The online version of this article (https://doi.org/10.1186/s12985-017-0915-1) contains supplementary material, which is available to authorized users.
Influenza is a severe contagious disease especially in children, elderly and immunocompromised patients. Beside vaccination, the discovery of new anti-viral agents represents an important strategy to encounter seasonal and pandemic influenza A virus (IAV) strains. The bacterial extra-cellular ribonuclease binase is a well-studied RNase from Bacillus pumilus. Treatment with binase was shown to improve survival of laboratory animals infected with different RNA viruses. Although binase reduced IAV titer in vitro and in vivo, the mode of action (MOA) of binase against IAV at the molecular level has yet not been studied in depth and remains elusive.
To analyze whether binase impairs virus replication by direct interaction with the viral particle we applied a hemagglutination inhibition assay and monitored the integrity of the viral RNA within the virus particle by RT-PCR. Furthermore, we used Western blot and confocal microscopy analysis to study whether binase can internalize into MDCK-II cells. By primer extension we examined the effect of binase on the integrity of viral RNAs within the cells and using a mini-genome system we explored the effect of binase on the viral expression.
We show that (i) binase does not to attack IAV particle-protected viral RNA, (ii) internalized binase could be detected within the cytosol of MDCK-II cells and that (iii) binase impairs IAV replication by specifically degrading viral RNA species within the infected MDCK-II cells without obvious effect on cellular mRNAs.
Our data provide novel evidence suggesting that binase is a potential anti-viral agent with specific intra-cellular MOA.
Additional file 1: Figure S1. The effect of binase on HA inhibition (HI). Twofold serially diluted binase (105 U/ml) was incubated with 4HAU of H1N1pdm09 and then supplemented with 1% chicken erythrocytes (RBCs) to detect the binase/ligand interaction (binase + H1N1pdm + RBCs). Meanwhile, twofold serially diluted binase (105 U/ml) was incubated with 1% chicken erythrocytes (RBCs) and then 4 HAU of H1N1pdm09 were added to explore the binase/receptor interaction (binase + RBCs + H1N1pdm). HI assay was conducted in triplicate for each serial dilution (1:10–1:1280).1xPBS, binase (105 U/ml), and H1N1pdm09 (4 HAU) were used as controls. (PDF 432 kb)12985_2017_915_MOESM1_ESM.pdf
Oh DY, Hurt AC. a review of the antiviral susceptibility of human and avian influenza viruses over the last decade. Scientifica (Cairo). 2014;2014:430629.
Ilinskaya ON, Shah Mahmud R. Ribonucleases as antiviral agents. Mol Biol. 2014;48:615–23. CrossRef
Shah Mahmud R, Ilinskaya ON. Antiviral activity of Binase against the pandemic influenza a (H1N1) virus. Acta Nat. 2013;5:44–51.
Alekseeva II, Kurinenko BM, Kleiner GI, Skuia A, Penzikova GA. Comparative study of the antiviral activity of pancreatic and microbial RNAse. Antibiotiki. 1981;26:527–32. PubMed
Efimova MA, Shah Mahmud R, Zelenikhin PV, Sabirova MI, Kolpakov AI, Ilinskaya ON. Exogenous Bacillus Pumilus RNase (Binase) suppresses the reproduction of Reovirus serotype 1. Mol Biol. 2017;51:96–101. CrossRef
Muller C, Ulyanova V, Ilinskaya O, Pleschka S, Mahmud RS, Novel Antiviral A. Strategy against MERS-CoV and HCoV-229E using Binase to target viral genome replication. Bionanoscience. 2017;7:294–9. CrossRef
Gribencha SV, Potselueva LA, Barinskii IF, Deev SM, Balandin TG, Leshchinskaia IB. Antiviral activity of Bacillus Intermedius RNAase in guinea-pigs and rabbits infected with outdoor rabies virus. Vopr Virusol. 2006;51:41–3. PubMed
Shneider MA, Shtil'bans EB, Kuprianov-Ashin EG, Potselueva LA, Zaikonnikova IV, Kurinenko BM. anti-influenza effect of bacterial RNAse and the pharmacokinetic basis of its administration in experimental studies. Antibiot Khimioter. 1990;35:27–31. PubMed
Shah Mahmud R, Efimova M, Mostafa A, Ulyanova V, Ilinskaya O. Antiviral activity of bacterial extracellular ribonuclease against single-, double-stranded RNA and DNA containing viruses in cell cultures. Bionanoscience. 2016;6:561–3. CrossRef
Nacken W, Anhlan D, Hrincius ER, Mostafa A, Wolff T, Sadewasser A, Pleschka S, Ehrhardt C, Ludwig S. Activation of c-jun N-terminal kinase upon influenza a virus (IAV) infection is independent of pathogen-related receptors but dependent on amino acid sequence variations of IAV NS1. J Virol. 2014;88:8843–52. CrossRefPubMedPubMedCentral
- Bacterial ribonuclease binase exerts an intra-cellular anti-viral mode of action targeting viral RNAs in influenza a virus-infected MDCK-II cells
Raihan Shah Mahmud
- BioMed Central
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