Signatures of Nucleotide Analog Incorporation by an RNA-Dependent RNA Polymerase Revealed Using High-Throughput Magnetic Tweezers

Dulin D, Arnold JJ, Van Laar T, Oh HS, Lee C, Perkins AL, Harki DA, Depken M, Cameron CE, Dekker NH (2017)

Publication Type: Journal article

Publication year: 2017

Journal

Cell Reports Elsevier (Cell Press)

Book Volume: 21

Pages Range: 1063-1076

Journal Issue: 4

DOI: 10.1016/j.celrep.2017.10.005

Abstract

RNA viruses pose a threat to public health that is exacerbated by the dearth of antiviral therapeutics. The RNA-dependent RNA polymerase (RdRp) holds promise as a broad-spectrum, therapeutic target because of the conserved nature of the nucleotide-substrate-binding and catalytic sites. Conventional, quantitative, kinetic analysis of antiviral ribonucleotides monitors one or a few incorporation events. Here, we use a high-throughput magnetic tweezers platform to monitor the elongation dynamics of a prototypical RdRp over thousands of nucleotide-addition cycles in the absence and presence of a suite of nucleotide analog inhibitors. We observe multiple RdRp-RNA elongation complexes; only a subset of which are competent for analog utilization. Incorporation of a pyrazine-carboxamide nucleotide analog, T-1106, leads to RdRp backtracking. This analysis reveals a mechanism of action for this antiviral ribonucleotide that is corroborated by cellular studies. We propose that induced backtracking represents a distinct mechanistic class of antiviral ribonucleotides.

Authors with CRIS profile

David Dulin Interdisziplinäres Zentrum für Klinische Forschung IZKF Nachwuchsgruppe 2 Prof. Dr. Dulin

Involved external institutions

Pennsylvania State University (Penn State) US United States (USA) (US) Delft University of Technology (TU Delft) NL Netherlands (NL) University of Minnesota (UMN) US United States (USA) (US)

How to cite

APA:

Dulin, D., Arnold, J.J., Van Laar, T., Oh, H.-S., Lee, C., Perkins, A.L.,... Dekker, N.H. (2017). Signatures of Nucleotide Analog Incorporation by an RNA-Dependent RNA Polymerase Revealed Using High-Throughput Magnetic Tweezers. Cell Reports, 21(4), 1063-1076. https://dx.doi.org/10.1016/j.celrep.2017.10.005

MLA:

Dulin, David, et al. "Signatures of Nucleotide Analog Incorporation by an RNA-Dependent RNA Polymerase Revealed Using High-Throughput Magnetic Tweezers." Cell Reports 21.4 (2017): 1063-1076.

BibTeX: Download