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Data from: Serine ADP-ribosylation marks nucleosomes for ALC1-dependent chromatin remodeling

Citation

Mohapatra, Jugal; Liszczak, Glen (2022), Data from: Serine ADP-ribosylation marks nucleosomes for ALC1-dependent chromatin remodeling, Dryad, Dataset, https://doi.org/10.5061/dryad.z612jm6cc

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.

Methods

This spreadsheet contains raw data in triplicate for all quantitative experiments associated with the manuscript 'Serine ADP-ribosylation marks nucleosomes for ALC1-dependent chromatin remodeling'. Experiements include HPLC peak integration, fluorescence polarization, densitometry, and quantitative PCR.

Funding

Welch Foundation, Award: I-2039

Cancer Prevention and Research Institute of Texas, Award: RR180051

American Cancer Society, Award: UTSW-IRG-17-174-13