Identification of two ISG15 homologues involved in host immune response against RGNNV in Asian seabass (Lates calcarifer)

Interferon Stimulated Gene (ISG)15 is a ubiquitin-like protein that is induced upon viral infections. Our study reports the identification of two homologues of ISG15 in the Asian seabass designated LcISG15A and LcISG15B. The cloned LcISG15A cDNA fragment contained a 474 bp ORF encoding a 157 amino a...

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Main Authors: R.S. Krishna Priya, Avinash Premraj, K.C. Sivakumar, T.P. Sajeevan
Format: Article
Language:English
Published: Elsevier 2022-12-01
Series:Fish and Shellfish Immunology Reports
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2667011922000056
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author R.S. Krishna Priya
Avinash Premraj
K.C. Sivakumar
T.P. Sajeevan
author_facet R.S. Krishna Priya
Avinash Premraj
K.C. Sivakumar
T.P. Sajeevan
author_sort R.S. Krishna Priya
collection DOAJ
description Interferon Stimulated Gene (ISG)15 is a ubiquitin-like protein that is induced upon viral infections. Our study reports the identification of two homologues of ISG15 in the Asian seabass designated LcISG15A and LcISG15B. The cloned LcISG15A cDNA fragment contained a 474 bp ORF encoding a 157 amino acid protein whereas LcISG15B featured a 498 bp ORF encoding a slightly longer protein of 165 amino acids. Both proteins featured the two tandem ubiquitin-like domains and the C-terminal LRGG motif characteristic of ISG15. The LcISG15B protein has a 10-amino acid C-terminal extension after the LRGG motif. Molecular docking studies revealed that LcISG15A showed more conformational variability of the ubiquitin domains and catalytic function than LcISG15B. The Lates ISG15A and ISG15B genes, reside close in the genome, share the same basic structure with two exons and an intron, but only the second exon encoding the protein. These genes also featured the IFN-stimulatory response elements (ISRE) in the promoter region and ATTTA instability motif in the 3′ UTR region. Leukocyte-rich organs such as the head kidney, heart, spleen, and gill showed higher levels of ISG15A and ISG15B basal expression. Poly (I:C) injection rapidly upregulated the transcription of both the ISG15 genes in these tissues in Lates. In-vivo viral infection by red-spotted grouper nervous necrosis virus also induced upregulation of ISG15 genes in the head kidney, spleen, heart and gill. These findings indicate that the two ISG15 homologues may play a crucial role in innate antiviral immunity and could be used to improve prophylactic strategies and develop species-specific immunological tools for Lates calcarifer.
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spelling doaj.art-1bd975c55fec471aa658ed2402d0680a2022-12-22T04:23:19ZengElsevierFish and Shellfish Immunology Reports2667-01192022-12-013100054Identification of two ISG15 homologues involved in host immune response against RGNNV in Asian seabass (Lates calcarifer)R.S. Krishna Priya0Avinash Premraj1K.C. Sivakumar2T.P. Sajeevan3National Centre for Aquatic Animal Health, Cochin University of Science and Technology, Fine Arts Avenue, Kochi, 682 016, Kerala, IndiaCamel Biotechnology Centre, Presidential Camels and Camel Racing Affairs Centre, Department of the President's Affairs, PO Box 17292, Al Ain, United Arab EmiratesNational Centre for Aquatic Animal Health, Cochin University of Science and Technology, Fine Arts Avenue, Kochi, 682 016, Kerala, India; Rajiv Gandhi Centre for Biotechnology, Poojappura, Thiruvananthapuram, Kerala 695 014 IndiaNational Centre for Aquatic Animal Health, Cochin University of Science and Technology, Fine Arts Avenue, Kochi, 682 016, Kerala, India; Corresponding author.Interferon Stimulated Gene (ISG)15 is a ubiquitin-like protein that is induced upon viral infections. Our study reports the identification of two homologues of ISG15 in the Asian seabass designated LcISG15A and LcISG15B. The cloned LcISG15A cDNA fragment contained a 474 bp ORF encoding a 157 amino acid protein whereas LcISG15B featured a 498 bp ORF encoding a slightly longer protein of 165 amino acids. Both proteins featured the two tandem ubiquitin-like domains and the C-terminal LRGG motif characteristic of ISG15. The LcISG15B protein has a 10-amino acid C-terminal extension after the LRGG motif. Molecular docking studies revealed that LcISG15A showed more conformational variability of the ubiquitin domains and catalytic function than LcISG15B. The Lates ISG15A and ISG15B genes, reside close in the genome, share the same basic structure with two exons and an intron, but only the second exon encoding the protein. These genes also featured the IFN-stimulatory response elements (ISRE) in the promoter region and ATTTA instability motif in the 3′ UTR region. Leukocyte-rich organs such as the head kidney, heart, spleen, and gill showed higher levels of ISG15A and ISG15B basal expression. Poly (I:C) injection rapidly upregulated the transcription of both the ISG15 genes in these tissues in Lates. In-vivo viral infection by red-spotted grouper nervous necrosis virus also induced upregulation of ISG15 genes in the head kidney, spleen, heart and gill. These findings indicate that the two ISG15 homologues may play a crucial role in innate antiviral immunity and could be used to improve prophylactic strategies and develop species-specific immunological tools for Lates calcarifer.http://www.sciencedirect.com/science/article/pii/S2667011922000056Lates calcariferInterferon-stimulated geneRGNNVAntiviralTeleost
spellingShingle R.S. Krishna Priya
Avinash Premraj
K.C. Sivakumar
T.P. Sajeevan
Identification of two ISG15 homologues involved in host immune response against RGNNV in Asian seabass (Lates calcarifer)
Fish and Shellfish Immunology Reports
Lates calcarifer
Interferon-stimulated gene
RGNNV
Antiviral
Teleost
title Identification of two ISG15 homologues involved in host immune response against RGNNV in Asian seabass (Lates calcarifer)
title_full Identification of two ISG15 homologues involved in host immune response against RGNNV in Asian seabass (Lates calcarifer)
title_fullStr Identification of two ISG15 homologues involved in host immune response against RGNNV in Asian seabass (Lates calcarifer)
title_full_unstemmed Identification of two ISG15 homologues involved in host immune response against RGNNV in Asian seabass (Lates calcarifer)
title_short Identification of two ISG15 homologues involved in host immune response against RGNNV in Asian seabass (Lates calcarifer)
title_sort identification of two isg15 homologues involved in host immune response against rgnnv in asian seabass lates calcarifer
topic Lates calcarifer
Interferon-stimulated gene
RGNNV
Antiviral
Teleost
url http://www.sciencedirect.com/science/article/pii/S2667011922000056
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