Serine ADP-ribosylation marks nucleosomes for ALC1-dependent chromatin remodeling

eLife

Kyuto Tashiro¹  Jorge Sierra¹  Glen Liszczak¹  Ryan L Beckner¹  Jugal Mohapatra¹  Jessica A Kilgore²  Noelle S Williams² 

[1] Department of Biochemistry, The University of Texas Southwestern Medical Center, Dallas, United States;Department of Biochemistry, The University of Texas Southwestern Medical Center, Dallas, United States;Preclinical Pharmacology Core, Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, United States;

关键词: DNA damage response;    post-translational modifications;    chemical biology;    protein semi-synthesis;    PARP;    HPF1;    E. coli;    Human;   

DOI  :  10.7554/eLife.71502

来源: eLife Sciences Publications, Ltd

【 摘 要 】

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 histone H2B serine 6 or histone H3 serine 10 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.

【 授权许可】

CC BY