Strategies of Tick-Borne Encephalitis Virus to Escape from Nucleoside Analogue-Mediated Inhibition

Kód výsledku v IS VaVaI

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Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Strategies of Tick-Borne Encephalitis Virus to Escape from Nucleoside Analogue-Mediated Inhibition

Popis výsledku v původním jazyce

31 st ICAR, Porto, Portugal, 11.-15. 6. 2018 - lecture. Tick-borne encephalitis virus (TBEV) replication was observed to be strongly inhibited by 2´-C-methyl or 4´-azido modified nucleosides, as well as by an imino-C-nucleoside BCX-4430. TBEV mutants resistant to the aforementioned nucleoside inhibitors were selected by a serial passaging of TBEV in porcine kidney stable cells in the presence of increasing concentrations of the appropriate compound. The resistance of TBEV to 2´-C-methylated nucleosides was associated with a signature mutation S603T within the active site of the viral RdRp. All-atom molecular dynamics simulations revealed that the S603T RdRp mutant repels a water molecule that coordinates the position of a metal ion cofactor as 2´-C-methylated nucleosides approach the active site. Interestingly, the biological properties of the TBEV mutant viruses were dramatically affected, which was manifested by resistance-associated loss of viral replication efficacy in vitro and a highly attenuated virulence phenotype in mice. TBEV resistantance to BCX-4430 was associated with two amino acid substitutions, E460D and Y453H. A reverse genetics approach (infectious-subgenomic-amplicons-based strategy) revealed, that only the mutation E460D, located within the RdRp active site, was required for a high-level resistance to BCX-4430. A molecular docking showed that the mutant D460 provides substantially limited hydrogen bonding interactions with BCX-4430 compared with the E460 wild type, which could explain the resistance mechanism of the E460D mutant to BCX-4430 nucleoside. Surprisingly, the subsitution Y453H was found also in TBEV mutants selected under the pressure of 4´-C-azidocytidine, however, the effect of Y453H on TBEV resistance to 4´-azido modified nucleosides remains to be unclear.

Název v anglickém jazyce

Strategies of Tick-Borne Encephalitis Virus to Escape from Nucleoside Analogue-Mediated Inhibition

Popis výsledku anglicky

31 st ICAR, Porto, Portugal, 11.-15. 6. 2018 - lecture. Tick-borne encephalitis virus (TBEV) replication was observed to be strongly inhibited by 2´-C-methyl or 4´-azido modified nucleosides, as well as by an imino-C-nucleoside BCX-4430. TBEV mutants resistant to the aforementioned nucleoside inhibitors were selected by a serial passaging of TBEV in porcine kidney stable cells in the presence of increasing concentrations of the appropriate compound. The resistance of TBEV to 2´-C-methylated nucleosides was associated with a signature mutation S603T within the active site of the viral RdRp. All-atom molecular dynamics simulations revealed that the S603T RdRp mutant repels a water molecule that coordinates the position of a metal ion cofactor as 2´-C-methylated nucleosides approach the active site. Interestingly, the biological properties of the TBEV mutant viruses were dramatically affected, which was manifested by resistance-associated loss of viral replication efficacy in vitro and a highly attenuated virulence phenotype in mice. TBEV resistantance to BCX-4430 was associated with two amino acid substitutions, E460D and Y453H. A reverse genetics approach (infectious-subgenomic-amplicons-based strategy) revealed, that only the mutation E460D, located within the RdRp active site, was required for a high-level resistance to BCX-4430. A molecular docking showed that the mutant D460 provides substantially limited hydrogen bonding interactions with BCX-4430 compared with the E460 wild type, which could explain the resistance mechanism of the E460D mutant to BCX-4430 nucleoside. Surprisingly, the subsitution Y453H was found also in TBEV mutants selected under the pressure of 4´-C-azidocytidine, however, the effect of Y453H on TBEV resistance to 4´-azido modified nucleosides remains to be unclear.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

10607 - Virology

Projekt

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Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2018

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů