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

Author(s):Show less -

Russo, Lilian Cristina ^[1] ; Tomasin, Rebeka ^[1] ; Matos, Isaac Araujo ^[1] ; Manucci, Antonio Carlos ^[1] ; Sowa, Sven T. ^{[2, 3]} ; Dale, Katie ^{[4, 5]} ; Caldecott, Keith W. ^[4] ; Lehtio, Lari ^{[2, 3]} ; Schechtman, Deborah ^[1] ; Meotti, Flavia C. ^[1] ; Bruni-Cardoso, Alexandre ^[1] ; Hoch, Nicolas Carlos ^[1]

Affiliation

^[1] Univ Sao Paulo, Inst Chem, Dept Biochem, Sao Paulo - Brazil

^[2] Univ Oulu, Fac Biochem & Mol Med, Oulu - Finland

^[3] Univ Oulu, Bioctr Oulu, Oulu - Finland

^[4] Univ Sussex, Genome Damage & Stabil Ctr, Sch Life Sci, Brighton, E Sussex - England

^[5] UCL, UCL Canc Inst, Dept Oncol, London - England

• Document type: Journal article
• Source: Journal of Biological Chemistry; v. 297, n. 3 SEP 2021.
• Web of Science Citations: 0

Abstract

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/DTX3Ldependent 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. (AU)

• FAPESP's process: 18/18007-5 - Protein ADP-ribosylation: DNA damage signalling and impacts on human health
• Grantee: Nicolas Carlos Hoch
• Support Opportunities: Research Grants - Young Investigators Grants
• FAPESP's process: 18/14898-2 - Investigations of the redox processes in inflammatory response and associated pathologies
• Grantee: Flavia Carla Meotti
• Support Opportunities: Research Grants - Young Investigators Grants - Phase 2
• FAPESP's process: 19/26767-2 - Proteostasis regulation of MST2 in the Hippo pathway
• Grantee: Alexandre Bruni Cardoso
• Support Opportunities: Regular Research Grants