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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; Zhang, CY, Guangzhou Children Hosp, Cent Lab, Guangzhou, Guangdong, Peoples R China
2005-09-01
Source PublicationVIRUS RESEARCH
ISSN0168-1702
Volume112Issue:1-2Pages:9-23
AbstractClassical 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.; 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.
SubtypeArticle
KeywordDe Novo Rna Synthesis Homology Modeling Classical Swine Fever Rna Polymerase
DepartmentGuangzhou Children Hosp, Cent Lab, Guangzhou, Guangdong, Peoples R China; Chinese Acad Sci, Inst Hydrobiol, Wuhan, Hubei, Peoples R China
Subject AreaVirology
DOI10.1016/j.virusres.2005.03.003
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Indexed BySCI
Language英语
WOS Research AreaVirology
WOS SubjectVirology
WOS IDWOS:000231004300002
WOS KeywordHEPATITIS-C VIRUS ; VIRAL DIARRHEA VIRUS ; SECONDARY STRUCTURE PREDICTION ; MULTIPLE SEQUENCE ALIGNMENT ; REVERSE-TRANSCRIPTASE ; CRYSTAL-STRUCTURE ; ESCHERICHIA-COLI ; STRAND RNA ; ANGSTROM RESOLUTION ; ENZYMATIC-ACTIVITY
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Cited Times:9[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/9182
Collection期刊论文
Corresponding AuthorZhang, CY, Guangzhou Children Hosp, Cent Lab, Guangzhou, Guangdong, Peoples R China
Affiliation1.Guangzhou Children Hosp, Cent Lab, Guangzhou, Guangdong, Peoples R China
2.Chinese Acad Sci, Inst Hydrobiol, Wuhan, Hubei, Peoples R China
Recommended Citation
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,&Zhang, CY, Guangzhou Children Hosp, Cent Lab, Guangzhou, Guangdong, Peoples R China.(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.
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