BIOMAT Database Logo BIOMAT Database Logo

Interfering with nucleotide excision by the coronavirus 3'-to-5' exoribonuclease. 2022

Rukesh Chinthapatla, and Mohamad Sotoudegan, and Thomas Anderson, and Ibrahim M Moustafa, and Kellan T Passow, and Samantha A Kennelly, and Ramkumar Moorthy, and David Dulin, and Joy Y Feng, and Daniel A Harki, and Robert Kirchdoerfer, and Craig E Cameron, and Jamie J Arnold
Department of Microbiology and Immunology, The University of North Carolina at Chapel Hill School of Medicine, Chapel Hill, NC 27599, USA.

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.

UI MeSH Term Description Entries

Related Publications

Rukesh Chinthapatla, and Mohamad Sotoudegan, and Thomas Anderson, and Ibrahim M Moustafa, and Kellan T Passow, and Samantha A Kennelly, and Ramkumar Moorthy, and David Dulin, and Joy Y Feng, and Daniel A Harki, and Robert Kirchdoerfer, and Craig E Cameron, and Jamie J Arnold
RNA 3'-end mismatch excision by the severe acute respiratory syndrome coronavirus nonstructural protein nsp10/nsp14 exoribonuclease complex.
June 2012, Proceedings of the National Academy of Sciences of the United States of America,
Rukesh Chinthapatla, and Mohamad Sotoudegan, and Thomas Anderson, and Ibrahim M Moustafa, and Kellan T Passow, and Samantha A Kennelly, and Ramkumar Moorthy, and David Dulin, and Joy Y Feng, and Daniel A Harki, and Robert Kirchdoerfer, and Craig E Cameron, and Jamie J Arnold
Drosophila 5'-->3'-exoribonuclease Pacman.
January 2001, Methods in enzymology,
Rukesh Chinthapatla, and Mohamad Sotoudegan, and Thomas Anderson, and Ibrahim M Moustafa, and Kellan T Passow, and Samantha A Kennelly, and Ramkumar Moorthy, and David Dulin, and Joy Y Feng, and Daniel A Harki, and Robert Kirchdoerfer, and Craig E Cameron, and Jamie J Arnold
5'-->3'-exoribonuclease from rabbit reticulocytes.
January 2001, Methods in enzymology,
Rukesh Chinthapatla, and Mohamad Sotoudegan, and Thomas Anderson, and Ibrahim M Moustafa, and Kellan T Passow, and Samantha A Kennelly, and Ramkumar Moorthy, and David Dulin, and Joy Y Feng, and Daniel A Harki, and Robert Kirchdoerfer, and Craig E Cameron, and Jamie J Arnold
In vitro assays of 5' to 3'-exoribonuclease activity.
January 2008, Methods in enzymology,
Rukesh Chinthapatla, and Mohamad Sotoudegan, and Thomas Anderson, and Ibrahim M Moustafa, and Kellan T Passow, and Samantha A Kennelly, and Ramkumar Moorthy, and David Dulin, and Joy Y Feng, and Daniel A Harki, and Robert Kirchdoerfer, and Craig E Cameron, and Jamie J Arnold
Purification of a human polyribosome-associated 3' to 5' exoribonuclease.
December 1994, The Journal of biological chemistry,
Rukesh Chinthapatla, and Mohamad Sotoudegan, and Thomas Anderson, and Ibrahim M Moustafa, and Kellan T Passow, and Samantha A Kennelly, and Ramkumar Moorthy, and David Dulin, and Joy Y Feng, and Daniel A Harki, and Robert Kirchdoerfer, and Craig E Cameron, and Jamie J Arnold
XRN1: A Major 5' to 3' Exoribonuclease in Eukaryotic Cells.
January 2012, The Enzymes,
Rukesh Chinthapatla, and Mohamad Sotoudegan, and Thomas Anderson, and Ibrahim M Moustafa, and Kellan T Passow, and Samantha A Kennelly, and Ramkumar Moorthy, and David Dulin, and Joy Y Feng, and Daniel A Harki, and Robert Kirchdoerfer, and Craig E Cameron, and Jamie J Arnold
In vivo 3'-to-5' exoribonuclease targetomes of Streptococcus pyogenes.
November 2018, Proceedings of the National Academy of Sciences of the United States of America,
Rukesh Chinthapatla, and Mohamad Sotoudegan, and Thomas Anderson, and Ibrahim M Moustafa, and Kellan T Passow, and Samantha A Kennelly, and Ramkumar Moorthy, and David Dulin, and Joy Y Feng, and Daniel A Harki, and Robert Kirchdoerfer, and Craig E Cameron, and Jamie J Arnold
Discovery of an RNA virus 3'->5' exoribonuclease that is critically involved in coronavirus RNA synthesis.
March 2006, Proceedings of the National Academy of Sciences of the United States of America,
Rukesh Chinthapatla, and Mohamad Sotoudegan, and Thomas Anderson, and Ibrahim M Moustafa, and Kellan T Passow, and Samantha A Kennelly, and Ramkumar Moorthy, and David Dulin, and Joy Y Feng, and Daniel A Harki, and Robert Kirchdoerfer, and Craig E Cameron, and Jamie J Arnold
The 3'-to-5' exoribonuclease Nibbler shapes the 3' ends of microRNAs bound to Drosophila Argonaute1.
November 2011, Current biology : CB,
Rukesh Chinthapatla, and Mohamad Sotoudegan, and Thomas Anderson, and Ibrahim M Moustafa, and Kellan T Passow, and Samantha A Kennelly, and Ramkumar Moorthy, and David Dulin, and Joy Y Feng, and Daniel A Harki, and Robert Kirchdoerfer, and Craig E Cameron, and Jamie J Arnold
Biochemical characterization of exoribonuclease encoded by SARS coronavirus.
September 2007, Journal of biochemistry and molecular biology,
Copied contents to your clipboard!