Serine ADP-ribosylation marks nucleosomes for ALC1-dependent chromatin remodeling.
Serine ADP-ribosylation marks nucleosomes for ALC1-dependent chromatin remodeling.
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DOI：
10.7554/elife.71502
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发表时间：
2021-12-07
期刊：
影响因子：
7.7
通讯作者：
Liszczak G
中科院分区：
文献类型：
--
作者：
Mohapatra J;Tashiro K;Beckner RL;Sierra J;Kilgore JA;Williams NS;Liszczak G
丝氨酸ADP核糖基化（Serine ADP ribosylation，ADPr）是由PARP 1/2：HPF 1复合物催化的DNA损伤诱导的翻译后修饰。随着PARP 1/2：HPF 1底物清单的不断扩大，需要制备用于生物化学询问的单ADP核糖基化蛋白和多ADP核糖基化蛋白的技术。在这里，我们研究了独特的肽ADPr活动催化PARP 1的存在和不存在的HPF 1。然后，我们利用这些活动，以开发一种方法，便于安装ADP-核糖聚合物的肽与精确控制的链长和修饰位点。重要的是，酶促单ADP核糖基化肽和多ADP核糖基化肽与蛋白质连接技术完全相容。这种化学酶促蛋白质合成策略被用来组装一系列全长ADP核糖基化组蛋白，并显示组蛋白H2 B丝氨酸6或组蛋白H3丝氨酸10处的ADPr将核小体转化为染色质重塑剂ALC 1的稳健底物。我们发现，ALC 1优先重塑“激活”的基板内异质monolithome人口和不对称的ADP-核糖基化dinucleosome基板，和核小体丝氨酸ADPr是足以刺激ALC 1活性的核提取物。我们的研究确定了核小体丝氨酸ADPr的生化功能，并描述了一种新的，高度模块化的方法来探索位点特异性丝氨酸单和多聚ADPr对蛋白质功能的影响。
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.