Bacterial ribonuclease binase exerts an intra-cellular anti-viral mode of action targeting viral RNAs in influenza a virus-infected MDCK-II cells
Abstract Background 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 e...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2018-01-01
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| Series: | Virology Journal |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12985-017-0915-1 |
| _version_ | 1818966740494712832 |
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| author | Raihan Shah Mahmud Ahmed Mostafa Christin Müller Pumaree Kanrai Vera Ulyanova Yulia Sokurenko Julia Dzieciolowski Irina Kuznetsova Olga Ilinskaya Stephan Pleschka |
| author_facet | Raihan Shah Mahmud Ahmed Mostafa Christin Müller Pumaree Kanrai Vera Ulyanova Yulia Sokurenko Julia Dzieciolowski Irina Kuznetsova Olga Ilinskaya Stephan Pleschka |
| author_sort | Raihan Shah Mahmud |
| collection | DOAJ |
| description | Abstract Background 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. Methods 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. Results 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. Conclusion Our data provide novel evidence suggesting that binase is a potential anti-viral agent with specific intra-cellular MOA. |
| first_indexed | 2024-12-20T13:37:42Z |
| format | Article |
| id | doaj.art-0655f220ddf54d24816fc8570ff07865 |
| institution | Directory Open Access Journal |
| issn | 1743-422X |
| language | English |
| last_indexed | 2024-12-20T13:37:42Z |
| publishDate | 2018-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Virology Journal |
| spelling | doaj.art-0655f220ddf54d24816fc8570ff078652022-12-21T19:38:53ZengBMCVirology Journal1743-422X2018-01-0115111210.1186/s12985-017-0915-1Bacterial ribonuclease binase exerts an intra-cellular anti-viral mode of action targeting viral RNAs in influenza a virus-infected MDCK-II cellsRaihan Shah Mahmud0Ahmed Mostafa1Christin Müller2Pumaree Kanrai3Vera Ulyanova4Yulia Sokurenko5Julia Dzieciolowski6Irina Kuznetsova7Olga Ilinskaya8Stephan Pleschka9Institute of Fundamental Medicine and Biology, Kazan Federal UniversityInstitute of Medical Virology, Justus Liebig UniversityInstitute of Medical Virology, Justus Liebig UniversityInstitute of Medical Virology, Justus Liebig UniversityInstitute of Fundamental Medicine and Biology, Kazan Federal UniversityInstitute of Fundamental Medicine and Biology, Kazan Federal UniversityInstitute of Medical Virology, Justus Liebig UniversityInstitute of Medical Virology, Justus Liebig UniversityInstitute of Fundamental Medicine and Biology, Kazan Federal UniversityInstitute of Medical Virology, Justus Liebig UniversityAbstract Background 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. Methods 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. Results 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. Conclusion Our data provide novel evidence suggesting that binase is a potential anti-viral agent with specific intra-cellular MOA.http://link.springer.com/article/10.1186/s12985-017-0915-1RibonucleaseRNaseBinaseInfluenza virusAnti-viral activity |
| spellingShingle | Raihan Shah Mahmud Ahmed Mostafa Christin Müller Pumaree Kanrai Vera Ulyanova Yulia Sokurenko Julia Dzieciolowski Irina Kuznetsova Olga Ilinskaya Stephan Pleschka Bacterial ribonuclease binase exerts an intra-cellular anti-viral mode of action targeting viral RNAs in influenza a virus-infected MDCK-II cells Virology Journal Ribonuclease RNase Binase Influenza virus Anti-viral activity |
| title | Bacterial ribonuclease binase exerts an intra-cellular anti-viral mode of action targeting viral RNAs in influenza a virus-infected MDCK-II cells |
| title_full | Bacterial ribonuclease binase exerts an intra-cellular anti-viral mode of action targeting viral RNAs in influenza a virus-infected MDCK-II cells |
| title_fullStr | Bacterial ribonuclease binase exerts an intra-cellular anti-viral mode of action targeting viral RNAs in influenza a virus-infected MDCK-II cells |
| title_full_unstemmed | Bacterial ribonuclease binase exerts an intra-cellular anti-viral mode of action targeting viral RNAs in influenza a virus-infected MDCK-II cells |
| title_short | Bacterial ribonuclease binase exerts an intra-cellular anti-viral mode of action targeting viral RNAs in influenza a virus-infected MDCK-II cells |
| title_sort | bacterial ribonuclease binase exerts an intra cellular anti viral mode of action targeting viral rnas in influenza a virus infected mdck ii cells |
| topic | Ribonuclease RNase Binase Influenza virus Anti-viral activity |
| url | http://link.springer.com/article/10.1186/s12985-017-0915-1 |
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