Novel induction by herpes simplex virus of hybrid interferon gene transcripts driven by the strong cytomegalovirus IE94 promoter.
Novel induction by herpes simplex virus of hybrid interferon gene transcripts driven by the strong cytomegalovirus IE94 promoter.
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单纯疱疹病毒由强巨细胞病毒 IE94 启动子驱动的杂合干扰素基因转录物的新诱导。
DOI：
10.1128/jvi.61.3.819-828.1987
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发表时间：
1987
影响因子：
5.4
通讯作者：
Hayward,GS
中科院分区：
文献类型：
--
作者：
Mosca,JD;Jeang,KT;Pitha,PM;Hayward,GS
我们已经构建了稳定的DNA转染的LTK+细胞系含有两个不同的共选择的杂交干扰素（IFN）基因驱动的通常强大的和组成型启动子从猴巨细胞病毒的立即早期94 K蛋白（IE94）基因。令人惊讶的是，与杂合IE94-氯霉素乙酰转移酶基因构建体不同，两种IE94-IFN基因（一种具有和一种不具有复杂的剪接内含子区）在小鼠细胞系中产生相对低的生物活性人IFN的基础滴度。然而，IFN的表达可以刺激高达120倍的重叠感染单纯疱疹病毒（HSV），虽然不是与巨细胞病毒。为了研究这种意想不到的HSV诱导过程的机制，我们测量了在各种感染方案下产生的IE94-IFN mRNA和IFN蛋白的水平。与类似的先前表征的细胞系含有杂合IFN基因的HSV IE或延迟早期（DE）启动子的控制下，IFN表达的激活首先发生在中间时间。这两种IE94-IFN细胞系也产生了一种不寻常的模式，对HSV IE调节缺陷突变体tsK和tsB 7感染的反应：在没有功能性HSV IE 175（或ICP 4）基因产物的情况下，IFN合成受到刺激，但在没有病毒衣壳脱壳的情况下，IFN合成没有受到刺激。放线菌酮处理（无病毒感染）也给出了一个快速的30倍的稳定状态水平的正确启动的mRNA从两种类型的IE94-IFN杂合基因的增加，但没有影响的细胞含有IE 175-IFN结构。因此，我们得出结论，IE94-IFN构建体的使用鉴定了一种新的HSV调节应答，其需要HSV的先前未识别的功能，并且不涉及已知分别刺激HSV IE和DE基因转录的IE 175或前IE“病毒体因子”反式激活因子。
We have constructed stable DNA-transfected LTK+ cell lines containing two different coselected hybrid interferon (IFN) genes driven by the usually strong and constitutive promoter from the immediate-early 94K protein (IE94) gene of simian cytomegalovirus. Surprisingly, and unlike hybrid IE94-chloramphenicol acetyltransferase gene constructs, both of the IE94-IFN genes (one with and one without the complex spliced intron region) produced relatively low basal titers of biologically active human IFN in the mouse cell lines. However, IFN expression could be stimulated up to 120-fold by superinfection with herpes simplex virus (HSV), although not with cytomegalovirus. To examine the mechanism of this unexpected HSV induction process, we measured the levels of both IE94-IFN mRNA and IFN protein produced under various infection protocols. Compared with similar previously characterized cell lines containing hybrid IFN genes under the control of HSV IE or delayed-early (DE) promoters, activation of IFN expression first occurred at an intermediate time. Both IE94-IFN cell lines also produced an unusual pattern of response to infection with the HSV IE regulation-deficient mutants tsK and tsB7: stimulation of IFN synthesis occurred in the absence of a functional HSV IE175 (or ICP4) gene product, but did not occur in the absence of uncoating of virus capsids. Cycloheximide treatment (without virus infection) also gave a rapid 30-fold increase in steady-state levels of correctly initiated mRNA from both types of IE94-IFN hybrid genes, but had no effect on cells containing the IE175-IFN construct. Therefore, we conclude that the use of the IE94-IFN constructs identifies a novel HSV regulatory response that requires a previously unrecognized function of HSV and does not involve either IE175 or the pre-IE "virion factor" trans-activators that are known to stimulate transcription of HSV IE and DE genes, respectively.