Expression regulation and functional characterization of a novel interferon inducible gene Gig2 and its promoter
Grass carp hemorrhagic virus (GCHV)-induced gene 2 (Gig2) is a novel gene previously identified from UV-inactivated GCHV-treated Carassius auratus blastulae embryonic (CAB) cells, suggesting that it should play a pivotal role in the interferon (IFN) antiviral response. In this study, a polyclonal an...
Published in: | Molecular Immunology |
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Main Authors: | , , , , , , |
Format: | Article in Journal/Newspaper |
Language: | English |
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2009
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Online Access: | http://ir.ihb.ac.cn/handle/152342/7564 https://doi.org/10.1016/j.molimm.2009.05.183 |
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openpolar |
institution |
Open Polar |
collection |
Institute of Hydrobiology, Chinese Academy of Sciences: IHB OpenIR |
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ftchinacadsciihb |
language |
English |
topic |
Gig2 Interferon Interferon-stimulated Gene (Isg) Irf7 Poly i:c Promoter Interferon-stimulated Response Elements Expression Regulation Biochemistry & Molecular Biology Immunology Science & Technology Life Sciences & Biomedicine TROUT ONCORHYNCHUS-MYKISS NF-KAPPA-B CARP-HEMORRHAGE-VIRUS RAINBOW-TROUT ATLANTIC SALMON PARALICHTHYS-OLIVACEUS CAB CELLS MX GENE MOLECULAR CHARACTERIZATION JAPANESE FLOUNDER |
spellingShingle |
Gig2 Interferon Interferon-stimulated Gene (Isg) Irf7 Poly i:c Promoter Interferon-stimulated Response Elements Expression Regulation Biochemistry & Molecular Biology Immunology Science & Technology Life Sciences & Biomedicine TROUT ONCORHYNCHUS-MYKISS NF-KAPPA-B CARP-HEMORRHAGE-VIRUS RAINBOW-TROUT ATLANTIC SALMON PARALICHTHYS-OLIVACEUS CAB CELLS MX GENE MOLECULAR CHARACTERIZATION JAPANESE FLOUNDER Jiang, Jun Zhang, Yi-Bing Li, Shun Yu, Fei-Fei Sun, Fan Gui, Jian-Fang Zhang, YB, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, 7 Donghu S Rd, Wuhan 430072, Peoples R China Expression regulation and functional characterization of a novel interferon inducible gene Gig2 and its promoter |
topic_facet |
Gig2 Interferon Interferon-stimulated Gene (Isg) Irf7 Poly i:c Promoter Interferon-stimulated Response Elements Expression Regulation Biochemistry & Molecular Biology Immunology Science & Technology Life Sciences & Biomedicine TROUT ONCORHYNCHUS-MYKISS NF-KAPPA-B CARP-HEMORRHAGE-VIRUS RAINBOW-TROUT ATLANTIC SALMON PARALICHTHYS-OLIVACEUS CAB CELLS MX GENE MOLECULAR CHARACTERIZATION JAPANESE FLOUNDER |
description |
Grass carp hemorrhagic virus (GCHV)-induced gene 2 (Gig2) is a novel gene previously identified from UV-inactivated GCHV-treated Carassius auratus blastulae embryonic (CAB) cells, suggesting that it should play a pivotal role in the interferon (IFN) antiviral response. In this study, a polyclonal anti-Gig2 antiserum was generated and used to study the inductive expression pattern by Western blot analysis, showing no basal expression in normal CAB cells but a significant up-regulation upon UV-inactivated GCHV, polyinosinic:polycytidylic acid (Poly I:Q and recombinant IFN (rIFN). However, constitutive expression of Gig2 is observed in all tested tissues from grass carp (Ctenopharyngodon idellus), and Poly I:C injection increases the relative amount of Gig2 protein in skin, spleen, trunk kidney, gill, hindgut and thymus. Moreover, the genomic sequence covering the whole Gig2 ORF and the upstream promoter region were amplified by genomic walking. Significantly, the Gig2 promoter contains three IFN-stimulated response elements (ISREs), nine GAAA/TfTC motifs and five gamma-IFN activating sites (GAS), which are the characteristics of genes responsive to both type I IFN and type 11 IFN. Subsequently, the complete Gig2 promoter sequence was cloned into pGL3-Basic vector, and its activity was measured by luciferase assays in the transfected CAB cells. The Gig2 promoter-driven construct is highly induced in CAB cells after treatment with Poly I:C or rIFN, and the functional capability is dependent on IFN regulatory factor 7 (IRF7), because its activity can be stimulated by IRF7. Collectively, the data provide strong evidence that Gig2 is indeed a novel IFN inducible gene and its expression is likely dependent on IRF7 upon Poly I:C or IFN. (C) 2009 Elsevier Ltd. All rights