title: Sus scrofa miR-204 and miR-4331 Negatively Regulate Swine H1N1/2009 Influenza A Virus Replication by Targeting Viral HA and NS, Respectively.
abstract: The prevalence of swine pandemic H1N1/2009 influenza A virus (SIV-H1N1/2009) in pigs has the potential to generate novel reassortant viruses, posing a great threat to human health. Cellular microRNAs (miRNAs) have been proven as promising small molecules for regulating influenza A virus replication by directly targeting viral genomic RNA. In this study, we predicted potentialSus scrofa(ssc-, swine) miRNAs targeting the genomic RNA of SIV-H1N1/2009 by RegRNA 2.0, and identified ssc-miR-204 and ssc-miR-4331 to target viral HA and NS respectively through dual-luciferase reporter assays. The messenger RNA (mRNA) levels of viral HA and NS were significantly suppressed when newborn pig trachea (NPTr) cells respectively overexpressed ssc-miR-204 and ssc-miR-4331 and were infected with SIV-H1N1/2009, whereas the suppression effect could be restored when respectively decreasing endogenous ssc-miR-204 and ssc-miR-4331 with inhibitors. Because of the importance of viral HA and NS in the life cycle of influenza A virus, ssc-miR-204 and ssc-miR-4331 exhibited an inhibition effect on SIV-H1N1/2009 replication. The antiviral effect was sequence-specific of SIV-H1N1/2009, for the target sites in HA and NS of H5N1 or H9N2 influenza A virus were not conserved. Furthermore, SIV-H1N1/2009 infection reversely downregulated the expression of ssc-miR-204 and ssc-miR-4331, which might facilitate the virus replication in the host. In summary, this work will provide us some important clues for controlling the prevalence of SIV-H1N1/2009 in pig populations.
authors.affiliation: State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China. zhangshishuo006@126.com.
authors.forename: Shishuo
authors.initials: S
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China. wangruifang29@163.com.
authors.forename: Ruifang
authors.initials: R
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China. shj741984661@126.com.
authors.forename: Huijuan
authors.initials: H
authors.lastname: Su
authors.affiliation: State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China. wangbiaoxiong@126.com.
authors.forename: Biaoxiong
authors.initials: B
authors.lastname: Wang
authors.affiliation: Department of Animal Science, Tibet Agricultural and Animal Husbandry College, Linzhi 860000, China. xzslsz@163.com.
authors.forename: Suolang
authors.initials: S
authors.lastname: Sizhu
authors.affiliation: State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China. lei460411658@163.com.
authors.forename: Zhixin
authors.initials: Z
authors.lastname: Lei
authors.affiliation: State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China. jml8328@126.com.|The Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, China. jml8328@126.com.
authors.forename: Meilin
authors.initials: M
authors.lastname: Jin
authors.affiliation: State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China. chenhch@mail.hzau.edu.cn.|The Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, China. chenhch@mail.hzau.edu.cn.
authors.forename: Huanchun
authors.initials: H
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China. caojiyue@mail.hzau.edu.cn.
authors.forename: Jiyue
authors.initials: J
authors.lastname: Cao
authors.affiliation: State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China. hbzhou@mail.hzau.edu.cn.|The Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, China. hbzhou@mail.hzau.edu.cn.
authors.forename: Hongbo
authors.initials: H
authors.lastname: Zhou
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D015153:Blotting, Western; D002478:Cells, Cultured; D015967:Gene Expression Regulation, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics* / Q000378:metabolism; D054884:Host-Pathogen Interactions / Q000235:genetics; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics* / Q000502:physiology; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D034421:Sus scrofa; D014132:Trachea / Q000166:cytology / Q000378:metabolism / Q000821:virology; D017361:Viral Nonstructural Proteins / Q000235:genetics* / Q000378:metabolism; D014779:Virus Replication / Q000235:genetics*
chemical_list: D019267:Hemagglutinin Glycoproteins, Influenza Virus; D035683:MicroRNAs; D012333:RNA, Messenger; D017361:Viral Nonstructural Proteins; D008156:Luciferases
keywords: RegRNA 2.0; Sus scrofa miR-204; Sus scrofa miR-4331; miRNA-virus interaction; viral genomic RNA; virus replication