-143.1985
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发表时间：
1985
影响因子：
5.4
通讯作者：
Mocarski,ES
中科院分区：
文献类型：
--
作者：
Spaete,RR;Mocarski,ES
我们将从巨细胞病毒（CMV）基因组中提取的即时（α）和延迟（β）早期启动子调控序列融合到大肠杆菌lacZ （β -半乳糖苷酶）中作为指示基因来研究这些启动子的调控表达。用携带β -半乳糖苷酶融合物的质粒转染人成纤维细胞并随后感染巨细胞病毒后，我们已经证明病毒反式作用功能以暂时真实的方式上调了这些基因的表达。即使在新生蛋白合成被阻断和使用紫外线灭活病毒时，α启动子也被激活，这表明，对于1型单纯疱疹病毒（HSV-1），一种病毒粒子结构蛋白负责其上调。我们发现HSV-1和巨细胞病毒都能够反式激活巨细胞病毒α启动子。启动子可被巨细胞病毒激活，但对1型单纯疱疹病毒感染完全无反应。α启动子和β启动子在瞬时表达系统中的合成符合它们在病毒复制过程中的自然调控。我们描述的β -半乳糖苷酶融合体为研究顺式和反式作用的病毒调节功能提供了一个最灵敏的指标系统。
We have fused immediate (alpha) and delayed (beta) early promoter-regulatory sequences taken from the cytomegalovirus (CMV) genome to Escherichia coli lacZ (beta-galactosidase) as an indicator gene to study regulated expression of these promoters. After transfection of human fibroblast cells with plasmid constructs carrying beta-galactosidase fusions, and subsequent infection with CMV, we have demonstrated that viral trans-acting functions up-regulate the expression of these genes in a temporally authentic manner. The alpha promoter is activated even when de novo protein synthesis is blocked and when UV-inactivated virus is used, suggesting that, as for herpes simplex virus type 1 (HSV-1), a virion structural protein is responsible for its up-regulation. We have found that HSV-1, as well as CMV, is capable of trans activating the CMV alpha promoter. The beta promoter is activated by CMV but is completely unresponsive to HSV-1 infection. The temporal synthesis of the alpha and beta promoters in the transient expression system conforms with their natural regulation during viral replication. The beta-galactosidase fusions we describe provide a most exquisitely sensitive indicator system for the study of cis- and trans-acting viral regulatory functions.