RNF20-mediated H2B monoubiquitination protects stalled forks from degradation and promotes fork restart.

IF 6.5 1区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

EMBO Reports Pub Date : 2025-06-10 DOI:10.1038/s44319-025-00497-3

Debanjali Bhattacharya, Harsh Kumar Dwivedi, Ganesh Nagaraju

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引用次数: 0

Abstract

Chromatin modifications play an important role in transcription, DNA replication and repair. Nonetheless, whether histone modifications regulate replication stress responses remains obscure. Here, we show that RNF20 localizes to and promotes H2B monoubiquitination (H2Bub) at replicating sites. Knockdown of RNF20 leads to degradation of stalled forks by nucleolytic enzymes, which can be rescued by inhibition of MRE11/DNA2 and co-depletion of SMARCAL1/HLTF/ZRANB3 fork remodelers. RNF20 facilitates the loading of RAD51 and RAD51C at stalled fork sites and acts in the same pathway of RAD51/RAD51C-mediated fork protection and restart. Analyses with RING domain and phosphorylation-deficient mutants of RNF20 show that its catalytic activity and ATR-mediated phosphorylation are essential for its role in replication stress responses. Finally, treatment of RNF20-depleted cells with chromatin relaxing agents rescues fork protection and restart defects. Collectively, our study uncovers a role for RNF20-mediated H2Bub in regulating chromatin dynamics to safeguard replicating genomes.

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rnf20介导的H2B单泛素化保护停滞分叉免受降解并促进分叉重启。

染色质修饰在转录、DNA复制和修复中起着重要作用。然而，组蛋白修饰是否调节复制应激反应仍然不清楚。在这里，我们发现RNF20定位并促进复制位点的H2B单泛素化（H2Bub）。RNF20的敲低会导致停顿叉被核溶酶降解，这可以通过抑制MRE11/DNA2和共同耗尽SMARCAL1/HLTF/ZRANB3叉重塑子来挽救。RNF20促进了RAD51和RAD51C在停滞分叉位点的加载，并在RAD51/RAD51C介导的分叉保护和重启的同一途径中起作用。对RNF20的RING结构域和磷酸化缺陷突变体的分析表明，其催化活性和atr介导的磷酸化对其在复制应激反应中的作用至关重要。最后，用染色质松弛剂处理rnf20缺失的细胞可以挽救分叉保护和重启缺陷。总的来说，我们的研究揭示了rnf20介导的H2Bub在调节染色质动力学以保护基因组复制中的作用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

EMBO Reports 生物-生化与分子生物学

CiteScore

11.20

自引率

1.30%

发文量

267

审稿时长

1 months

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