title: Human miR-3145 inhibits influenza A viruses replication by targeting and silencing viral PB1 gene.
abstract: MicroRNAs (miRNAs) play an important role in the regulation of gene expression and are involved in many cellular processes including inhibition of viral replication in infected cells. In this study, three subtypes of influenza A viruses (pH1N1, H5N1 and H3N2) were analyzed to identify candidate human miRNAs targeting and silencing viral genes expression. Candidate human miRNAs were predicted by miRBase and RNAhybrid based on minimum free energy (MFE) and hybridization patterns between human miRNAs and viral target genes. In silico analysis presented 76 miRNAs targeting influenza A viruses, including 70 miRNAs that targeted specific subtypes (21 for pH1N1, 27 for H5N1 and 22 for H3N2) and 6 miRNAs (miR-216b, miR-3145, miR-3682, miR-4513, miR-4753 and miR-5693) that targeted multiple subtypes of influenza A viruses. Interestingly, miR-3145 is the only candidate miRNA targeting all three subtypes of influenza A viruses. The miR-3145 targets to PB1 encoding polymerase basic protein 1, which is the main component of the viral polymerase complex. The silencing effect of miR-3145 was validated by 3'-UTR reporter assay and inhibition of influenza viral replication in A549 cells. In 3'-UTR reporter assay, results revealed that miR-3145 triggered significant reduction of the luciferase activity. Moreover, expression of viral PB1 genes was also inhibited considerably (P value < 0.05) in viral infected cells expressing mimic miR-3145. In conclusion, this study demonstrated that human miR-3145 triggered silencing of viral PB1 genes and lead to inhibition of multiple subtypes of influenza viral replication. Therefore, hsa-miR-3145 might be useful for alternative treatment of influenza A viruses in the future.
authors.affiliation: Department of Biochemistry, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand.
authors.forename: Kritsada
authors.initials: K
authors.lastname: Khongnomnan
authors.affiliation: Department of Biochemistry, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand.
authors.forename: Jarika
authors.initials: J
authors.lastname: Makkoch
authors.affiliation: Research Affairs, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand.
authors.forename: Witthaya
authors.initials: W
authors.lastname: Poomipak
authors.affiliation: Center of excellent in clinical virology, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand.
authors.forename: Yong
authors.initials: Y
authors.lastname: Poovorawan
authors.affiliation: Department of Biochemistry, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand sp.medbiochemcu@gmail.com.
authors.forename: Sunchai
authors.initials: S
authors.lastname: Payungporn
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D020868:Gene Silencing / Q000187:drug effects; D005814:Genes, Viral / Q000187:drug effects; D053118:Influenza A Virus, H1N1 Subtype / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D053122:Influenza A Virus, H3N2 Subtype / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D053124:Influenza A Virus, H5N1 Subtype / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D029521:Alphainfluenzavirus / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D035683:MicroRNAs / Q000494:pharmacology*; D060888:Real-Time Polymerase Chain Reaction; D014764:Viral Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics; D014779:Virus Replication / Q000187:drug effects* / Q000235:genetics
chemical_list: D000998:Antiviral Agents; C000606724:MIRN3145 microRNA, human; D035683:MicroRNAs; D014764:Viral Proteins; C077346:influenza virus polymerase basic protein 1
keywords: influenza A virus; inhibition; miR-3145; miRNA; silencing; viral replication