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 health1. High mutation rates facilitate the generation

of viral escape mutants, rendering vaccines and drugs directed against virus-encoded targets potentially ineffective2. 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)3 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)4 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. Access through your institution Buy or subscribe This is a preview of

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large-scale RNAi screens. _Nature Methods_ 4, 847–849 (2007) Article  Google Scholar  Download references ACKNOWLEDGEMENTS The authors would like to thank M. Drabkina, G. Heins, D. Khalil

and D. Manntz for technical support, and K. Hoffmann and the experimental animal facility, headed by U. Klemm, for supporting mouse experiments. We also thank A. Pekosz for providing the

influenza luciferase reporter construct and S. Becker for the pandemic H1N1 influenza strain. This work was supported through the EU FP6 project RIGHT (LSHB-CT-2004-005276), ERA-Net

Pathogenomics (grant no. 0313938A) and a grant from FCI to T.F.M., and the RiNA network Berlin to T.R. and T.F.M. T.W. acknowledges German Government support via BMG (FSI program) and BMBF

(FluResearchNet). H.K. and A.A. are student fellows of the Egyptian Government and IMPRS-IDI, respectively. AUTHOR CONTRIBUTIONS A.K., N.M. and T.F.M. conceived the project; A.K., N.M., T.W.

and T.F.M. designed experiments; A.K., N.M., Y.S., A.A., D.H., D.B., H.K. and T.W. performed experiments; A.K., N.M., K.-P.P., S.H. and A.P.M. conducted bioinformatics; N.M., E.M., T.R. and

T.F.M. established the screening platform; A.K., N.M., D.H., D.B., L.A.O. and T.F.M. wrote the manuscript. AUTHOR INFORMATION Author notes * Simone Hess, Elke Müller & Thomas Rudel

Present address: Present addresses: Hannover Medical School, 30625 Hannover, Germany (S.H.); Exploratory Research Space, RWTH Aachen University, Templergraben 55, 52056 Aachen, Germany

(E.M.); Department of Microbiology, Biocenter, University of Würzburg, 97074 Würzburg, Germany (T.R.)., * Alexander Karlas and Nikolaus Machuy: These authors contributed equally to this

work. AUTHORS AND AFFILIATIONS * Molecular Biology Department,, Alexander Karlas, Nikolaus Machuy, Yujin Shin, Anita Artarini, Dagmar Heuer, Daniel Becker, Hany Khalil, Lesley A. Ogilvie, 

Simone Hess, André P. Mäurer, Elke Müller, Thomas Rudel & Thomas F. Meyer * Core Facility Bioinformatics, Max Planck Institute for Infection Biology, Charitéplatz 1, 10117 Berlin,

Germany , Klaus-Peter Pleissner * Robert Koch-Institute, Nordufer 20, 13353 Berlin, Germany , Thorsten Wolff Authors * Alexander Karlas View author publications You can also search for this

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Thomas F. Meyer. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no competing financial interests. SUPPLEMENTARY INFORMATION SUPPLEMENTARY INFORMATION This file contains

Supplementary Figures 1- 15 with Legends, a Supplementary Discussion, Supplementary References and Supplementary Tables 1-3. (PDF 1717 kb) POWERPOINT SLIDES POWERPOINT SLIDE FOR FIG. 1

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N., Shin, Y. _et al._ Genome-wide RNAi screen identifies human host factors crucial for influenza virus replication. _Nature_ 463, 818–822 (2010). https://doi.org/10.1038/nature08760

Download citation * Received: 10 October 2009 * Accepted: 17 December 2009 * Published: 17 January 2010 * Issue Date: 11 February 2010 * DOI: https://doi.org/10.1038/nature08760 SHARE THIS

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