Serine ADP-ribosylation marks nucleosomes for ALC1-dependent chromatin remodeling
- The University of Texas Southwestern Medical Center, United States
Abstract
Serine ADP-ribosylation (ADPr) is a DNA damage-induced post-translational modification catalyzed by the PARP1/2:HPF1 complex. As the list of PARP1/2:HPF1 substrates continues to expand, there is a need for technologies to prepare mono- and poly-ADP-ribosylated proteins for biochemical interrogation. Here we investigate the unique peptide ADPr activities catalyzed by PARP1 in the absence and presence of HPF1. We then exploit these activities to develop a method that facilitates installation of ADP-ribose polymers onto peptides with precise control over chain length and modification site. Importantly, the enzymatically mono- and poly-ADP-ribosylated peptides are fully compatible with protein ligation technologies. This chemoenzymatic protein synthesis strategy was employed to assemble a series of full-length, ADP-ribosylated histones and show that ADPr at H2BS6 or H3S10 converts nucleosomes into robust substrates for the chromatin remodeler ALC1. We found ALC1 preferentially remodels 'activated' substrates within heterogeneous mononucleosome populations and asymmetrically ADP-ribosylated dinucleosome substrates, and that nucleosome serine ADPr is sufficient to stimulate ALC1 activity in nuclear extracts. Our study identifies a biochemical function for nucleosome serine ADPr and describes a new, highly modular approach to explore the impact that site-specific serine mono- and poly-ADPr have on protein function.
Data availability
All data generated or analysed during this study are included in the manuscript and supporting files. Supplementary Dataset is available online on dryad- https://doi.org/10.5061/dryad.z612jm6cc
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Data from: Serine ADP-ribosylation marks nucleosomes for ALC1-dependent chromatin remodelingDryad Digital Repository, doi:10.5061/dryad.z612jm6cc.
Article and author information
Author details
Funding
Cancer Prevention and Research Institute of Texas (RR180051)
- Glen Liszczak
Welch Foundation (I-2039)
- Glen Liszczak
American Cancer Society (UTSW-IRG-17-174-13)
- Glen Liszczak
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2021, Mohapatra 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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