REGULATION OF THE EPSTEIN-BARR-VIRUS DNA-POLYMERASE GENE
REGULATION OF THE EPSTEIN-BARR-VIRUS DNA-POLYMERASE GENE
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DOI：
10.1128/jvi.66.5.2837-2845.1992
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发表时间：
1992-05-01
影响因子：
5.4
通讯作者：
PAGANO, JS
中科院分区：
文献类型：
--
作者：
FURNARI, FB;ADAMS, MD;PAGANO, JS
编码EB病毒（EBV）DNA聚合酶的基因（pol），属于病毒基因的“早期”类别，在潜伏病毒感染激活后不久表达。 首先，分析了用12-O-十四酰基佛波醇-13-乙酸酯和丁酸钠处理以诱导裂解性复制和该基因表达的EBV产生细胞系B 95 -8的mRNA。 北方（RNA）分析揭示了仅在诱导的细胞中发现的3.7 kb的信息。 通过S1核酸酶和引物延伸分析对pol mRNA的5'定位表明，转录起始于开放阅读框上游126 bp的富含G + C的区域内的紧密聚集的位点。 诱导后，在另外两个EBV感染的细胞系P3 HR 1和Raji中鉴定出相同的起始区域。 第二，一个1.29 kb的基因组片段含有这个区域，当克隆的氯霉素乙酰转移酶报告基因的上游，表现出启动子活性与pEBV-RZ，基因组表达构建体，包括EBV立即早期反式激活蛋白，BZLF-1和BRLF-1基因共转染的淋巴细胞。 在上游1.29-kb序列内，140 bp和101 bp的两个区域似乎是启动子活性所需的。 这些结果表明，与迄今为止研究的大多数EBV基因不同，pol基因含有多个转录起始位点。 pol基因启动子的上游调控区不含典型的启动子元件，如TATA和CAAT盒，此外，不具有组成型活性，但需要两种或多种病毒蛋白的反式激活。
The gene (pol) encoding the Epstein-Barr virus (EBV) DNA polymerase of the "early" class of viral genes which are expressed shortly after activation of latent virus infection. First, mRNA from the EBV-producing cell line, B95-8, treated with 12-O-tetradecanoylphorbol-13-acetate and sodium butyrate to induce lytic replication and expression of this gene was analyzed. Northern (RNA) analysis revealed a message of 3.7 kb found only in induced cells. 5' mapping of pol mRNA by S1 nuclease and primer extension analyses indicates that transcription initiates at tightly clustered sites within a G + C-rich region 126 bp upstream of the open reading frame. The same initiation region was identified in two other EBV-infected cell lines, P3HR1 and Raji, after induction. Second, a 1.29-kb genomic fragment containing this region, when cloned upstream of the chloramphenicol acetyltransferase reporter gene, demonstrated promoter activity in lymphoid cells cotransfected with pEBV-RZ, a genomic expression construct that includes genes for the EBV immediate-early transactivator proteins, BZLF-1 and BRLF-1. Within the upstream 1.29-kb sequence, two regions of 140 bp and 101 bp appear to be needed for promoter activity. These results demonstrate that unlike most EBV genes studied thus far, the pol gene contains multiple transcriptional start sites. The upstream regulatory region of the promoter for the pol genes does not contain canonical promoter elements such as TATA and CAAT boxes and, furthermore, is not constitutively active but requires transactivation by two or more viral proteins.