Interfering with nucleotide excision by the coronavirus 3 '-to-5 ' exoribonuclease

Chinthapatla R, Sotoudegan M, Srivastava P, Anderson TK, Moustafa IM, Passow KT, Kennelly SA, Moorthy R, Dulin D, Feng JY, Harki DA, Kirchdoerfer RN, Cameron CE, Arnold JJ (2022)

Publication Type: Journal article

Publication year: 2022

Journal

Nucleic Acids Research Oxford University Press

DOI: 10.1093/nar/gkac1177

Abstract

Some of the most efficacious antiviral therapeutics are ribonucleos(t)ide analogs. The presence of a 3 '-to-5 ' proofreading exoribonuclease (ExoN) in coronaviruses diminishes the potency of many ribonucleotide analogs. The ability to interfere with ExoN activity will create new possibilities for control of SARS-CoV-2 infection. ExoN is formed by a 1:1 complex of nsp14 and nsp10 proteins. We have purified and characterized ExoN using a robust, quantitative system that reveals determinants of specificity and efficiency of hydrolysis. Double-stranded RNA is preferred over single-stranded RNA. Nucleotide excision is distributive, with only one or two nucleotides hydrolyzed in a single binding event. The composition of the terminal basepair modulates excision. A stalled SARS-CoV-2 replicase in complex with either correctly or incorrectly terminated products prevents excision, suggesting that a mispaired end is insufficient to displace the replicase. Finally, we have discovered several modifications to the 3 '-RNA terminus that interfere with or block ExoN-catalyzed excision. While a 3 '-OH facilitates hydrolysis of a nucleotide with a normal ribose configuration, this substituent is not required for a nucleotide with a planar ribose configuration such as that present in the antiviral nucleotide produced by viperin. Design of ExoN-resistant, antiviral ribonucleotides should be feasible.

Authors with CRIS profile

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

Involved external institutions

University of North Carolina at Chapel Hill US United States (USA) (US) University of Minnesota (UMN) US United States (USA) (US) Pennsylvania State University (Penn State) US United States (USA) (US) University of Wisconsin - Madison US United States (USA) (US) Gilead Sciences, Inc. US United States (USA) (US)

How to cite

APA:

Chinthapatla, R., Sotoudegan, M., Srivastava, P., Anderson, T.K., Moustafa, I.M., Passow, K.T.,... Arnold, J.J. (2022). Interfering with nucleotide excision by the coronavirus 3 '-to-5 ' exoribonuclease. Nucleic Acids Research. https://doi.org/10.1093/nar/gkac1177

MLA:

Chinthapatla, Rukesh, et al. "Interfering with nucleotide excision by the coronavirus 3 '-to-5 ' exoribonuclease." Nucleic Acids Research (2022).

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