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

PLoS Pathogens

Jamie J. Arnold¹  Bruno Baron²  Malia J. McPherson³  Nina F. Gnädig³  Stéphanie Beaucourt³  Laura I. Levi³  Marco Vignuzzi³ 

[1] Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania, United States of America;Institut Pasteur, Plate Forme de Biophysique des Macromolécules et de leurs Interactions, Paris, France;Institut Pasteur, Viral Populations and Pathogenesis Lab and CNRS URA3015, Paris, France

关键词: Microbial mutation;    RNA synthesis;    Mutagens;    RNA viruses;    Poliovirus;    HeLa cells;    Antimicrobial resistance;    Viral replication;   

DOI  :  10.1371/journal.ppat.1001163

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

In a screen for RNA mutagen resistance, we isolated a high fidelity RNA dependent RNA polymerase (RdRp) variant of Coxsackie virus B3 (CVB3). Curiously, this variant A372V is also resistant to amiloride. We hypothesize that amiloride has a previously undescribed mutagenic activity. Indeed, amiloride compounds increase the mutation frequencies of CVB3 and poliovirus and high fidelity variants of both viruses are more resistant to this effect. We hypothesize that this mutagenic activity is mediated through alterations in intracellular ions such as Mg2+ and Mn2+, which in turn increase virus mutation frequency by affecting RdRp fidelity. Furthermore, we show that another amiloride-resistant RdRp variant, S299T, is completely resistant to this mutagenic activity and unaffected by changes in ion concentrations. We show that RdRp variants resist the mutagenic activity of amiloride via two different mechanisms: 1) increased fidelity that generates virus populations presenting lower basal mutation frequencies or 2) resisting changes in divalent cation concentrations that affect polymerase fidelity. Our results uncover a new antiviral approach based on mutagenesis.

【 授权许可】

CC BY