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Fidelity Variants of RNA Dependent RNA Polymerases Uncover an Indirect, Mutagenic Activity of Amiloride Compounds

Figure 3

A372V resistance to amiloride does not involve improved replication.

(A) A372V is more resistant than wild type in increasing concentrations of amiloride. HeLa cells were treated with different concentrations of amiloride and infected with wild type or A372V virus at an MOI of 0.01. At 48 h, viable progeny virus was quantified by TCID50 assay. The mean virus titers (TCID50/ml) and S.E.M are shown, N = 3, * P<0.05. (B) One-step growth kinetics of wild type and A372V viruses. HeLa cells were infected at MOI of 10 and the progeny virus was quantified at different hours after infection by TCID50 assay. Mean titers (TCID50/ml) ± S.E.M are shown, N = 3, no significant difference found. (C) The relative increase in signal from time 0 to indicated time points is shown, values are means of 2 separate northen blots of samples from (B), error bars show range of values (D) One-step growth kinetics of wild type and A372V viruses in the presence of 400 µM amiloride. HeLa cells were infected at MOI of 10 and the progeny virus was quantified at different hours after infection by TCID50 assay. Mean titers (TCID50/ml) ± S.E.M are shown, N = 3, no significant differences found except *, p = 0.03. (E) Northern blot analysis of RNA synthesis determined 48 hours after infection with wild type or A372V virus at MOI of 0.01 in the presence or the absence of 400 µM amiloride. Two independent treatment samples per virus are shown.

Figure 3

doi: https://doi.org/10.1371/journal.ppat.1001163.g003