title: RNA-based immunity terminates viral infection in adult Drosophila in the absence of viral suppression of RNA interference: characterization of viral small interfering RNA populations in wild-type and mutant flies.
abstract: Replication of viral RNA genomes in fruit flies and mosquitoes induces the production of virus-derived small interfering RNAs (siRNAs) to specifically reduce virus accumulation by RNA interference (RNAi). However, it is unknown whether the RNA-based antiviral immunity (RVI) is sufficiently potent to terminate infection in adult insects as occurs in cell culture. We show here that, in contrast to robust infection by Flock house virus (FHV), infection with an FHV mutant (FHVΔB2) unable to express its RNAi suppressor protein B2 was rapidly terminated in adult flies. FHVΔB2 replicated to high levels and induced high mortality rates in dicer-2 and argonaute-2 mutant flies that are RNAi defective, demonstrating that successful infection of adult Drosophila requires a virus-encoded activity to suppress RVI. Drosophila RVI may depend on the RNAi activity of viral siRNAs since efficient FHVΔB2 infection occurred in argonaute-2 and r2d2 mutant flies despite massive production of viral siRNAs. However, RVI appears to be insensitive to the relative abundance of viral siRNAs since FHVΔB2 infection was terminated in flies carrying a partial loss-of-function mutation in loquacious required for viral siRNA biogenesis. Deep sequencing revealed a low-abundance population of Dicer-2-dependent viral siRNAs accompanying FHVΔB2 infection arrest in RVI-competent flies that included an approximately equal ratio of positive and negative strands. Surprisingly, viral small RNAs became strongly biased for positive strands at later stages of infection in RVI-compromised flies due to genetic or viral suppression of RNAi. We propose that degradation of the asymmetrically produced viral positive-strand RNAs associated with abundant virus accumulation contributes to the positive-strand bias of viral small RNAs.
authors.affiliation: Department of Plant Pathology and Microbiology and Institute for Integrative Genome Biology and Program for Cell, Molecular, and Developmental Biology, University of California, Riverside, CA 92521, USA.
authors.forename: Yan-Hong
authors.initials: YH
authors.lastname: Han
authors.forename: Ying-Jun
authors.initials: YJ
authors.lastname: Luo
authors.forename: Qingfa
authors.initials: Q
authors.lastname: Wu
authors.forename: Juan
authors.initials: J
authors.lastname: Jovel
authors.forename: Xiao-Hong
authors.initials: XH
authors.lastname: Wang
authors.forename: Roghiyh
authors.initials: R
authors.lastname: Aliyari
authors.forename: Chenggui
authors.initials: C
authors.lastname: Han
authors.forename: Wan-Xiang
authors.initials: WX
authors.lastname: Li
authors.forename: Shou-Wei
authors.initials: SW
authors.lastname: Ding
mesh_terms: D000818:Animals; D004330:Drosophila / Q000276:immunology* / Q000821:virology*; D059014:High-Throughput Nucleotide Sequencing; D030381:Nodaviridae / Q000235:genetics / Q000276:immunology*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000276:immunology*; D012367:RNA, Viral / Q000235:genetics; D017384:Sequence Deletion; D016019:Survival Analysis
chemical_list: D034741:RNA, Small Interfering; D012367:RNA, Viral