CHARACTERIZATION OF AN R-BINDING SITE MEDIATING THE R-INDUCED ACTIVATION OF THE EPSTEIN-BARR-VIRUS BMLF1 PROMOTER
CHARACTERIZATION OF AN R-BINDING SITE MEDIATING THE R-INDUCED ACTIVATION OF THE EPSTEIN-BARR-VIRUS BMLF1 PROMOTER
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DOI：
10.1128/jvi.66.1.46-52.1992
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发表时间：
1992-01-01
影响因子：
5.4
通讯作者：
SERGEANT, A
中科院分区：
文献类型：
--
作者：
GRUFFAT, H;DURAN, N;SERGEANT, A
在潜伏感染Epstein-Barr病毒的细胞中，从潜伏到生产感染的转变与两种Epstein-Barr病毒转录因子EB1和R的表达有关。R是一种增强因子，并且在BMLF1启动子中发现了R响应元件（RRE）。在这项研究中，我们使用双向缺失诱变技术将BMLF1 RRE （RRE- m）描述为一个44-bp的序列。我们还发现，从重组痘苗病毒中表达的R可以保护RRE-M免受DNA酶i的消化。利用迁移率转移试验和硫酸二甲酯干扰，我们表征了体外翻译的R和DNA之间的接触点。R在体外通过同时接触该位点内的两个序列与一个位点结合，这两个序列相距8bp: 5‘-catGTCCCtctatcatGGCGCagac-3’。该序列的定点突变在体外完全破坏了R的结合，使BMLF1启动子对R无反应。结果表明，R诱导的BMLF1增强子由一个单独的R结合位点组成，称为RRE-M。
In cells latently infected with Epstein-Barr virus, the switch from latency to productive infection is linked to the expression of two Epstein-Barr virus transcription factors called EB1 and R. R is an enhancer factor, and an R-responsive element (RRE) has been identified in the BMLF1 promoter. In this study, we have used bidirectional deletion mutagenesis to delineate the BMLF1 RRE (RRE-M) to a 44-bp sequence. We also show that R expressed from a recombinant vaccinia virus protects RRE-M against digestion by DNase I. Using mobility shift assays and dimethyl sulfate interferences, we have characterized the contact points between in vitro-translated R and the DNA. R binds in vitro to one site by simultaneously contacting two sequences within the site, which are separated by 8 bp: 5'-catGTCCCtctatcatGGCGCagac-3'. Site-directed mutagenesis of this sequence completely impaired the binding of R in vitro and rendered the BMLF1 promoter nonresponsive to R. The results suggest that the R-inducible BMLF1 enhancer is composed of a single R-binding site, called RRE-M.