In vitro eradication of abasic site-mediated DNA–peptide/protein cross-links by Escherichia coli long-patch base excision repair

Bryan, Cameron; Wei, Xiaoying; Wang, Zhishuo; Yang, Kun

doi:10.1016/j.jbc.2022.102055

Cited by 3 publications

(1 citation statement)

References 62 publications

(103 reference statements)

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“…It should be noted that in addition to DPC MdG , a minor amount (<25%) of DPCs detected from MMS treatment of Chinese hamster lung cells (V79) were attributed to the reaction of AP sites that are formed via depurination of alkylated purines (Scheme ). Recently, additional DPC formation pathways emanating from AP sites have been discovered. − Although SPRTN is involved in the repair of DPCs between AP and the HMCES protein, it is not known whether cross-links between AP sites and other proteins are substrates for the protease as well. , Hence, we cannot rule out cytotoxic contributions by AP-derived DPCs upon MMS treatment. We propose that cells containing defects in DPC repair are profoundly impacted by the formation of approximately 1000 DPC MdG molecules per cell (∼9 DPC MdG /10 8 nt).…”

Section: Discussionmentioning

confidence: 99%

Quantification of Intracellular DNA–Protein Cross-Links with N7-Methyl-2′-Deoxyguanosine and Their Contribution to Cytotoxicity

Wen,

Zhao,

Stingele

et al. 2024

Chem. Res. Toxicol.

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The major product of DNA-methylating agents, N7methyl-2′-deoxyguanosine (MdG), is a persistent lesion in vivo, but it is not believed to have a large direct physiological impact. However, MdG reacts with histone proteins to form reversible DNA−protein cross-links (DPC MdG ), a family of DNA lesions that can significantly threaten cell survival. In this paper, we developed a tandem mass spectrometry method for quantifying the amounts of MdG and DPC MdG in nuclear DNA by taking advantage of their chemical lability and the concurrent release of N7-methylguanine. Using this method, we determined that DPC MdG is formed in less than 1% yield based upon the levels of MdG in methyl methanesulfonate (MMS)-treated HeLa cells. Despite its low chemical yield, DPC MdG contributes to MMS cytotoxicity. Consequently, cells that lack efficient DPC repair by the DPC protease SPRTN are hypersensitive to MMS. This investigation shows that the downstream chemical and biochemical effects of initially formed DNA damage can have significant biological consequences. With respect to MdG formation, the initial DNA lesion is only the beginning.

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Section: Discussionmentioning

confidence: 99%

Quantification of Intracellular DNA–Protein Cross-Links with N7-Methyl-2′-Deoxyguanosine and Their Contribution to Cytotoxicity

Wen,

Zhao,

Stingele

et al. 2024

Chem. Res. Toxicol.

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Saccharomyces cerevisiae apurinic/apyrimidinic endonuclease 1 repairs abasic site-mediated DNA-peptide/protein cross-links

Bryan

Wei

et al. 2023

DNA Repair

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Novel mechanisms for the removal of strong replication-blocking HMCES- and thiazolidine-DNA adducts in humans

Sugimoto

Masuda

Iwai

et al. 2023

Nucleic Acids Research

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Apurinic/apyrimidinic (AP) sites are DNA lesions created under normal growth conditions that result in cytotoxicity, replication-blocks, and mutations. AP sites are susceptible to β-elimination and are liable to be converted to DNA strand breaks. HMCES (5-hydroxymethylcytosine binding, ES cell specific) protein interacts with AP sites in single stranded (ss) DNA exposed at DNA replication forks to generate a stable thiazolidine protein-DNA crosslink and protect cells against AP site toxicity. The crosslinked HMCES is resolved by proteasome-mediated degradation; however, it is unclear how HMCES-crosslinked ssDNA and the resulting proteasome-degraded HMCES adducts are processed and repaired. Here, we describe methods for the preparation of thiazolidine adduct-containing oligonucleotides and determination of their structure. We demonstrate that the HMCES-crosslink is a strong replication blocking adduct and that protease-digested HMCES adducts block DNA replication to a similar extent as AP sites. Moreover, we show that the human AP endonuclease APE1 incises DNA 5′ to the protease-digested HMCES adduct. Interestingly, while HMCES-ssDNA crosslinks are stable, the crosslink is reversed upon the formation of dsDNA, possibly due to a catalytic reverse reaction. Our results shed new light on damage tolerance and repair pathways for HMCES-DNA crosslinks in human cells.

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In vitro eradication of abasic site-mediated DNA–peptide/protein cross-links by Escherichia coli long-patch base excision repair

Cited by 3 publications

References 62 publications

Quantification of Intracellular DNA–Protein Cross-Links with N7-Methyl-2′-Deoxyguanosine and Their Contribution to Cytotoxicity

Quantification of Intracellular DNA–Protein Cross-Links with N7-Methyl-2′-Deoxyguanosine and Their Contribution to Cytotoxicity

Saccharomyces cerevisiae apurinic/apyrimidinic endonuclease 1 repairs abasic site-mediated DNA-peptide/protein cross-links

Novel mechanisms for the removal of strong replication-blocking HMCES- and thiazolidine-DNA adducts in humans

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