SPRTN-dependent DPC degradation precedes repair of damaged DNA: a proof of concept revealed by the STAR assay

被引：7

作者：

Glumac, Mateo ^{[1
]}

Polovic, Mirjana ^{[1
]}

Batel, Anja ^{[1
]}

Gelemanovic, Andrea ^{[2
]}

Macek, Boris ^{[3
]}

Velic, Ana ^{[3
]}

Marinovic-Terzic, Ivana ^{[1
]}

机构：

[1] Univ Split, Sch Med, Lab Canc Res, Split 21000, Croatia

[2] Mediterranean Inst Life Sci MedILS, Split 21000, Croatia

[3] Univ Tubingen, Interfac Inst Cell Biol, Fac Sci, Quant Proteom, D-72076 Tubingen, Germany

来源：

NUCLEIC ACIDS RESEARCH | 2023年 / 51卷 / 06期

关键词：

PROTEIN CROSS-LINKS; NUCLEOTIDE EXCISION-REPAIR; DOUBLE-STRAND BREAKS; HOMOLOGOUS RECOMBINATION; H2AX PHOSPHORYLATION; HISTONE H2AX; REPLICATION; IRRADIATION; TRINITY; IMAGE;

D O I：

10.1093/nar/gkad022

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

DNA-protein crosslinks (DPCs), formed by the covalent conjugation of proteins to DNA, are toxic lesions that interfere with DNA metabolic processing and transcription. The development of an accurate biochemical assay for DPC isolation is a priority for the mechanistic understanding of their repair. Here, we propose the STAR assay for the direct quantification of DPCs, sensitive to physiologically relevant treatment conditions. Implementing the STAR assay revealed the formation of small cross-linked peptides on DNA, created by the proteolytic degradation of DPCs by SPRTN. The initial proteolytic degradation of DPCs is required for the downstream activation of DNA repair, which is mediated through the phosphorylation of H2Ax. This leads to the accumulation of DNA repair factors on chromatin and the subsequent complete removal of the cross-linked peptides. These results confirmed that the repair of DPCs is a two-step process, starting with proteolytic resection by SPRTN, followed by the repair of the underlying damage to the DNA.

引用

页数：18

共 64 条

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