A Role for H/ACA and C/D Small Nucleolar RNAs in Viral Replication
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We have employed gene-trap insertional mutagenesis to identify candidate genes whose disruption confer phenotypic resistance to lytic infection, in independent studies using 12 distinct viruses and several different cell lines. Analysis of >2,000 virus-resistant clones revealed >1,000 candidate host genes, approximately 20 % of which were disrupted in clones surviving separate infections with 2–6 viruses. Interestingly, there were 83 instances in which the insertional mutagenesis vector disrupted transcripts encoding H/ACA-class and C/D-class small nucleolar RNAs (SNORAs and SNORDs, respectively). Of these, 79 SNORAs and SNORDs reside within introns of 29 genes (predominantly protein-coding), while 4 appear to be independent transcription units. siRNA studies targeting candidate SNORA/Ds provided independent confirmation of their roles in infection when tested against cowpox virus, Dengue Fever virus, influenza A virus, human rhinovirus 16, herpes simplex virus 2, or respiratory syncytial virus. Significantly, eight of the nine SNORA/Ds targeted with siRNAs enhanced cellular resistance to multiple viruses suggesting widespread involvement of SNORA/Ds in virus–host interactions and/or virus-induced cell death.
KeywordsSNORA SNORD Virus Gene-trap siRNA Susceptibility
This work was supported by the Red and Bobby Buisson Foundation, and Public Health Service Small Business Innovation Research (SBIR) Grant AI084705 from the Division of AIDS, National Institute of Allergy and Infectious Diseases. DHR was supported by gifts from Maggie Chassman, the Red and Bobby Buisson Foundation, Inc., Zirus, Inc., and the Public Health Service. None of the funding sources influenced the study design, the collection, analysis of interpretation of data, the preparation of this manuscript, or the decision to submit the article for publication. We also thank Dr. H. Earl Ruley of Vanderbilt University for critical review of the manuscript, and Drs. Natalie McDonald and Thomas Hodge (Zirus, Inc.) for technical support.
Conflict of interest
The authors declare no conflict of interest.
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