title: Genome-wide RNAi screen identifies human host factors crucial for influenza virus replication.
abstract: Influenza A virus, being responsible for seasonal epidemics and reoccurring pandemics, represents a worldwide threat to public health. High mutation rates facilitate the generation of viral escape mutants, rendering vaccines and drugs directed against virus-encoded targets potentially ineffective. In contrast, targeting host cell determinants temporarily dispensable for the host but crucial for virus replication could prevent viral escape. Here we report the discovery of 287 human host cell genes influencing influenza A virus replication in a genome-wide RNA interference (RNAi) screen. Using an independent assay we confirmed 168 hits (59%) inhibiting either the endemic H1N1 (119 hits) or the current pandemic swine-origin (121 hits) influenza A virus strains, with an overlap of 60%. Notably, a subset of these common hits was also essential for replication of a highly pathogenic avian H5N1 strain. In-depth analyses of several factors provided insights into their infection stage relevance. Notably, SON DNA binding protein (SON) was found to be important for normal trafficking of influenza virions to late endosomes early in infection. We also show that a small molecule inhibitor of CDC-like kinase 1 (CLK1) reduces influenza virus replication by more than two orders of magnitude, an effect connected with impaired splicing of the viral M2 messenger RNA. Furthermore, influenza-virus-infected p27(-/-) (cyclin-dependent kinase inhibitor 1B; Cdkn1b) mice accumulated significantly lower viral titres in the lung, providing in vivo evidence for the importance of this gene. Thus, our results highlight the potency of genome-wide RNAi screening for the dissection of virus-host interactions and the identification of drug targets for a broad range of influenza viruses.
authors.affiliation: Molecular Biology Department, Max Planck Institute for Infection Biology, Charitéplatz 1, 10117 Berlin, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Karlas
authors.forename: Nikolaus
authors.initials: N
authors.lastname: Machuy
authors.forename: Yujin
authors.initials: Y
authors.lastname: Shin
authors.forename: Klaus-Peter
authors.initials: KP
authors.lastname: Pleissner
authors.forename: Anita
authors.initials: A
authors.lastname: Artarini
authors.forename: Dagmar
authors.initials: D
authors.lastname: Heuer
authors.forename: Daniel
authors.initials: D
authors.lastname: Becker
authors.forename: Hany
authors.initials: H
authors.lastname: Khalil
authors.forename: Lesley A
authors.initials: LA
authors.lastname: Ogilvie
authors.forename: Simone
authors.initials: S
authors.lastname: Hess
authors.forename: André P
authors.initials: AP
authors.lastname: Mäurer
authors.forename: Elke
authors.initials: E
authors.lastname: Müller
authors.forename: Thorsten
authors.initials: T
authors.lastname: Wolff
authors.forename: Thomas
authors.initials: T
authors.lastname: Rudel
authors.forename: Thomas F
authors.initials: TF
authors.lastname: Meyer
mesh_terms: D000818:Animals; D001685:Biological Factors / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D002478:Cells, Cultured; D002642:Chick Embryo; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D004847:Epithelial Cells / Q000821:virology; D015894:Genome, Human / Q000235:genetics; D054884:Host-Pathogen Interactions / Q000235:genetics / Q000502:physiology; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000145:classification / Q000254:growth & development*; D007251:Influenza, Human / Q000235:genetics* / Q000821:virology*; D008168:Lung / Q000166:cytology; D051379:Mice; D008810:Mice, Inbred C57BL; D017346:Protein Serine-Threonine Kinases / Q000235:genetics; D011505:Protein-Tyrosine Kinases / Q000235:genetics; D034622:RNA Interference; D014779:Virus Replication / Q000502:physiology*
chemical_list: D001685:Biological Factors; C493961:Cdkn1b protein, mouse; D050760:Cyclin-Dependent Kinase Inhibitor p27; C066780:Clk dual-specificity kinases; D011505:Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases