De novo RNA synthesis and homology modeling of the classical swine fever virus RNA polymerase

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	De novo RNA synthesis and homology modeling of the classical swine fever virus RNA polymerase
	Zhang, PW ; Xie, J ; Yi, GH ; Zhang, CY ; Zhou, R
刊名	VIRUS RESEARCH
	2005-09-01
卷号	112 期号:1-2 页码:9-23
关键词	de novo RNA synthesis homology modeling classical swine fever RNA polymerase
ISSN号	0168-1702
通讯作者	Zhang, CY, Guangzhou Children Hosp, Cent Lab, Guangzhou, Guangdong, Peoples R China
中文摘要	Classical swine fever virus (CSFV) non-structural protein 5B (NS5B) encodes an RNA-dependent RNA polymerase (RdRp), a key enzyme which initiates RNA replication by a de novo mechanism without a primer and is a potential target for anti-virus therapy. We expressed the NS5B protein in Escherichia coli. The rGTP can stimulate de novo initiation of RNA synthesis and mutation of the GDD motif to Gly-Asp-Asp (GAA) abolishes the RNA synthesis. To better understand the mechanism of viral RNA synthesis in CSFV, a three-dimensional model was built by homology modeling based on the alignment with several virus RdRps. The model contains 605 residues folded in the characteristic fingers, palm and thumb domains. The fingers domain contains an N-terminal region that plays an important role in conformational change. We propose that the experimentally observed promotion of polymerase efficiency by rGTP is probably due to the conformational changes of the polymerase caused by binding the rGTP. Mutation of the GDD to GAA interferes with the interaction between the residues at the polymerase active site and metal ions, and thus renders the polymerase inactive. (c) 2005 Elsevier B.V. All rights reserved.
英文摘要	Classical swine fever virus (CSFV) non-structural protein 5B (NS5B) encodes an RNA-dependent RNA polymerase (RdRp), a key enzyme which initiates RNA replication by a de novo mechanism without a primer and is a potential target for anti-virus therapy. We expressed the NS5B protein in Escherichia coli. The rGTP can stimulate de novo initiation of RNA synthesis and mutation of the GDD motif to Gly-Asp-Asp (GAA) abolishes the RNA synthesis. To better understand the mechanism of viral RNA synthesis in CSFV, a three-dimensional model was built by homology modeling based on the alignment with several virus RdRps. The model contains 605 residues folded in the characteristic fingers, palm and thumb domains. The fingers domain contains an N-terminal region that plays an important role in conformational change. We propose that the experimentally observed promotion of polymerase efficiency by rGTP is probably due to the conformational changes of the polymerase caused by binding the rGTP. Mutation of the GDD to GAA interferes with the interaction between the residues at the polymerase active site and metal ions, and thus renders the polymerase inactive. (c) 2005 Elsevier B.V. All rights reserved.
学科主题	Virology
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Virology
研究领域[WOS]	Virology
关键词[WOS]	HEPATITIS-C VIRUS ; VIRAL DIARRHEA VIRUS ; SECONDARY STRUCTURE PREDICTION ; MULTIPLE SEQUENCE ALIGNMENT ; REVERSE-TRANSCRIPTASE ; CRYSTAL-STRUCTURE ; ESCHERICHIA-COLI ; STRAND RNA ; ANGSTROM RESOLUTION ; ENZYMATIC-ACTIVITY
收录类别	SCI
语种	英语
WOS记录号	WOS:000231004300002
公开日期	2010-10-13
内容类型	期刊论文
源URL	[http://ir.ihb.ac.cn/handle/152342/9182]
专题	水生生物研究所_中科院水生所知识产出（2009年前）_期刊论文
作者单位	1.Guangzhou Children Hosp, Cent Lab, Guangzhou, Guangdong, Peoples R China 2.Chinese Acad Sci, Inst Hydrobiol, Wuhan, Hubei, Peoples R China
推荐引用方式 GB/T 7714	Zhang, PW,Xie, J,Yi, GH,et al. De novo RNA synthesis and homology modeling of the classical swine fever virus RNA polymerase[J]. VIRUS RESEARCH,2005,112(1-2):9-23.
APA	Zhang, PW,Xie, J,Yi, GH,Zhang, CY,&Zhou, R.(2005).De novo RNA synthesis and homology modeling of the classical swine fever virus RNA polymerase.VIRUS RESEARCH,112(1-2),9-23.
MLA	Zhang, PW,et al."De novo RNA synthesis and homology modeling of the classical swine fever virus RNA polymerase".VIRUS RESEARCH 112.1-2(2005):9-23.