Interaction between SARS-CoV helicase and a multifunctional cellular protein (Ddx5) revealed by yeast and mammalian cell two-hybrid systems.
Interaction between SARS-CoV helicase and a multifunctional cellular protein (Ddx5) revealed by yeast and mammalian cell two-hybrid systems.
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DOI：
10.1007/s00705-009-0323-y
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发表时间：
2009
影响因子：
2.7
通讯作者：
Wen YM
中科院分区：
文献类型：
--
作者：
Chen JY;Chen WN;Poon KM;Zheng BJ;Lin X;Wang YX;Wen YM
为了揭示SARS冠状病毒复制过程中可能涉及的细胞因子，以SARS冠状病毒解旋酶（Hel，nsp 13）为模板，利用酵母双杂交系统筛选大鼠肺上皮细胞cDNA文库。使用哺乳动物细胞杂交系统和人A549细胞系中的免疫共沉淀进一步测试了阳性相互作用蛋白，该细胞系已被证明支持SARS冠状病毒复制。在酵母双杂交检测的7个阳性克隆中，只有Ddx 5（Asp-Glu-Ala-Asp box polypeptide 5）蛋白与SARS-CoV解旋酶具有特异性相互作用。当用小干扰RNA（siRNA）敲低DdX 5的表达时，SARS冠状病毒在胎猴肾（FRhK-4）细胞中的复制被显著抑制。由于Ddx 5是一种多功能蛋白，在转录调控中起着重要作用，它与SARS冠状病毒解旋酶的相互作用为研究SARS冠状病毒感染中病毒-宿主细胞相互作用提供了有趣的线索。本文的在线版本（doi：10.1007/s 00705 -009-0323-y）包含补充材料，可供授权用户使用。
To reveal the putative cellular factors involved in SARS coronavirus replication, the helicase (Hel, nsp13) of SARS coronavirus was used to screen the cDNA library of rat pulmonary epithelial cells using the yeast two-hybrid system. Positively interacting proteins were further tested using a mammalian cell hybrid system and co-immunoprecipitation in the human A549 cell line, which has been shown to support SARS coronavirus replication. Out of the seven positive clones observed by yeast two-hybrid assay, only the Ddx5 (Asp-Glu-Ala-Asp box polypeptide 5) protein showed specific interaction with SARS-CoV helicase. When expression of DdX5 was knocked down by small interfering RNA (siRNA), SARS coronavirus replication was significantly inhibited in fetal rhesus kidney (FRhK-4) cells. Since Ddx5 is a multifunctional protein that plays important roles in transcriptional regulation, its interaction with SARS coronavirus helicase provides interesting clues for studying virus–host cell interactions in SARS-CoV infections. The online version of this article (doi:10.1007/s00705-009-0323-y) contains supplementary material, which is available to authorized users.
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影响因子：
8
作者：
Rossow, KL;Janknecht, R
通讯作者：
Janknecht, R
影响因子：
5.4
作者：
CRAWFORD, L;LEPPARD, K;HARLOW, E
通讯作者：
HARLOW, E
影响因子：
5.4
作者：
van Dinten, LC;van Tol, H;Snijder, EJ
通讯作者：
Snijder, EJ
影响因子：
82.9
作者：
Crute, JJ;Grygon, CA;Cordingley, MG
通讯作者：
Cordingley, MG
DOI：
10.1074/jbc.c300328200
发表时间：
2003-10-10
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Tanner JA;Watt RM;Chai YB;Lu LY;Lin MC;Peiris JS;Poon LL;Kung HF;Huang JD
通讯作者：
Huang JD