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首页> 外文期刊>Molecular and Cellular Biology >trans activation of an Epstein-Barr viral transcriptional enhancer by the Epstein-Barr viral nuclear antigen 1.
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trans activation of an Epstein-Barr viral transcriptional enhancer by the Epstein-Barr viral nuclear antigen 1.

机译：Epstein-Barr病毒核抗原1.反式激活Epstein-Barr病毒转录增强子。

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Two regions of the Epstein-Barr virus (EBV) genome together make up an element, oriP, which acts in cis to support plasmid replication in cells that express the EBV nuclear antigen 1 (EBNA-1). The two components of oriP are a region containing a 65-base-pair (bp) dyad symmetry and a region containing 20 copies of a 30-bp direct repeat. Here we show that the 30-bp family of repeats of oriP can function as a transcriptional enhancer that is activated in trans by the EBNA-1 gene product. In either EBV-genome-positive cells or in cells that express EBNA-1, the 30-bp family of repeats, when positioned in either orientation upstream or downstream, enhances expression of the chloramphenicol acetyltransferase (CAT) gene expressed from either the simian virus 40 early promoter or the herpes simplex virus type 1 thymidine kinase promoter. The extent of transcriptional enhancement varies with the promoter and cell type. This enhanced CAT expression reflects an increased level of CAT mRNA and does not result from amplification of the plasmids expressing CAT. In addition, plasmids carrying the gene for resistance to hygromycin B and the 30-bp family of repeats yielded 10 to 100 times more hygromycin B-resistant colonies than the vector lacking the 30-bp family of repeats in both EBV-genome-positive cells and cells that express EBNA-1. EBNA-1 is known to bind to sequences within the 30-bp family of repeats (D. R. Rawlins, G. Milman, S. D. Hayward, and G. S. Hayward, Cell 42:859-868, 1985), and these trans- and cis-acting elements together have at least two functional roles: (i) they are required for DNA replication dependent upon oriP, and (ii) they can enhance expression of genes linked to the 30-bp family of repeats of oriP.
机译：爱泼斯坦-巴尔病毒（EBV）基因组的两个区域共同构成一个元素oriP，它以顺式作用支持在表达EBV核抗原1（EBNA-1）的细胞中的质粒复制。 oriP的两个组成部分是包含65个碱基对（bp）dyad对称性的区域和包含20个30 bp直接重复的拷贝的区域。在这里，我们显示oriP的30 bp重复序列家族可以充当转录增强子，该转录增强子被EBNA-1基因产物反式激活。在EBV基因组阳性细胞或表达EBNA-1的细胞中，当重复序列位于上游或下游时，其30 bp重复序列家族可增强猿猴病毒表达的氯霉素乙酰转移酶（CAT）基因的表达。 40个早期启动子或1型单纯疱疹病毒胸苷激酶启动子。转录增强的程度随启动子和细胞类型而变化。这种增强的CAT表达反映了CAT mRNA水平的增加，而不是由表达CAT的质粒的扩增引起的。此外，在两个EBV基因组阳性细胞中，携带抗潮霉素B和30 bp重复序列的基因的质粒产生的抗潮霉素B的菌落比缺乏30 bp重复序列的载体高10到100倍和表达EBNA-1的细胞。已知EBNA-1与30 bp重复序列家族的序列结合（DR Rawlins，G.Milman，SD Hayward和GS Hayward，Cell 42：859-868，1985），并且这些反式和顺式作用元件一起具有至少两个功能作用：（i）它们是依赖oriP的DNA复制所必需的;（ii）它们可以增强与oriP的30-bp重复序列相关的基因的表达。

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