RNA interference functions as an antiviral immunity mechanism in mammals - PubMed (original) (raw)
RNA interference functions as an antiviral immunity mechanism in mammals
Yang Li et al. Science. 2013.
Abstract
Diverse eukaryotic hosts produce virus-derived small interfering RNAs (siRNAs) to direct antiviral immunity by RNA interference (RNAi). However, it remains unknown whether the mammalian RNAi pathway has a natural antiviral function. Here, we show that infection of hamster cells and suckling mice by Nodamura virus (NoV), a mosquito-transmissible RNA virus, requires RNAi suppression by its B2 protein. Loss of B2 expression or its suppressor activity leads to abundant production of viral siRNAs and rapid clearance of the mutant viruses in mice. However, viral small RNAs detected during virulent infection by NoV do not have the properties of canonical siRNAs. These findings have parallels with the induction and suppression of antiviral RNAi by the related Flock house virus in fruit flies and nematodes and reveal a mammalian antiviral immunity mechanism mediated by RNAi.
Figures
Fig. 1. siRNA properties of vsRNAs in BHK-21 cells
(A) Length distribution and abundance of positive- or negative-strand vsRNAs from cells 2 or 3 dpi with NoV or NoVΔB2. (B) Total counts of pairs of complementary 22-nt vsRNAs of NoV or NoVΔB2 in each distance category (in nucleotides) between 5′ and 3′ ends of a complementary vsRNA pair, defined as 0 for perfect base-paired 22-nt vsRNAs with blunt ends, –2 for pairs with 2-nt overhang at the 3′-end of each strand (α and β), or 20 for pairs with 20-nt overhang at the 5′-ends (α and γ).
Fig. 2. NoV infection requires RNAi suppression
(A) BHK-21 cells or BHK cells stably expressing B2 or VP35 were mock-infected or infected by NoVΔB2 or NoV of the same titer. Every 12 hours postinfection (hpi), the viral genomic RNA1 levels were determined by quantitative RT-PCR with the accumulation level of NoVΔB2 in BHK-21 cells at 12 hpi set as 1. Error bars indicate standard deviation of three replicates. (B) Accumulation of NoV and NoVΔB2 RNAs 1 to 3 in the infected cells detected by Northern blotting. RNA1 signal quantified by phosphorimaging was shown with that of NoVΔB2 in BHK-21 cells (lanes 4) set as 1. Detection of B2 transgene mRNA (arrow) was visible. 18_S_ rRNA staining served as loading control.
Fig. 3. In vivo virus clearance associated with production of viral siRNAs
(A and B) Accumulation of NoV, NoVΔB2, and NoVmB2 in mouse fore- (F) and hind- (H) limb tissues detected by quantitative RT-PCR of the viral RNA1 and Northern blotting, respectively. NoVΔB2 level in hind limb at 1 dpi was set as 1, and error bars indicate standard deviation of three replicates (A). NoV RNAs 1 and 2 (arrows) were visible after rRNA staining to show equal loading (B). (C) Suckling mice remained as healthy 4 weeks post-infection with either NoVΔB2 (right) or NoVmB2 (not shown) as mock-inoculated mice (left), whereas all of the five NoV-inoculated mice died by 5 dpi (not shown). (D) Northern blot detection of negative-strand viral siRNAs in mice infected with NoVΔB2 (middle) or NoVmB2 (left) and of vsRNAs from NoV-infected mice (right). The hybridizing positions of four siRNA probes were given in Fig. 4B, and size markers were synthetic 21- and 25-nt RNAs. The same filters were probed for mouse microRNA 127 (miR-127) and U6 RNA as loading controls. At least three independent repeats with reproducible results were performed with each experiment.
Fig. 4. Properties of mouse viral siRNAs produced in vivo
(A) Length distribution and abundance of positive- or negative-strand vsRNAs from mice 1 or 2 dpi with NoVΔB2 or with NoV at 4 dpi. (B) Total counts of pairs of complementary 22-nt vsRNAs of NoVΔB2 and NoV in each distance category as defined in Fig. 1B. (C) Virus genome distribution of 21-to 23-nt viral siRNAs sequenced from either sucking mice (top two panels) or BHK-21 cells (bottom two panels) after infection by NoVΔB2. The functional proteins encoded by the viral bipartite RNA genome and transcription of B2 mRNA (RNA3) from RNA1 are shown. Arrows indicate the positions of the four locked nucleic acid probes used to detect negative-strand viral siRNAs in mice.
Comment in
- Molecular biology. RNAi, Antiviral after all.
Sagan SM, Sarnow P. Sagan SM, et al. Science. 2013 Oct 11;342(6155):207-8. doi: 10.1126/science.1245475. Science. 2013. PMID: 24115433 No abstract available. - Small RNAs: antiviral RNAi in mammals.
