| 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 Publication | VIRUS RESEARCH
 |
| ISSN | 0168-1702 |
| Volume | 123Issue:2Pages:128-137 |
| Abstract | 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.; 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. |
| Subtype | Article |
| Keyword | Rana Grylio Virus (Rgv) Iridovirus Dutpase (Dut) Early Viral Gene Protein Synthesis Inhibitor Dna Replication Inhibitor |
| Department | Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China |
| Subject Area | Virology |
| DOI | 10.1016/j.virusres.2006.08.007 |
| WOS Headings | Science & Technology ; Life Sciences & Biomedicine |
| Indexed By | SCI |
| Language | 英语 |
| WOS Research Area | Virology |
| WOS Subject | Virology |
| WOS ID | WOS:000244017300003 |
| WOS Keyword | INFECTIOUS-ANEMIA VIRUS ; FISH CELL-LINE ; FROG VIRUS-3 ; CRYSTAL-STRUCTURE ; DEOXYURIDINE TRIPHOSPHATASE ; MACROMOLECULAR-SYNTHESIS ; REPLICATION ; IRIDOVIRUS ; PROTEIN ; TYPE-1 |
| Citation statistics | |
| Document Type | 期刊论文 |
| Identifier | http://ir.ihb.ac.cn/handle/152342/8706 |
| Collection | 期刊论文 |
| Corresponding Author | Zhang, QY, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China |
| Affiliation | Chinese 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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