腺病毒DNA聚合酶的核运输是通过与前终蛋白的相互作用来促进的
1Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City 66103.
Cell
|December 23, 1988
概括
腺病毒前终蛋白 (pTP) 促进DNA聚合酶 (AdPol) 的核运输. 这种互动对AdPol来说至关重要.
科学领域:
- 分子生物学分子生物学
- 病毒学 病毒学
- 细胞生物学 细胞生物学
背景情况:
- 腺病毒mRNA编码来自E2B转录单元的前终蛋白 (pTP) 和DNA聚合酶 (AdPol).
- 这些蛋白质含有主要的开放式读取框架 (m-ORFs) 与拼接的外子.
- 了解蛋白质定位和功能是病毒复制的关键.
研究的目的:
- 调查腺病毒pTP和AdPol的功能活动和亚细胞局部.
- 为了识别pTP和AdPol的核定位信号 (NLS).
- 确定pTP-AdPol相互作用在AdPol核进口中的作用.
主要方法:
- 在子脏细胞 (CV1) 中,pTP和AdPol变体 (pTP1,pTP2,AdPol1,AdPol2) 的过渡表达.
- 功能性检测:在体外复制启动.
- 使用显微镜进行亚细胞局部化研究.
- 共同表达实验以评估蛋白质相互作用.
主要成果:
- 只有从链接器ATG启动的pTP2和AdPol2,在复制启动中是活跃的.
- pTP (两种变体) 包含一个NLS (RLPV(R) 6VP) 并是核.
- AdPol1是细胞质的,而AdPol2位于细胞质和核中,这表明它的NLS位于前139个氨基酸中.
- pTP和AdPol的共同表达显著增加了AdPol的核分布.
- pTP促进了AdPol的核运输,独立于AdPol的NLS.
结论:
- 腺病毒pTP具有功能性的核定位信号.
- 亚地诺病毒AdPol的核进口因与pTP的相互作用而显著增强.
- 通过pTP介导的AdPol的核运输是腺病毒复制的关键步骤.
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