reserved. Grass carp hemorrhagic virus (GCHV)-induced gene 2 (Gig2) is a novel gene previously identified from UV-inactivated GCHV-treated Carassius auratus blastulae embryonic (CAB) cells, suggesting that it should play a pivotal role in the interferon (IFN) antiviral response. In this study, a polyclonal anti-Gig2 antiserum was generated and used to study the inductive expression pattern by Western blot analysis, showing no basal expression in normal CAB cells but a significant up-regulation upon UV-inactivated GCHV, polyinosinic:polycytidylic acid (Poly I:Q and recombinant IFN (rIFN). However, constitutive expression of Gig2 is observed in all tested tissues from grass carp (Ctenopharyngodon idellus), and Poly I:C injection increases the relative amount of Gig2 protein in skin, spleen, trunk kidney, gill, hindgut and thymus. Moreover, the genomic sequence covering the whole Gig2 ORF and the upstream promoter region were amplified by genomic walking. Significantly, the Gig2 promoter contains three IFN-stimulated response elements (ISREs), nine GAAA/TfTC motifs and five gamma-IFN activating sites (GAS), which are the characteristics of genes responsive to both type I IFN and type 11 IFN. Subsequently, the complete Gig2 promoter sequence was cloned into pGL3-Basic vector, and its activity was measured by luciferase assays in the transfected CAB cells. The Gig2 promoter-driven construct is highly induced in CAB cells after treatment with Poly I:C or rIFN, and the functional capability is dependent on IFN regulatory factor 7 (IRF7), because its activity can be stimulated by IRF7. Collectively, the data provide strong evidence that Gig2 is indeed a novel IFN inducible gene and its expression is likely dependent on IRF7 upon Poly I:C or IFN. (C) 2009 Elsevier Ltd. All rights reserved. |
format |
Article in Journal/Newspaper |
author |
Jiang, Jun Zhang, Yi-Bing Li, Shun Yu, Fei-Fei Sun, Fan Gui, Jian-Fang Zhang, YB, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, 7 Donghu S Rd, Wuhan 430072, Peoples R China |
author_facet |
Jiang, Jun Zhang, Yi-Bing Li, Shun Yu, Fei-Fei Sun, Fan Gui, Jian-Fang Zhang, YB, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, 7 Donghu S Rd, Wuhan 430072, Peoples R China |
author_sort |
Jiang, Jun |
title |
Expression regulation and functional characterization of a novel interferon inducible gene Gig2 and its promoter |
title_short |
Expression regulation and functional characterization of a novel interferon inducible gene Gig2 and its promoter |
title_full |
Expression regulation and functional characterization of a novel interferon inducible gene Gig2 and its promoter |
title_fullStr |
Expression regulation and functional characterization of a novel interferon inducible gene Gig2 and its promoter |
title_full_unstemmed |
Expression regulation and functional characterization of a novel interferon inducible gene Gig2 and its promoter |
title_sort |
expression regulation and functional characterization of a novel interferon inducible gene gig2 and its promoter |
publishDate |
2009 |
url |
http://ir.ihb.ac.cn/handle/152342/7564 https://doi.org/10.1016/j.molimm.2009.05.183 |
genre |
Atlantic salmon |
genre_facet |
Atlantic salmon |
op_relation |
MOLECULAR IMMUNOLOGY Jiang, Jun; Zhang, Yi-Bing; Li, Shun; Yu, Fei-Fei; Sun, Fan; Gui, Jian-Fang.Expression regulation and functional characterization of a novel interferon inducible gene Gig2 and its promoter,MOLECULAR IMMUNOLOGY,2009,46(15):3131-3140 http://ir.ihb.ac.cn/handle/152342/7564 doi:10.1016/j.molimm.2009.05.183 |
op_doi |
https://doi.org/10.1016/j.molimm.2009.05.183 |
container_title |
Molecular Immunology |
container_volume |
46 |
container_issue |
15 |
container_start_page |
3131 |
op_container_end_page |
3140 |
_version_ |
1766363637387100160 |
spelling |
