Salamon Daniel, Takacs Maria, Schwarzmann Fritz, Wolf Hans, Minarovits Janos, Niller Hans Helmut
Microbiological Research Group, National Center for Epidemiology, Pihenö ut 1, H-1529 Budapest, Hungary.
Virus Genes. 2003 Aug;27(1):57-66. doi: 10.1023/a:1025124519068.
Latency protein LMP2A of Epstein-Barr virus (EBV) has been implicated in EBV related tumorigenesis. To understand the host cell dependent expression of the LMP2A gene, it is necessary to analyse the regulatory mechanisms of the LMP2A promoter (LMP2Ap). By transient transfection and in vitro binding analyses two CBF1 sites have previously been shown to be involved in the regulation of LMP2Ap. However, the promoter structure has not been examined at the nucleotide level in vivo. Therefore we undertook a comprehensive analysis of in vivo protein binding and of CpG-methylation patterns at LMP2Ap in a panel of B cell lines carrying latent EBV genomes. The presence of characteristic footprints on two CBF1 and further binding-sites, together with overall hypomethylation of CpG dinucleotides correlated well with promoter activity. In contrast, the absence of several genomic footprints, as well as the presence of patches of highly methylated CpG dinucleotides were characteristic of silent LMP2Aps.
爱泼斯坦-巴尔病毒（EBV）的潜伏蛋白LMP2A与EBV相关的肿瘤发生有关。为了解LMP2A基因的宿主细胞依赖性表达，有必要分析LMP2A启动子（LMP2Ap）的调控机制。通过瞬时转染和体外结合分析，先前已表明两个CBF1位点参与LMP2Ap的调控。然而，尚未在体内核苷酸水平上研究启动子结构。因此，我们对一组携带潜伏EBV基因组的B细胞系中LMP2Ap的体内蛋白质结合和CpG甲基化模式进行了全面分析。两个CBF1和其他结合位点上特征性足迹的存在，以及CpG二核苷酸的整体低甲基化与启动子活性密切相关。相反，几个基因组足迹的缺失以及高度甲基化的CpG二核苷酸区域的存在是沉默LMP2Ap的特征。
