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Characterization of porcine P58IPK gene and its up-regulation after H1N1 or H3N2 influenza virus infection
Pengfei Jianga, b, c, 1,
Junge Wena, b, c, 1,
Hao Songa, b, c,
Xinyu Chena, b, c,
Yan Suna, b, c,
Xuexi Huod, Corresponding author contact information, E-mail the corresponding author,
Deli Zhanga, b, c, Corresponding author contact information, E-mail the corresponding author
a MOA Key Laboratory of Animal Biotechnology of National Ministry of Agriculture, Institute of Veterinary Immunology, Northwest A&F University, Yangling, 712100, Xi?an City, Shaanxi Province, PR China
b Research Laboratory of Virology, Immunology & Bioinformatics, Division of Veterinary Microbiology & Virology, Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, 712100, Xi?an City, Shaanxi Province, PR China
c Investigation Group of Molecular Virology, Immunology, Oncology & Systems Biology, Center for Bioinformatics, Northwest A&F University, Yangling, 712100, Xi?an City, Shaanxi Province, PR China
d Department of Agricultural Economics, College of Economics and Management, Northwest A&F University, Yangling 712100, Xi?an City, Shaanxi, PR China
Abstract
Background
The 58-kDa inhibitor of the interferon-induced double-stranded RNA-activated protein kinase (P58IPK) is a cellular protein that is activated during influenza virus infection. Although the function of human P58IPK has been studied for a long time, porcine P58IPK (pP58IPK) has little been studied except for its cloning.
Objective
In this study, we aimed to investigate the characteristics of the pP58IPK gene, determine its subcellular localization, and find its expression change during H1N1 or H3N2 infection.
Study design
First, the sequence and structure of pP58IPK were analyzed. Second, pP58IPK gene was cloned into pEGFP-N1 and pEGFP-C1 vectors, respectively, which were transfected into cells to determine its subcellular localization. Third, Lung tissues of piglets from H1N1 infected, H3N2 infected and control groups were analyzed using histopathology, real-time PCR, and immunohistochemistry.
Results
The sequence and structure of pP58IPK was highly similar to the counterpart of human. pP58IPK protein distributed only in the cytoplasm. Lung tissues of piglets infected by H1N1 or H3N2 appeared obvious pathological changes, and the expression of pP58IPK in both mRNA and protein level was up-regulated by approximate 1.5-fold in piglets infected by H1N1 or H3N2 comparing with control piglets.
Conclusions
We analyzed the characteristics of the pP58IPK gene, constructed a phylogenetic tree, determined its subcellular localization, and investigated its expression changes during H1N1 or H3N2 infection. The fundamental data accumulated in this study provides a potential medical model for investigating the function of P58IPK during influenza A viruses infection.
Pengfei Jianga, b, c, 1,
Junge Wena, b, c, 1,
Hao Songa, b, c,
Xinyu Chena, b, c,
Yan Suna, b, c,
Xuexi Huod, Corresponding author contact information, E-mail the corresponding author,
Deli Zhanga, b, c, Corresponding author contact information, E-mail the corresponding author
a MOA Key Laboratory of Animal Biotechnology of National Ministry of Agriculture, Institute of Veterinary Immunology, Northwest A&F University, Yangling, 712100, Xi?an City, Shaanxi Province, PR China
b Research Laboratory of Virology, Immunology & Bioinformatics, Division of Veterinary Microbiology & Virology, Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, 712100, Xi?an City, Shaanxi Province, PR China
c Investigation Group of Molecular Virology, Immunology, Oncology & Systems Biology, Center for Bioinformatics, Northwest A&F University, Yangling, 712100, Xi?an City, Shaanxi Province, PR China
d Department of Agricultural Economics, College of Economics and Management, Northwest A&F University, Yangling 712100, Xi?an City, Shaanxi, PR China
Abstract
Background
The 58-kDa inhibitor of the interferon-induced double-stranded RNA-activated protein kinase (P58IPK) is a cellular protein that is activated during influenza virus infection. Although the function of human P58IPK has been studied for a long time, porcine P58IPK (pP58IPK) has little been studied except for its cloning.
Objective
In this study, we aimed to investigate the characteristics of the pP58IPK gene, determine its subcellular localization, and find its expression change during H1N1 or H3N2 infection.
Study design
First, the sequence and structure of pP58IPK were analyzed. Second, pP58IPK gene was cloned into pEGFP-N1 and pEGFP-C1 vectors, respectively, which were transfected into cells to determine its subcellular localization. Third, Lung tissues of piglets from H1N1 infected, H3N2 infected and control groups were analyzed using histopathology, real-time PCR, and immunohistochemistry.
Results
The sequence and structure of pP58IPK was highly similar to the counterpart of human. pP58IPK protein distributed only in the cytoplasm. Lung tissues of piglets infected by H1N1 or H3N2 appeared obvious pathological changes, and the expression of pP58IPK in both mRNA and protein level was up-regulated by approximate 1.5-fold in piglets infected by H1N1 or H3N2 comparing with control piglets.
Conclusions
We analyzed the characteristics of the pP58IPK gene, constructed a phylogenetic tree, determined its subcellular localization, and investigated its expression changes during H1N1 or H3N2 infection. The fundamental data accumulated in this study provides a potential medical model for investigating the function of P58IPK during influenza A viruses infection.