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酵母和哺乳动物细胞双杂交系统揭示的严重急性呼吸综合征冠状病毒解旋酶与多功能细胞蛋白（Ddx5）之间的相互作用

Interaction between SARS-CoV helicase and a multifunctional cellular protein (Ddx5) revealed by yeast and mammalian cell two-hybrid systems.

作者信息

Chen Jin-Yan, Chen Wan-Nan, Poon Kwok-Man Vincent, Zheng Bo-Jian, Lin Xu, Wang Yong-Xiang, Wen Yu-Mei

机构信息

Key Laboratory of Infection and Oncology, Research Center of Molecular Medicine, Fujian Medical University, 350004, Fuzhou, People's Republic of China.

出版信息

Arch Virol. 2009;154(3):507-12. doi: 10.1007/s00705-009-0323-y. Epub 2009 Feb 18.

DOI:10.1007/s00705-009-0323-y
PMID:19224332
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7087151/
Abstract

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.

摘要

为了揭示参与严重急性呼吸综合征冠状病毒（SARS-CoV）复制的假定细胞因子，利用酵母双杂交系统，以SARS-CoV解旋酶（Hel，nsp13）筛选大鼠肺上皮细胞的cDNA文库。利用哺乳动物细胞杂交系统和在已被证明支持SARS-CoV复制的人A549细胞系中进行的免疫共沉淀，对阳性相互作用蛋白进行进一步检测。在酵母双杂交试验观察到的7个阳性克隆中，只有Ddx5（天冬氨酸-谷氨酸-丙氨酸-天冬氨酸盒多肽5）蛋白与SARS-CoV解旋酶表现出特异性相互作用。当用小干扰RNA（siRNA）敲低DdX5的表达时，SARS-CoV在恒河猴胎儿肾（FRhK-4）细胞中的复制受到显著抑制。由于Ddx5是一种在转录调控中起重要作用的多功能蛋白，其与SARS-CoV解旋酶的相互作用为研究SARS-CoV感染中的病毒-宿主细胞相互作用提供了有趣的线索。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/490b/7087151/87a4919a2a06/705_2009_323_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/490b/7087151/aaff4c5f5c0a/705_2009_323_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/490b/7087151/2e5b6972a4ab/705_2009_323_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/490b/7087151/87a4919a2a06/705_2009_323_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/490b/7087151/aaff4c5f5c0a/705_2009_323_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/490b/7087151/2e5b6972a4ab/705_2009_323_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/490b/7087151/87a4919a2a06/705_2009_323_Fig3_HTML.jpg

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