腺病毒VA RNAl与蛋白激酶DAI的相互作用:结合和功能不等价
K H Mellits1, M Kostura, M B Mathews
1Cold Spring Harbor Laboratory, New York 11724.
Cell
|June 1, 1990
概括
腺病毒VARNAI阻止蛋白质激酶DAI的激活,维持蛋白质合成. 它的结构至关重要,不同的域介于DAI的结合和抑制.
科学领域:
- 分子病毒学分子病毒学
- 蛋白质激酶调节 蛋白质激酶调节
- RNA结构-功能关系
背景情况:
- 腺病毒蛋白质合成依赖VA RNAI来抵消宿主防御.
- 双链RNA (dsRNA) 依赖的蛋白质激酶DAI抑制了蛋白质合成.
- 众所周知,VA RNAI可以抑制DAI的激活,但确切的机制尚不清楚.
研究的目的:
- 研究VARNAI结构及其结合和抑制DAI的能力之间的关系.
- 为了确定VA RNAI的特定结构元素,负责DAI相互作用和抑制.
主要方法:
- 在体外结合测定使用纯化的DAI和野生类型/突变VARNA.
- 紫外线交叉链接以确认VA RNAI-DAI相互作用.
- 对VARNAs的二次结构分析.
- 功能性测试以评估DAI抑制在体内.
主要成果:
- VA RNAI与纯化的DAI结合,这种相互作用可以通过UV交叉链接来证实.
- 在体外,VA RNAI与DAI的有效结合并不总是与体内功能相关.
- 对VARNAI与DAI结合而言,一个顶端的茎环结构很重要.
- VA RNAI的中心域对于抑制DAI激活至关重要.
结论:
- VA RNAI利用不同的结构域与DAI结合并抑制其活动.
- VA RNAI的双重干可能与DAI上的dsRNA结合部位相互作用.
- VA RNAI通过将其中心域定位在酶活性部位附近来抑制DAI的激活.
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