牛泡沫病毒反式激活因子在内部启动子上应答元件的研究.docVIP

牛泡沫病毒反式激活因子在内部启动子上应答元件的研究 ???? 牛泡沫病毒反式激活因子在内部启动子上应答元件的研究 ???? 病毒学报 2000年第3期第16卷 论　著—人与动物病毒 ???? 作者：张莉　乔文涛　刘淑红　王金忠　陈启民　耿运琪 ???? 单位：南开大学生命科学学院，天津 300071 ???? 关键词：牛泡沫病毒；内部启动子（IP）；转录反式激活因子(Tas)；反式激活因子应答元件（TRE）；TATA盒 ???? 　　摘要：泡沫病毒的基因转录依赖至少两个不同的启动子：LTR调节病毒结构蛋白的表达，而内部启动子（IP）则起始调节蛋白mRNA的转录。牛泡沫病毒（BFV）在env与3′LTR之间有两个重叠的开放阅读框架orf-1和orf-2，分别编码BFV ORF-1、ORF-2等多种调节蛋白。这些蛋白中BFV ORF-1为转录激活因子，称为Tas。Tas对LTR及IP均有反式激活作用。BFV中第二类启动子IP的存在反映了泡沫病毒基因调控的复杂性。已从BFV3026中国毒株中克隆了基因组区段8 572～9 509，并通过测序和功能分析证明该区段包含了BFV IP。为了对IP进行更精确的定位，对其进行了进一步的缺失分析，将完整的IP定位在9 117～9 405之间。该启动子的TATA盒位于91 280～9 285，是IP必不可少的元件之一。杂合启动子和缺失分析表明，TATA盒上游160bp的区域内包含了IP的Tas应答元件（TREIP），其功能类似于增强子。Tas对IP具有强烈的激活作用，而C端缺失的ORF-1蛋白或ORF-2蛋白均不足以激活IP的表达。 ???? 　　中图分类号：S852.65+9.3; Q75 　　文献标识码：A ???? 　　文章编号：1000－8721（2000）03-0232-06 ???? Regulatory Elements That Mediate Transactivation by Tas in the Internal Promoter of Bovine Foamy Virus ???? ZHANG Li，QIAO Wen-tao，LIU Shu-hong， ???? 　　WANG Jin-zhon，CHEN Qi-min，GENG Yun-qi ???? 　　（College of Life Sciences, Nankai University, Tianjin 300071, China） ???? 　　Abstract: Foamy viruses are unusual among retroviruses in that they contain two distinct promoter elements. They are the long terminal repeat (LTR) promoter which regulates expression of the viral structural proteins, and a second internal promoter that locates towards the 3 end of the env gene and directs expression of the viral auxiliary proteins. One of these auxiliary proteins is a potent transcriptional transactivator, termed Tas in BFV. That is critical for foamy virus replication. Previously, we have cloned the intact BFV IP and demonstrated it to be highly responsive to the Tas protein. Here, we extended this earlier work by using deleted mutants and hybrid construct to localize the IP more precisely and to identify and characterize the Tas responsive element (TREIP) . Based on these data, we propose that genomic region 9117～9405nt contains all the sequences required for full Tas responsiveness. The importance of a TAT