Burgess DJ. Burgess DJ. Nat Rev Genet. 2013 Dec;14(12):821. doi: 10.1038/nrg3616. Epub 2013 Oct 22. Nat Rev Genet. 2013. PMID: 24145213 No abstract available. - Antiviral RNA interference in animals: piecing together the evidence.
Tanguy M, Miska EA. Tanguy M, et al. Nat Struct Mol Biol. 2013 Nov;20(11):1239-41. doi: 10.1038/nsmb.2708. Nat Struct Mol Biol. 2013. PMID: 24197164 No abstract available.
Similar articles
- Small RNAs: antiviral RNAi in mammals.
Burgess DJ. Burgess DJ. Nat Rev Genet. 2013 Dec;14(12):821. doi: 10.1038/nrg3616. Epub 2013 Oct 22. Nat Rev Genet. 2013. PMID: 24145213 No abstract available. - Mechanism and Function of Antiviral RNA Interference in Mice.
Han Q, Chen G, Wang J, Jee D, Li WX, Lai EC, Ding SW. Han Q, et al. mBio. 2020 Aug 4;11(4):e03278-19. doi: 10.1128/mBio.03278-19. mBio. 2020. PMID: 32753500 Free PMC article. - Molecular biology. RNAi, Antiviral after all.
Sagan SM, Sarnow P. Sagan SM, et al. Science. 2013 Oct 11;342(6155):207-8. doi: 10.1126/science.1245475. Science. 2013. PMID: 24115433 No abstract available. - RNA-based viral immunity initiated by the Dicer family of host immune receptors.
Aliyari R, Ding SW. Aliyari R, et al. Immunol Rev. 2009 Jan;227(1):176-88. doi: 10.1111/j.1600-065X.2008.00722.x. Immunol Rev. 2009. PMID: 19120484 Free PMC article. Review. - Antiviral RNAi in Insects and Mammals: Parallels and Differences.
Schuster S, Miesen P, van Rij RP. Schuster S, et al. Viruses. 2019 May 16;11(5):448. doi: 10.3390/v11050448. Viruses. 2019. PMID: 31100912 Free PMC article. Review.
Cited by
- RNAi-Induced Gene Silencing against Chikungunya and COVID-19: What Have We Learned So Far, and What Is the Way Forward?
Panda K, Alagarasu K, Tagore R, Paingankar M, Kumar S, Jeengar MK, Cherian S, Parashar D. Panda K, et al. Viruses. 2024 Sep 20;16(9):1489. doi: 10.3390/v16091489. Viruses. 2024. PMID: 39339965 Free PMC article. Review. - The impact of host microRNAs on the development of conserved mutations of SARS-CoV-2.
Ghaemi S, Abdoli A, Karimi H, Saadatpour F, Arefian E. Ghaemi S, et al. Sci Rep. 2024 Sep 27;14(1):22091. doi: 10.1038/s41598-024-70974-7. Sci Rep. 2024. PMID: 39333651 Free PMC article. - Protein-coding circular RNA enhances antiviral immunity via JAK/STAT pathway in Drosophila.
Guo D, Xu W, Cui T, Rong Q, Wu Q. Guo D, et al. mBio. 2024 Sep 11;15(9):e0146924. doi: 10.1128/mbio.01469-24. Epub 2024 Aug 19. mBio. 2024. PMID: 39158293 Free PMC article. - Functional canonical RNAi in mice expressing a truncated Dicer isoform and long dsRNA.
Buccheri V, Pasulka J, Malik R, Loubalova Z, Taborska E, Horvat F, Roos Kulmann MI, Jenickova I, Prochazka J, Sedlacek R, Svoboda P. Buccheri V, et al. EMBO Rep. 2024 Jul;25(7):2896-2913. doi: 10.1038/s44319-024-00148-z. Epub 2024 May 20. EMBO Rep. 2024. PMID: 38769420 Free PMC article. - Live-attenuated virus vaccine defective in RNAi suppression induces rapid protection in neonatal and adult mice lacking mature B and T cells.
Chen G, Han Q, Li WX, Hai R, Ding SW. Chen G, et al. Proc Natl Acad Sci U S A. 2024 Apr 23;121(17):e2321170121. doi: 10.1073/pnas.2321170121. Epub 2024 Apr 17. Proc Natl Acad Sci U S A. 2024. PMID: 38630724 Free PMC article.
References
- Hamilton AJ, Baulcombe DC. Science. 1999;286:950–952. - PubMed
- Li HW, Li WX, Ding SW. Science. 2002;296:1319–1321. - PubMed
- Wilkins C, et al. Nature. 2005;436:1044–1047. - PubMed
Publication types
MeSH terms
Substances
Grants and funding
- RC1 GM091896/GM/NIGMS NIH HHS/United States
- GM94396/GM/NIGMS NIH HHS/United States
- R01 AI052447/AI/NIAID NIH HHS/United States
- AI52447/AI/NIAID NIH HHS/United States
- R01 GM094396/GM/NIGMS NIH HHS/United States
LinkOut - more resources
Full Text Sources
Other Literature Sources