University of Oulu

Lilian Cristina Russo, Rebeka Tomasin, Isaac Araújo Matos, Antonio Carlos Manucci, Sven T. Sowa, Katie Dale, Keith W. Caldecott, Lari Lehtiö, Deborah Schechtman, Flavia C. Meotti, Alexandre Bruni-Cardoso, Nicolas Carlos Hoch, The SARS-CoV-2 Nsp3 macrodomain reverses PARP9/DTX3L-dependent ADP-ribosylation induced by interferon signaling, Journal of Biological Chemistry, Volume 297, Issue 3, 2021, 101041, ISSN 0021-9258,

The SARS-CoV-2 Nsp3 macrodomain reverses PARP9/DTX3L-dependent ADP-ribosylation induced by interferon signaling

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Author: Russo, Lilian Cristina1; Tomasin, Rebeka1; Matos, Isaac Araújo1;
Organizations: 1Department of Biochemistry, Institute of Chemistry, University of São Paulo, São Paulo, Brazil
2Faculty of Biochemistry and Molecular Medicine & Biocenter Oulu, University of Oulu, Oulu, Finland
3Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Falmer, Brighton, United Kingdom
Format: article
Version: published version
Access: open
Online Access: PDF Full Text (PDF, 3.4 MB)
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Language: English
Published: American Society for Biochemistry and Molecular Biology, 2021
Publish Date: 2021-11-24


SARS-CoV-2 nonstructural protein 3 (Nsp3) contains a macrodomain that is essential for coronavirus pathogenesis and is thus an attractive target for drug development. This macrodomain is thought to counteract the host interferon (IFN) response, an important antiviral signalling cascade, via the reversal of protein ADP-ribosylation, a posttranslational modification catalyzed by host poly(ADP-ribose) polymerases (PARPs). However, the main cellular targets of the coronavirus macrodomain that mediate this effect are currently unknown. Here, we use a robust immunofluorescence-based assay to show that activation of the IFN response induces ADP-ribosylation of host proteins and that ectopic expression of the SARS-CoV-2 Nsp3 macrodomain reverses this modification in human cells. We further demonstrate that this assay can be used to screen for on-target and cell-active macrodomain inhibitors. This IFN-induced ADP-ribosylation is dependent on PARP9 and its binding partner DTX3L, but surprisingly the expression of the Nsp3 macrodomain or the deletion of either PARP9 or DTX3L does not impair IFN signaling or the induction of IFN-responsive genes. Our results suggest that PARP9/DTX3L-dependent ADP-ribosylation is a downstream effector of the host IFN response and that the cellular function of the SARS-CoV-2 Nsp3 macrodomain is to hydrolyze this end product of IFN signaling, rather than to suppress the IFN response itself.

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Series: Journal of biological chemistry
ISSN: 0021-9258
ISSN-E: 1083-351X
ISSN-L: 0021-9258
Volume: 297
Issue: 3
Article number: 101041
DOI: 10.1016/j.jbc.2021.101041
Type of Publication: A1 Journal article – refereed
Field of Science: 1182 Biochemistry, cell and molecular biology
3111 Biomedicine
Funding: Work in the N. C. H. lab is supported by a COVID19 emergency grant (2020/53017-6) and a Young Investigator Award (2018/18007-5) from FAPESP, L. C. R. is funded by a FAPESP post-doctoral fellowship and held a Dimensions Sciences short-term fellowship, R. T. is funded by a PNPD/CAPES postdoctoral fellowship, A. C. M. and I. A. M. hold PhD scholarships from CNPq, the F. C. M. lab is supported by a FAPESP Young Investigator II Award (2018/14898-2), and work in the A. B.-C. lab is funded by FAPESP grant 2019/26767-2. Work in the L. L. lab was funded by the Jane and Aatos Erkko Foundation and by the Academy of Finland (grant nos. 287063, 294085, and 319299 to L. L.).
Academy of Finland Grant Number: 287063
Detailed Information: 287063 (Academy of Finland Funding decision)
294085 (Academy of Finland Funding decision)
319299 (Academy of Finland Funding decision)
Copyright information: © 2021 THE AUTHORS. Published by Elsevier Inc on behalf of American Society for Biochemistry and Molecular Biology. This is an open access article under the CC BY license (