ftchinacadsciihb:oai:ir.ihb.ac.cn:152342/7564 2023-05-15T15:33:10+02:00 Expression regulation and functional characterization of a novel interferon inducible gene Gig2 and its promoter Jiang, Jun Zhang, Yi-Bing Li, Shun Yu, Fei-Fei Sun, Fan Gui, Jian-Fang Zhang, YB, Chinese Acad Sci, Grad Sch, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, 7 Donghu S Rd, Wuhan 430072, Peoples R China 2009-09-01 http://ir.ihb.ac.cn/handle/152342/7564 https://doi.org/10.1016/j.molimm.2009.05.183 英语 eng MOLECULAR IMMUNOLOGY Jiang, Jun; Zhang, Yi-Bing; Li, Shun; Yu, Fei-Fei; Sun, Fan; Gui, Jian-Fang.Expression regulation and functional characterization of a novel interferon inducible gene Gig2 and its promoter,MOLECULAR IMMUNOLOGY,2009,46(15):3131-3140 http://ir.ihb.ac.cn/handle/152342/7564 doi:10.1016/j.molimm.2009.05.183 Gig2 Interferon Interferon-stimulated Gene (Isg) Irf7 Poly i:c Promoter Interferon-stimulated Response Elements Expression Regulation Biochemistry & Molecular Biology Immunology Science & Technology Life Sciences & Biomedicine TROUT ONCORHYNCHUS-MYKISS NF-KAPPA-B CARP-HEMORRHAGE-VIRUS RAINBOW-TROUT ATLANTIC SALMON PARALICHTHYS-OLIVACEUS CAB CELLS MX GENE MOLECULAR CHARACTERIZATION JAPANESE FLOUNDER Article 期刊论文 2009 ftchinacadsciihb https://doi.org/10.1016/j.molimm.2009.05.183 2019-07-01T11:26:30Z Grass carp hemorrhagic virus (GCHV)-induced gene 2 (Gig2) is a novel gene previously identified from UV-inactivated GCHV-treated Carassius auratus blastulae embryonic (CAB) cells, suggesting that it should play a pivotal role in the interferon (IFN) antiviral response. In this study, a polyclonal anti-Gig2 antiserum was generated and used to study the inductive expression pattern by Western blot analysis, showing no basal expression in normal CAB cells but a significant up-regulation upon UV-inactivated GCHV, polyinosinic:polycytidylic acid (Poly I:Q and recombinant IFN (rIFN). However, constitutive expression of Gig2 is observed in all tested tissues from grass carp (Ctenopharyngodon idellus), and Poly I:C injection increases the relative amount of Gig2 protein in skin, spleen, trunk kidney, gill, hindgut and thymus. Moreover, the genomic sequence covering the whole Gig2 ORF and the upstream promoter region were amplified by genomic walking. Significantly, the Gig2 promoter contains three IFN-stimulated response elements (ISREs), nine GAAA/TfTC motifs and five gamma-IFN activating sites (GAS), which are the characteristics of genes responsive to both type I IFN and type 11 IFN. Subsequently, the complete Gig2 promoter sequence was cloned into pGL3-Basic vector, and its activity was measured by luciferase assays in the transfected CAB cells. The Gig2 promoter-driven construct is highly induced in CAB cells after treatment with Poly I:C or rIFN, and the functional capability is dependent on IFN regulatory factor 7 (IRF7), because its activity can be stimulated by IRF7. Collectively, the data provide strong evidence that Gig2 is indeed a novel IFN inducible gene and its expression is likely dependent on IRF7 upon Poly I:C or IFN. (C) 2009 Elsevier Ltd. All rights reserved. Grass carp hemorrhagic virus (GCHV)-induced gene 2 (Gig2) is a novel gene previously identified from UV-inactivated GCHV-treated Carassius auratus blastulae embryonic (CAB) cells, suggesting that it should play a pivotal role in the interferon (IFN) antiviral response. In this study, a polyclonal anti-Gig2 antiserum was generated and used to study the inductive expression pattern by Western blot analysis, showing no basal expression in normal CAB cells but a significant up-regulation upon UV-inactivated GCHV, polyinosinic:polycytidylic acid (Poly I:Q and recombinant IFN (rIFN). However, constitutive expression of Gig2 is observed in all tested tissues from grass carp (Ctenopharyngodon idellus), and Poly I:C injection increases the relative amount of Gig2 protein in skin, spleen, trunk kidney, gill, hindgut and thymus. Moreover, the genomic sequence covering the whole Gig2 ORF and the upstream promoter region were amplified by genomic walking. Significantly, the Gig2 promoter contains three IFN-stimulated response elements (ISREs), nine GAAA/TfTC motifs and five gamma-IFN activating sites (GAS), which are the characteristics of genes responsive to both type I IFN and type 11 IFN. Subsequently, the complete Gig2 promoter sequence was cloned into pGL3-Basic vector, and its activity was measured by luciferase assays in the transfected CAB cells. The Gig2 promoter-driven construct is highly induced in CAB cells after treatment with Poly I:C or rIFN, and the functional capability is dependent on IFN regulatory factor 7 (IRF7), because its activity can be stimulated by IRF7. Collectively, the data provide strong evidence that Gig2 is indeed a novel IFN inducible gene and its expression is likely dependent on IRF7 upon Poly I:C or IFN. (C) 2009 Elsevier Ltd. All rights reserved. Article in Journal/Newspaper Atlantic salmon Institute of Hydrobiology, Chinese Academy of Sciences: IHB OpenIR Molecular Immunology 46 15 3131 3140 |