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Molecular cloning and characterisation of a fish PKR-like gene from cultured CAB cells induced by UV-inactivated virus
Hu, CY; Zhang, YB; Huang, GP; Zhang, QY; Gui, HF; Gui, HF, Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Wuhan Ctr Dev Biol, Inst Hydrobiol, Wuhan 430072, Peoples R China
2004-10-01
Source PublicationFISH & SHELLFISH IMMUNOLOGY
ISSN1050-4648
Volume17Issue:4Pages:353-366
AbstractThe double-stranded-RNA-dependent protein kinase (PKR) is an important component in an antiviral defence pathway that is mediated by interferon (IFN) in vertebrates. Previously, some important IFN system genes had been identified from an IFN-producing CAB (crucian carp Carassius auratus blastulae embryonic) cells after treatment with UV-inactivated GCHV (grass carp haemorrhage virus). Here, a fish PKR-like gene, named CaPKR-like, is cloned and sequenced from the same virally infected CAB cells. It has 2192 base pairs in length with a largest open reading frame (ORF) encoding a protein of 513 amino acid residues. BLAST search reveals that the putative CaPKR-like protein is most homologous to human PKR and also has a high-level homology with all members of a family of eIF2alpha kinases. Structurally, CaPKR-like possesses a conserved C-terminal catalytic domain of eIF2alpha kinase family and the most similarity to mammalian PKRs. Within its N-terminus, there are no dsRNA-binding domains conserved in mammalian PKRs instead of two putative Z-DNA binding domains (Zalpha). Like mammalian PKRs, CaPKR-like had a very low level of constitutive expression in normal CAB cells but was up-regulated in response to active GCHV, UV-inactivated GCHV and CAB IFN, implying that the transcriptional activation of CaPKR-like by viral infection is mediated possibly by newly produced CAB IFN, which was further supported by using cycloheximide, a potent inhibitor of protein synthesis. The results together suggested that CaPKR-like was the first identified fish gene most similar to mammalian PKRs. (C) 2004 Elsevier Ltd. All rights reserved.; The double-stranded-RNA-dependent protein kinase (PKR) is an important component in an antiviral defence pathway that is mediated by interferon (IFN) in vertebrates. Previously, some important IFN system genes had been identified from an IFN-producing CAB (crucian carp Carassius auratus blastulae embryonic) cells after treatment with UV-inactivated GCHV (grass carp haemorrhage virus). Here, a fish PKR-like gene, named CaPKR-like, is cloned and sequenced from the same virally infected CAB cells. It has 2192 base pairs in length with a largest open reading frame (ORF) encoding a protein of 513 amino acid residues. BLAST search reveals that the putative CaPKR-like protein is most homologous to human PKR and also has a high-level homology with all members of a family of eIF2alpha kinases. Structurally, CaPKR-like possesses a conserved C-terminal catalytic domain of eIF2alpha kinase family and the most similarity to mammalian PKRs. Within its N-terminus, there are no dsRNA-binding domains conserved in mammalian PKRs instead of two putative Z-DNA binding domains (Zalpha). Like mammalian PKRs, CaPKR-like had a very low level of constitutive expression in normal CAB cells but was up-regulated in response to active GCHV, UV-inactivated GCHV and CAB IFN, implying that the transcriptional activation of CaPKR-like by viral infection is mediated possibly by newly produced CAB IFN, which was further supported by using cycloheximide, a potent inhibitor of protein synthesis. The results together suggested that CaPKR-like was the first identified fish gene most similar to mammalian PKRs. (C) 2004 Elsevier Ltd. All rights reserved.
SubtypeArticle
KeywordDouble-stranded-rna-dependent Protein Kinase Interferon Cab Cell Grass Carp Hemorrhage Virus Expression Antiviral-relevant Gene Immune-relevant Gene
DepartmentChinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Wuhan Ctr Dev Biol, Inst Hydrobiol, Wuhan 430072, Peoples R China; Nanchang Univ, Col Life Sci & Food Engn, Nanchang 330047, Peoples R China
Subject AreaFisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
DOI10.1016/j.fsi.2004.04.009
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Indexed BySCI
Language英语
WOS Research AreaFisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
WOS SubjectFisheries ; Immunology ; Marine & Freshwater Biology ; Veterinary Sciences
WOS IDWOS:000223586100006
WOS KeywordINDUCED PROTEIN-KINASE ; CARASSIUS-AURATUS L. ; Z-DNA ; EIF-2-ALPHA KINASE ; BINDING DOMAIN ; IN-VIVO ; INTERFERON ; RNA ; IDENTIFICATION ; INITIATION
Citation statistics
Cited Times:71[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/9440
Collection期刊论文
Corresponding AuthorGui, HF, Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Wuhan Ctr Dev Biol, Inst Hydrobiol, Wuhan 430072, Peoples R China
Affiliation1.Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Wuhan Ctr Dev Biol, Inst Hydrobiol, Wuhan 430072, Peoples R China
2.Nanchang Univ, Col Life Sci & Food Engn, Nanchang 330047, Peoples R China
Recommended Citation
GB/T 7714
Hu, CY,Zhang, YB,Huang, GP,et al. Molecular cloning and characterisation of a fish PKR-like gene from cultured CAB cells induced by UV-inactivated virus[J]. FISH & SHELLFISH IMMUNOLOGY,2004,17(4):353-366.
APA Hu, CY,Zhang, YB,Huang, GP,Zhang, QY,Gui, HF,&Gui, HF, Chinese Acad Sci, State Key Lab Freshwater Ecol & Biotechnol, Wuhan Ctr Dev Biol, Inst Hydrobiol, Wuhan 430072, Peoples R China.(2004).Molecular cloning and characterisation of a fish PKR-like gene from cultured CAB cells induced by UV-inactivated virus.FISH & SHELLFISH IMMUNOLOGY,17(4),353-366.
MLA Hu, CY,et al."Molecular cloning and characterisation of a fish PKR-like gene from cultured CAB cells induced by UV-inactivated virus".FISH & SHELLFISH IMMUNOLOGY 17.4(2004):353-366.
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