In Vivo RNAi Screening Identifies MDA5 as a Significant Contributor to the Cellular Defense against Influenza A Virus

被引:68
作者
Benitez, Asiel A. [1 ]
Panis, Maryline [1 ]
Xue, Jia [2 ]
Varble, Andrew [1 ]
Shim, Jaehee V. [1 ]
Frick, Amy L. [3 ]
Lopez, Carolina B. [2 ]
Sachs, David [4 ]
tenOever, Benjamin R. [1 ]
机构
[1] Icahn Sch Med Mt Sinai, Dept Microbiol, New York, NY 10029 USA
[2] Univ Penn, Sch Vet Med, Dept Pathobiol, Philadelphia, PA 19104 USA
[3] Icahn Sch Med Mt Sinai, Dept Neurosci, New York, NY 10029 USA
[4] Icahn Sch Med Mt Sinai, Dept Genet & Genom Sci, New York, NY 10029 USA
来源
CELL REPORTS | 2015年 / 11卷 / 11期
关键词
DOUBLE-STRANDED-RNA; WEST-NILE-VIRUS; RIG-I; NS1; PROTEIN; BETA-INTERFERON; MESSENGER-RNA; V-PROTEINS; RECOGNITION; ACTIVATION; INDUCTION;
D O I
10.1016/j.celrep.2015.05.032
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Responding to an influenza A virus (IAV) infection demands an effective intrinsic cellular defense strategy to slow replication. To identify contributing host factors to this defense, we exploited the host microRNA pathway to perform an in vivo RNAi screen. To this end, IAV, lacking a functional NS1 antagonist, was engineered to encode individual siRNAs against antiviral host genes in an effort to rescue attenuation. This screening platform resulted in the enrichment of strains targeting virus-activated transcription factors, specific antiviral effectors, and intracellular pattern recognition receptors (PRRs). Interestingly, in addition to RIG-I, the PRR for IAV, a virus with the capacity to silence MDA5 also emerged as a dominant strain in wild-type, but not in MDA5-deficient mice. Transcriptional profiling of infected knockout cells confirmed RIG-I to be the primary PRR for IAV but implicated MDA5 as a significant contributor to the cellular defense against influenza A virus.
引用
收藏
页码:1714 / 1726
页数:13
相关论文
共 52 条
[1]   The V proteins of paramyxoviruses bind the IFN-inducible RNA helicase, mda-5, and inhibit its activation of the IFN-β promoter [J].
Andrejeva, J ;
Childs, KS ;
Young, DF ;
Carlos, TS ;
Stock, N ;
Goodbourn, S ;
Randall, RE .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2004, 101 (49) :17264-17269
[2]   MicroRNAs: Target Recognition and Regulatory Functions [J].
Bartel, David P. .
CELL, 2009, 136 (02) :215-233
[3]   Derivation and characterization of Dicer- and microRNA-deficient human cells [J].
Bogerd, Hal P. ;
Whisnant, Adam W. ;
Kennedy, Edward M. ;
Flores, Omar ;
Cullen, Bryan R. .
RNA, 2014, 20 (06) :923-937
[4]   The IFITM Proteins Mediate Cellular Resistance to Influenza A H1N1 Virus, West Nile Virus, and Dengue Virus [J].
Brass, Abraham L. ;
Huang, I-Chueh ;
Benita, Yair ;
John, Sinu P. ;
Krishnan, Manoj N. ;
Feeley, Eric M. ;
Ryan, Bethany J. ;
Weyer, Jessica L. ;
van der Weyden, Louise ;
Fikrig, Erol ;
Adams, David J. ;
Xavier, Ramnik J. ;
Farzan, Michael ;
Elledge, Stephen J. .
CELL, 2009, 139 (07) :1243-1254
[5]   mda-5, but not RIG-I, is a common target for paramyxovirus V proteins [J].
Childs, Kay ;
Stock, Nicola ;
Ross, Craig ;
Andrejeva, Jelena ;
Hilton, Louise ;
Skinner, Michael ;
Randall, Richard ;
Goodbourn, Stephen .
VIROLOGY, 2007, 359 (01) :190-200
[6]   Influenza A Virus Utilizes Suboptimal Splicing to Coordinate the Timing of Infection [J].
Chua, Mark A. ;
Schmid, Sonja ;
Perez, Jasmine T. ;
Langlois, Ryan A. ;
tenOever, Benjamin R. .
CELL REPORTS, 2013, 3 (01) :23-29
[7]   RNase L Targets Distinct Sites in Influenza A Virus RNAs [J].
Cooper, Daphne A. ;
Banerjee, Shuvojit ;
Chakrabarti, Arindam ;
Garcia-Sastre, Adolfo ;
Hesselberth, Jay R. ;
Silverman, Robert H. ;
Barton, David J. .
JOURNAL OF VIROLOGY, 2015, 89 (05) :2764-2776
[8]   A recombinant influenza A virus expressing an RNA-binding-defective NS1 protein induces high levels of beta interferon and is attenuated in mice [J].
Donelan, NR ;
Basler, CF ;
García-Sastre, A .
JOURNAL OF VIROLOGY, 2003, 77 (24) :13257-13266
[9]   Functional Identification of Optimized RNAi Triggers Using a Massively Parallel Sensor Assay [J].
Fellmann, Christof ;
Zuber, Johannes ;
McJunkin, Katherine ;
Chang, Kenneth ;
Malone, Colin D. ;
Dickins, Ross A. ;
Xu, Qikai ;
Hengartner, Michael O. ;
Elledge, Stephen J. ;
Hannon, Gregory J. ;
Lowe, Scott W. .
MOLECULAR CELL, 2011, 41 (06) :733-746
[10]   TRIM25 RING-finger E3 ubiquitin ligase is essential for RIG-I-mediated antiviral activity [J].
Gack, Michaela U. ;
Shin, Young C. ;
Joo, Chul-Hyun ;
Urano, Tomohiko ;
Liang, Chengyu ;
Sun, Lijun ;
Takeuchi, Osamu ;
Akira, Shizuo ;
Chen, Zhijian ;
Inoue, Satoshi ;
Jung, Jae U. .
NATURE, 2007, 446 (7138) :916-U2