tetano
Editor, Senior Moderator
J Virol
. 2020 Nov 6;JVI.01969-20.
doi: 10.1128/JVI.01969-20. Online ahead of print.
The SARS-CoV-2 conserved macrodomain is a mono-ADP-ribosylhydrolase
Yousef M O Alhammad[SUP] 1 [/SUP], Maithri M Kashipathy[SUP] 2 [/SUP], Anuradha Roy[SUP] 3 [/SUP], Jean-Philippe Gagn?[SUP] 4 5 [/SUP], Peter McDonald[SUP] 3 [/SUP], Philip Gao[SUP] 6 [/SUP], Louis Nonfoux[SUP] 4 5 [/SUP], Kevin P Battaile[SUP] 7 [/SUP], David K Johnson[SUP] 8 [/SUP], Erik D Holmstrom[SUP] 1 [/SUP], Guy G Poirier[SUP] 4 5 [/SUP], Scott Lovell[SUP] 2 [/SUP], Anthony R Fehr[SUP] 9 [/SUP]
Affiliations
Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and other SARS-related CoVs encode 3 tandem macrodomains within non-structural protein 3 (nsp3). The first macrodomain, Mac1, is conserved throughout CoVs, and binds to and hydrolyzes mono-ADP-ribose (MAR) from target proteins. Mac1 likely counters host-mediated anti-viral ADP-ribosylation, a posttranslational modification that is part of the host response to viral infections. Mac1 is essential for pathogenesis in multiple animal models of CoV infection, implicating it as a virulence factor and potential therapeutic target. Here we report the crystal structure of SARS-CoV-2 Mac1 in complex with ADP-ribose. SARS-CoV-2, SARS-CoV and MERS-CoV Mac1 exhibit similar structural folds and all 3 proteins bound to ADP-ribose with low μM affinities. Importantly, using ADP-ribose detecting binding reagents in both a gel-based assay and novel ELISA assays, we demonstrated de-MARylating activity for all 3 CoV Mac1 proteins, with the SARS-CoV-2 Mac1 protein leading to a more rapid loss of substrate compared to the others. In addition, none of these enzymes could hydrolyze poly-ADP-ribose. We conclude that the SARS-CoV-2 and other CoV Mac1 proteins are MAR-hydrolases with similar functions, indicating that compounds targeting CoV Mac1 proteins may have broad anti-CoV activity.IMPORTANCE SARS-CoV-2 has recently emerged into the human population and has led to a worldwide pandemic of COVID-19 that has caused greater than 1.2 million deaths worldwide. With, no currently approved treatments, novel therapeutic strategies are desperately needed. All coronaviruses encode for a highly conserved macrodomain (Mac1) that binds to and removes ADP-ribose adducts from proteins in a dynamic post-translational process increasingly recognized as an important factor that regulates viral infection. The macrodomain is essential for CoV pathogenesis and may be a novel therapeutic target. Thus, understanding its biochemistry and enzyme activity are critical first steps for these efforts. Here we report the crystal structure of SARS-CoV-2 Mac1 in complex with ADP-ribose, and describe its ADP-ribose binding and hydrolysis activities in direct comparison to SARS-CoV and MERS-CoV Mac1 proteins. These results are an important first step for the design and testing of potential therapies targeting this unique protein domain.
. 2020 Nov 6;JVI.01969-20.
doi: 10.1128/JVI.01969-20. Online ahead of print.
The SARS-CoV-2 conserved macrodomain is a mono-ADP-ribosylhydrolase
Yousef M O Alhammad[SUP] 1 [/SUP], Maithri M Kashipathy[SUP] 2 [/SUP], Anuradha Roy[SUP] 3 [/SUP], Jean-Philippe Gagn?[SUP] 4 5 [/SUP], Peter McDonald[SUP] 3 [/SUP], Philip Gao[SUP] 6 [/SUP], Louis Nonfoux[SUP] 4 5 [/SUP], Kevin P Battaile[SUP] 7 [/SUP], David K Johnson[SUP] 8 [/SUP], Erik D Holmstrom[SUP] 1 [/SUP], Guy G Poirier[SUP] 4 5 [/SUP], Scott Lovell[SUP] 2 [/SUP], Anthony R Fehr[SUP] 9 [/SUP]
Affiliations
- PMID: 33158944
- DOI: 10.1128/JVI.01969-20
Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and other SARS-related CoVs encode 3 tandem macrodomains within non-structural protein 3 (nsp3). The first macrodomain, Mac1, is conserved throughout CoVs, and binds to and hydrolyzes mono-ADP-ribose (MAR) from target proteins. Mac1 likely counters host-mediated anti-viral ADP-ribosylation, a posttranslational modification that is part of the host response to viral infections. Mac1 is essential for pathogenesis in multiple animal models of CoV infection, implicating it as a virulence factor and potential therapeutic target. Here we report the crystal structure of SARS-CoV-2 Mac1 in complex with ADP-ribose. SARS-CoV-2, SARS-CoV and MERS-CoV Mac1 exhibit similar structural folds and all 3 proteins bound to ADP-ribose with low μM affinities. Importantly, using ADP-ribose detecting binding reagents in both a gel-based assay and novel ELISA assays, we demonstrated de-MARylating activity for all 3 CoV Mac1 proteins, with the SARS-CoV-2 Mac1 protein leading to a more rapid loss of substrate compared to the others. In addition, none of these enzymes could hydrolyze poly-ADP-ribose. We conclude that the SARS-CoV-2 and other CoV Mac1 proteins are MAR-hydrolases with similar functions, indicating that compounds targeting CoV Mac1 proteins may have broad anti-CoV activity.IMPORTANCE SARS-CoV-2 has recently emerged into the human population and has led to a worldwide pandemic of COVID-19 that has caused greater than 1.2 million deaths worldwide. With, no currently approved treatments, novel therapeutic strategies are desperately needed. All coronaviruses encode for a highly conserved macrodomain (Mac1) that binds to and removes ADP-ribose adducts from proteins in a dynamic post-translational process increasingly recognized as an important factor that regulates viral infection. The macrodomain is essential for CoV pathogenesis and may be a novel therapeutic target. Thus, understanding its biochemistry and enzyme activity are critical first steps for these efforts. Here we report the crystal structure of SARS-CoV-2 Mac1 in complex with ADP-ribose, and describe its ADP-ribose binding and hydrolysis activities in direct comparison to SARS-CoV and MERS-CoV Mac1 proteins. These results are an important first step for the design and testing of potential therapies targeting this unique protein domain.