title: Characterization of porcine P58IPK gene and its up-regulation after H1N1 or H3N2 influenza virus infection.
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.
  
  
  METHODS
  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.
authors.affiliation: 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.
authors.forename: Pengfei
authors.initials: P
authors.lastname: Jiang
authors.forename: Junge
authors.initials: J
authors.lastname: Wen
authors.forename: Hao
authors.initials: H
authors.lastname: Song
authors.forename: Xinyu
authors.initials: X
authors.lastname: Chen
authors.forename: Yan
authors.initials: Y
authors.lastname: Sun
authors.forename: Xuexi
authors.initials: X
authors.lastname: Huo
authors.forename: Deli
authors.initials: D
authors.lastname: Zhang
mesh_terms: D000818:Animals; D016000:Cluster Analysis; D050956:HSP40 Heat-Shock Proteins / Q000096:biosynthesis* / Q000235:genetics; D006651:Histocytochemistry; D054884:Host-Pathogen Interactions; D007150:Immunohistochemistry; D053118:Influenza A Virus, H1N1 Subtype / Q000254:growth & development*; D053122:Influenza A Virus, H3N2 Subtype / Q000254:growth & development*; D008168:Lung / Q000821:virology; D008969:Molecular Sequence Data; D009976:Orthomyxoviridae Infections / Q000276:immunology / Q000662:veterinary* / Q000821:virology; D010802:Phylogeny; D060888:Real-Time Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D013552:Swine; D013553:Swine Diseases / Q000276:immunology / Q000821:virology*; D015854:Up-Regulation
chemical_list: D050956:HSP40 Heat-Shock Proteins
keywords: C(T); ER; H1N1; H3N2; HE; M(w); ORF; Open Reading Frame; P58(IPK); PKR; Porcine P58(IPK); SUVEC; Subcellular localization; TPR; Up-regulation; dsRNA dependent protein kinase; eIF2a; endoplasmic reticulum; hematoxylin–eosin; isoelectric point; molecular weight; p(I); pP58(IPK); porcine P58IPK; swine umbilical vein endothelial cell line; tetratricopeptide repeat; the 58-kDa inhibitor of the interferon-induced double-stranded RNA-activated protein kinase; the eukaryotic translation initiation factor 2 alpha subunit; threshold cycle