title: Human microRNA-24 modulates highly pathogenic avian-origin H5N1 influenza A virus infection in A549 cells by targeting secretory pathway furin.
abstract: A common critical cellular event that many human enveloped viruses share is the requirement for proteolytic cleavage of the viral glycoprotein by furin in the host secretory pathway. For example, the furin-dependent proteolytic activation of highly pathogenic (HP) influenza A (infA) H5 and H7 haemagglutinin precursor (HA0) subtypes is critical for yielding fusion-competent infectious virions. In this study, we hypothesized that viral hijacking of the furin pathway by HP infA viruses to permit cleavage of HA0 could represent a novel molecular mechanism controlling the dynamic production of fusion-competent infectious virus particles during the viral life cycle. We explored the biological role of a newly identified furin-directed human microRNA, miR-24, in this process as a potential post-transcriptional regulator of the furin-mediated activation of HA0 and production of fusion-competent virions in the host secretory pathway. We report that miR-24 and furin are differentially expressed in human A549 cells infected with HP avian-origin infA H5N1. Using miR-24 mimics, we demonstrated a robust decrease in both furin mRNA levels and intracellular furin activity in A549 cells. Importantly, pretreatment of A549 cells with miR-24 mimicked these results: a robust decrease of H5N1 infectious virions and a complete block of H5N1 virus spread that was not observed in A549 cells infected with low-pathogenicity swine-origin infA H1N1 virus. Our results suggest that viral-specific downregulation of furin-directed microRNAs such as miR-24 during the life cycle of HP infA viruses may represent a novel regulatory mechanism that governs furin-mediated proteolytic activation of HA0 glycoproteins and production of infectious virions.
authors.affiliation: Department of Microbiology and Immunology, University of British Columbia, Vancouver, BC V6T 1Z4, Canada.
authors.forename: Emma-Kate
authors.initials: EK
authors.lastname: Loveday
authors.affiliation: National Centre for Foreign Animal Disease, Canadian Food Inspection Agency, Winnipeg, MB R3E 3M4, Canada.|Department of Microbiology and Immunology, University of British Columbia, Vancouver, BC V6T 1Z4, Canada.
authors.forename: Sandra
authors.initials: S
authors.lastname: Diederich
authors.affiliation: National Centre for Foreign Animal Disease, Canadian Food Inspection Agency, Winnipeg, MB R3E 3M4, Canada.
authors.forename: John
authors.initials: J
authors.lastname: Pasick
authors.affiliation: Department of Microbiology and Immunology, University of British Columbia, Vancouver, BC V6T 1Z4, Canada.
authors.forename: François
authors.initials: F
authors.lastname: Jean
mesh_terms: D000818:Animals; D001717:Birds / Q000821:virology; D002460:Cell Line; D003141:Communicable Diseases / Q000821:virology; D004285:Dogs; D045683:Furin / Q000235:genetics*; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics; D054884:Host-Pathogen Interactions / Q000235:genetics; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics*; D005585:Influenza in Birds / Q000821:virology; D007251:Influenza, Human / Q000821:virology*; D061985:Madin Darby Canine Kidney Cells; D035683:MicroRNAs / Q000235:genetics*; D012323:RNA Processing, Post-Transcriptional / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D055571:Secretory Pathway / Q000235:genetics*
chemical_list: D019267:Hemagglutinin Glycoproteins, Influenza Virus; C543845:MIRN24 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger; C109963:hemagglutinin, avian influenza A virus; D045683:Furin