The acidic activation domains of the baculovirus transactivators IE1 and IE0 are functional for transcriptional activation in both insect and mammalian cells
The acidic activation domains of the baculovirus transactivators IE1 and IE0 are functional for transcriptional activation in both insect and mammalian cells
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DOI：
10.1099/vir.0.19747-0
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发表时间：
2004-03-01
影响因子：
3.8
通讯作者：
Theilmann, DA
中科院分区：
文献类型：
--
作者：
Dai, XJ;Willis, LG;Theilmann, DA
杆状病毒反式激活因子IE 1和IE 0的酸性激活结构域（AAD）对于转录反式激活至关重要。为了比较苜蓿银纹夜蛾核型多角体病毒（Autographa californica multiple nucleopolyhedrovirus，AcMNPV）和拟杉毒蛾核型多角体病毒（Orgyia pseudotsugata MNPV，OpMNPV）的IE 1和IE 0 AAD的相对转录激活潜力，我们构建了两个基于蜕皮激素受体（EcR）的诱导表达系统，并在两个昆虫细胞系（Ld 652 Y和Sf 9）和两个哺乳动物细胞系（NIH-3 T3和CHO）中分析了6种杆状病毒AAD。对于昆虫细胞表达，将AAD融合至云杉芽虫Choristoneura fumiferana EcR的C、D、E和F结构域。对于哺乳动物细胞表达，将AAD与哺乳动物小家鼠类维生素A X受体的E和F结构域融合。在Ld 652 Y和Sf 9细胞中，含有AcMNPV AAD的嵌合蛋白比含有OpMNPV AAD的嵌合蛋白更高水平地激活基因表达。在NIH-3 T3细胞中，含有AcMNPV IE 1和IE 0 AAD的嵌合体始终激活基因表达，使其水平高于原型哺乳动物疱疹病毒VP 16 AAD。相比之下，OpMNPV AAD相对于VP 16 AAD仅激活表达5-15%。在CHO细胞中，AcMNPV和OpMNPV AAD均表现出相对于VP 16 AAD的中间反式激活水平。这些结果表明，杆状病毒AAD在哺乳动物细胞中具有转录激活功能，并且AcMNPV AAD在昆虫和哺乳动物细胞中通常似乎比OpMNPV AAD更有效。
The acidic activation domains (AADs) of the baculovirus transactivators IE1 and IE0 are essential for transcriptional transactivation. To compare the relative transcriptional activation potentials of IE1 and IE0 AADs of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) and Orgyia pseudotsugata MNPV (OpMNPV), we constructed two ecdysone receptor (EcR)-based inducible expression systems to analyse six baculovirus AADs in two insect cell lines (Ld652Y and Sf9) and two mammalian cell lines (NIH-3T3 and CHO). For insect cell expression, the AADs were fused to the C, D, E and F domains of the spruce budworm Choristoneura fumiferana EcR. For mammalian cell expression the AADs were fused to the E and F domains of mammalian Mus musculus retinoid X receptor. In Ld652Y and Sf9 cells, chimeric proteins containing the AcMNPV AADs activated gene expression to higher levels than those containing the OpMNPV AADs. In NIH-3T3 cells, chimeras containing AcMNPV IE1 and IE0 AADs consistently activated gene expression to higher levels than the archetypal mammalian herpesvirus VP16 AAD. In contrast, OpMNPV AADs only activated expression by 5-15% relative to the VP16 AAD. In CHO cells, both AcMNPV and OpMNPV AADs exhibited intermediate transactivation levels relative to VP16 AAD. These results show that the baculovirus AADs are functional for transcriptional activation in mammalian cells and that AcMNPV AADs generally appear to be more potent than OpMNPV AADs in both insect and mammalian cells.