Chemical genetics and proteome-wide site mapping reveal cysteine MARylation by PARP-7 on immune-relevant protein targets
- Oregon Health and Science University, United States
- University of Copenhagen, Denmark
- University of California, San Francisco, United States
- University of Toronto, Canada
- University of Oslo, Norway
Abstract
Poly(ADP-ribose) polymerase 7 (PARP-7) has emerged as a critically important member of a large enzyme family that catalyzes ADP-ribosylation in mammalian cells. PARP-7 is a critical regulator of the innate immune response. What remains unclear is the mechanism by which PARP-7 regulates this process, namely because the protein targets of PARP-7 mono-ADP-ribosylation (MARylation) are largely unknown. Here, we combine chemical genetics, proximity labeling, and proteome-wide amino acid ADP-ribosylation site profiling for identifying the direct targets and sites of PARP-7-mediated MARylation in a cellular context. We found that the inactive PARP family member, PARP-13—a critical regulator of the antiviral innate immune response—is a major target of PARP-7. PARP-13 is preferentially MARylated on cysteine residues in its RNA binding zinc finger domain. Proteome-wide ADP-ribosylation analysis reveals cysteine as a major MARylation acceptor of PARP-7. This study provides insight into PARP-7 targeting and MARylation site preference.
Data availability
All RAW proteomics files have been uploaded to PRIDE. This is related to all Supplementary Tables.
Article and author information
Author details
Sara C Buch-Larsen
Funding
National Institute of Neurological Disorders and Stroke (NIH 2R01NS088629)
- Michael S Cohen
Pew Charitable Trusts (NA)
- Michael S Cohen
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Anthony K. L. Leung
Version history
- Received: June 27, 2020
- Accepted: January 15, 2021
- Accepted Manuscript published: January 21, 2021 (version 1)
- Version of Record published: February 12, 2021 (version 2)
- Version of Record updated: March 2, 2021 (version 3)
Copyright
© 2021, Rodriguez et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Chemical genetics and proteome-wide site mapping reveal cysteine MARylation by PARP-7 on immune-relevant protein targets
eLife 10:e60480.
https://doi.org/10.7554/eLife.60480
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