Characterization of the acidic domain of the IE1 regulatory protein from Orgyia pseudotsugata multicapsid nucleopolyhedrovirus
Characterization of the acidic domain of the IE1 regulatory protein from Orgyia pseudotsugata multicapsid nucleopolyhedrovirus
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DOI：
10.1006/viro.1998.9415
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发表时间：
1998-12-05
期刊：
影响因子：
3.7
通讯作者：
Theilmann, DA
中科院分区：
文献类型：
--
作者：
Forsythe, IJ;Shippam, CE;Theilmann, DA
本研究提出了一个详细的分析酸性N-末端区域的Orgyia pseudotsugata多衣壳核多角体病毒（OpMNPV）的反式激活因子IE 1。IE 1的N-末端区域富含酸性氨基酸，并被假设为酸性激活结构域。去除含有IE 1酸性结构域的N-末端126个氨基酸导致反式激活活性完全丧失，表明该区域对于反式激活是必需的。OpMNPV酸性结构域被来自VP 16的原型酸性激活结构域和苜蓿银纹夜蛾多衣壳NPV（AcMNPV）IE 1的富酸区域替换。这些嵌合构建体在瞬时测定中完全能够反式激活。含有单纯疱疹病毒VP 16和AcMNPV IE 1酸性激活结构域的嵌合体OpMNPV IE 1始终反式激活报告基因至比OpMNPV IE 1酸性激活结构域更高的水平。当与IE 2共转染到舞毒蛾和草地贪夜蛾细胞中时，嵌合构建体的反式激活协同增强。构建了OpMNPV酸性激活结构域的N-至C-末端和C-至N-末端缺失，以确定OpMNPV IE 1酸性激活结构域内的功能结构域。至少有两个潜在的激活域被确定。在这些结构域中的每一个内，鉴定了氨基酸28-43和氨基酸113-124处的两个核心区，其与主要含有酸性和大体积疏水性氨基酸的VP 16和GAL 4的核心区相似。(C)北京：科学出版社.
This study presents a detailed analysis of the acidic N-terminal region of the Orgyia pseudotsugata multicapsid nucleopolyhedrovirus (OpMNPV) transactivator IE1. The N-terminal region of IE1 is rich in acidic amino acids and has been hypothesized to be an acidic activation domain. Removal of the N-terminal 126 amino acids containing the acidic domain of IE1 resulted in complete loss of transactivation activity, indicating that this region is essential for transactivation. The OpMNPV acidic domain was replaced with the archetype acidic activation domain from VP16 and the acid-rich region of Autographa californica multicapsid NPV (AcMNPV) IE1. These chimeric constructs were fully capable of transactivation in transient assays. The chimeric OpMNPV IE1s containing the herpes simplex virus VP16 and AcMNPV IE1 acidic activation domains consistently transactivated a reporter gene to higher levels than the OpMNPV IE1 acidic activation domain. Transactivation by the chimeric constructs is enhanced synergistically when cotransfected with IE2 into Lymantria dispar and Spodoptera frugiperda cells. Both N- to C-terminal and C- to N-terminal deletions of the OpMNPV acidic activation domain were constructed to define functional domains within the OpMNPV IE1 acidic activation domain. At least two potential activation domains were identified. Within each of these domains, two core regions at amino acids 28-43 and amino acids 113-124 were identified that were similar to core regions of VP16 and GAL4, which contain predominately acidic and bulky hydrophobic amino acids. (C) 1998 Academic Press.