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Characterization of an early gene encoding for dUTPase in Rana grylio virus
Zhao, Zhe; Ke, Fei; Gui, Jianfang; Zhang, Qiya; Zhang, QY, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
2007-02-01
Source PublicationVIRUS RESEARCH
ISSN0168-1702
Volume123Issue:2Pages:128-137
AbstractdUTPase (DUT) is a ubiquitous and important enzyme responsible for regulating levels of dUTP. Here, an iridovirus DUT was identified and characterized from Rana grylio virus (RGV) which is a pathogen agent in pig frog. The DUT encodes a protein of 164aa with a predicted molecular mass of 17.4 kDa, and its transcriptional initiation site was determined by 5'RACE to start from the nucleotide A at 15 nt upstream of the initiation codon ATG. Sequence comparisons and multiple alignments suggested that RGV DUT was quite similar to other identified DUTs that function as homotrimers. Phylogenetic analysis implied that DUT horizontal transfers might have occurred between the vertebrate hosts and iridoviruses. Furthermore, its temporal expression pattern during RGV infection course was characterized by RT-PCR and Western blot analysis. It begins to transcribe and translate as early as 4 h postinfection (p.i.), and remains detectable at 48 h p.i. DUT-EGFP fusion protein was observed in the cytoplasm of pEGFP-N3-Dut transfected EPC cells. Immunofluorescence also confirmed DUT cytoplasm localization in RGV-infected cells. Using drug inhibition analysis by a de novo protein synthesis inhibitor (cycloheximide) and a viral DNA replication inhibitor (cytosine arabinofuranoside), RGV DUT was classified as an early (E) viral gene during the in vitro infection. Moreover, RGV DUT overexpression was shown that there was no effect on RGV replication by viral replication kinetics assay. (c) 2006 Published by Elsevier B.V.; dUTPase (DUT) is a ubiquitous and important enzyme responsible for regulating levels of dUTP. Here, an iridovirus DUT was identified and characterized from Rana grylio virus (RGV) which is a pathogen agent in pig frog. The DUT encodes a protein of 164aa with a predicted molecular mass of 17.4 kDa, and its transcriptional initiation site was determined by 5'RACE to start from the nucleotide A at 15 nt upstream of the initiation codon ATG. Sequence comparisons and multiple alignments suggested that RGV DUT was quite similar to other identified DUTs that function as homotrimers. Phylogenetic analysis implied that DUT horizontal transfers might have occurred between the vertebrate hosts and iridoviruses. Furthermore, its temporal expression pattern during RGV infection course was characterized by RT-PCR and Western blot analysis. It begins to transcribe and translate as early as 4 h postinfection (p.i.), and remains detectable at 48 h p.i. DUT-EGFP fusion protein was observed in the cytoplasm of pEGFP-N3-Dut transfected EPC cells. Immunofluorescence also confirmed DUT cytoplasm localization in RGV-infected cells. Using drug inhibition analysis by a de novo protein synthesis inhibitor (cycloheximide) and a viral DNA replication inhibitor (cytosine arabinofuranoside), RGV DUT was classified as an early (E) viral gene during the in vitro infection. Moreover, RGV DUT overexpression was shown that there was no effect on RGV replication by viral replication kinetics assay. (c) 2006 Published by Elsevier B.V.
SubtypeArticle
KeywordRana Grylio Virus (Rgv) Iridovirus Dutpase (Dut) Early Viral Gene Protein Synthesis Inhibitor Dna Replication Inhibitor
DepartmentChinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
Subject AreaVirology
DOI10.1016/j.virusres.2006.08.007
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Indexed BySCI
Language英语
WOS Research AreaVirology
WOS SubjectVirology
WOS IDWOS:000244017300003
WOS KeywordINFECTIOUS-ANEMIA VIRUS ; FISH CELL-LINE ; FROG VIRUS-3 ; CRYSTAL-STRUCTURE ; DEOXYURIDINE TRIPHOSPHATASE ; MACROMOLECULAR-SYNTHESIS ; REPLICATION ; IRIDOVIRUS ; PROTEIN ; TYPE-1
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Cited Times:33[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/8706
Collection期刊论文
Corresponding AuthorZhang, QY, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
AffiliationChinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
Recommended Citation
GB/T 7714
Zhao, Zhe,Ke, Fei,Gui, Jianfang,et al. Characterization of an early gene encoding for dUTPase in Rana grylio virus[J]. VIRUS RESEARCH,2007,123(2):128-137.
APA Zhao, Zhe,Ke, Fei,Gui, Jianfang,Zhang, Qiya,&Zhang, QY, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China.(2007).Characterization of an early gene encoding for dUTPase in Rana grylio virus.VIRUS RESEARCH,123(2),128-137.
MLA Zhao, Zhe,et al."Characterization of an early gene encoding for dUTPase in Rana grylio virus".VIRUS RESEARCH 123.2(2007):128-137.
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