The direct interaction of NME3 with Tip60 in DNA repair

2016 ◽  
Vol 473 (9) ◽  
pp. 1237-1245 ◽  
Author(s):  
Ning Tsao ◽  
Ya-Chi Yang ◽  
Yu-Jyun Deng ◽  
Zee-Fen Chang
Keyword(s):  
Dna Damage ◽  
Dna Repair ◽  
Direct Interaction ◽  
Ndp Kinase ◽  
Damage Site ◽  
Site Specific ◽  
Catalytic Function ◽  
Repair Efficiency ◽  
Subsequent Step ◽  
Dna Replication And Repair

Cellular supply of dNTPs via RNR (ribonucleotide reductase) is crucial for DNA replication and repair. It has been shown that DNA-damage-site-specific recruitment of RNR is critical for DNA repair efficiency in quiescent cells. The catalytic function of RNR produces dNDPs. The subsequent step of dNTP formation requires the function of NDP kinase. There are ten isoforms of NDP kinase in human cells. In the present study, we identified NME3 as one specific NDP kinase that interacts directly with Tip60, a histone acetyltransferase, to form a complex with RNR. Our data reveal that NME3 recruitment to DNA damage sites depends on this interaction. Disruption of interaction of NME3 with Tip60 suppressed DNA repair in serum-deprived cells. Thus Tip60 interacts with RNR and NME3 to provide site-specific synthesis of dNTP for facilitating DNA repair in serum-deprived cells which contain low levels of dNTPs.

scholarly journals Identification of MCM8IP, an interactor of MCM8-9 and RPA1 that promotes homologous recombination and DNA synthesis in response to DNA damage

2019 ◽  
Author(s):  
Jen-Wei Huang ◽  
Angelo Taglialatela ◽  
Ananya Acharya ◽  
Giuseppe Leuzzi ◽  
Tarun S. Nambiar ◽  
...  
Keyword(s):  
Dna Damage ◽  
Dna Repair ◽  
Homologous Recombination ◽  
Dna Synthesis ◽  
Dna Recombination ◽  
Direct Interaction ◽  
Response To Treatment ◽  
Parp Inhibition ◽  
Dna Double Strand Breaks ◽  
Crosslinking Agents

ABSTRACTHomologous recombination (HR) mediates the error-free repair of DNA double-strand breaks to maintain genomic stability. HR is carried out by a complex network of DNA repair factors. Here we identify C17orf53/MCM8IP, an OB-fold containing protein that binds ssDNA, as a novel DNA repair factor involved in HR. MCM8IP-deficient cells exhibit HR defects, especially in long-tract gene conversion, occurring downstream of RAD51 loading, consistent with a role for MCM8IP in HR-dependent DNA synthesis. Moreover, loss of MCM8IP confers cellular sensitivity to crosslinking agents and PARP inhibition. Importantly, we identify a direct interaction with MCM8-9, a putative helicase complex mutated in Primary Ovarian Insufficiency, that is crucial for MCM8IP’s ability to promote resistance to DNA damaging agents. In addition to its association with MCM8-9, MCM8IP also binds directly to RPA1. We show that the interactions of MCM8IP with both MCM8-9 and RPA are required to maintain replication fork progression in response to treatment with crosslinking agents. Collectively, our work identifies MCM8IP as a key regulator of DNA damage-associated DNA synthesis during DNA recombination and replication.

DNA repair and genome integrity

2019 ◽  
pp. 13-32
Author(s):  
Giacomo Buscemi
Keyword(s):  
Dna Damage ◽  
Dna Repair ◽  
Genome Stability ◽  
Chemotherapeutic Agents ◽  
Cell Life ◽  
Repair Efficiency ◽  
Damage Response ◽  
Promote Cell Survival ◽  
A Cell ◽  
Repair Ability

The DNA damage response (DDR) is a complex network of pathways involving hundreds of proteins with the main goal to detect and fix lesions occurring to DNA structure, thus preserving genome stability throughout generations. To enhance repair efficiency and eventually clear unrepaired harmful cells, the DDR has under its own control the progression of cell cycle, the induction of cellular senescence and the apoptotic programme. Furthermore, cells take advantage of DDR to manage break-like structures, such as telomeres, and to check processes involving DNA ‘cut and paste’ steps like meiosis and immune response. Since all these aspects of a cell life are frequently altered in cancer, not unexpectedly, deregulation of DDR is an essential step during carcinogenesis. Indeed, even if mutations in DDR genes partially reduce the repair ability of a precancerous cell, they also enhance the possibility of oncogene mutation, allow hyper-replication and promote cell survival and adaptation in stressed conditions. On the other side, impairment of DNA repair sensitizes cancer cells to radio and chemotherapeutic agents inducing DNA damage and DDR components are promising targets to enhance therapy efficiency.

scholarly journals SPRTN protease-cleaved MRE11 decreases DNA repair and radiosensitises cancer cells

2021 ◽  
Vol 12 (2) ◽  
Author(s):  
Juri Na ◽  
Joseph A. Newman ◽  
Chee Kin Then ◽  
Junetha Syed ◽  
Iolanda Vendrell ◽  
...  
Keyword(s):  
Dna Damage ◽  
Dna Repair ◽  
Site Directed Mutagenesis ◽  
Core Complex ◽  
Truncated Form ◽  
Mrn Complex ◽  
The Core ◽  
Repair Efficiency ◽  
Damage Response

AbstractThe human MRE11/RAD50/NBS1 (MRN) complex plays a crucial role in sensing and repairing DNA DSB. MRE11 possesses dual 3′−5′ exonuclease and endonuclease activity and forms the core of the multifunctional MRN complex. We previously identified a C-terminally truncated form of MRE11 (TR-MRE11) associated with post-translational MRE11 degradation. Here we identified SPRTN as the essential protease for the formation of TR-MRE11 and characterised the role of this MRE11 form in its DNA damage response (DDR). Using tandem mass spectrometry and site-directed mutagenesis, the SPRTN-dependent cleavage site for MRE11 was identified between 559 and 580 amino acids. Despite the intact interaction of TR-MRE11 with its constitutive core complex proteins RAD50 and NBS1, both nuclease activities of truncated MRE11 were dramatically reduced due to its deficient binding to DNA. Furthermore, lack of the MRE11 C-terminal decreased HR repair efficiency, very likely due to abolished recruitment of TR-MRE11 to the sites of DNA damage, which consequently led to increased cellular radiosensitivity. The presence of this DNA repair-defective TR-MRE11 could explain our previous finding that the high MRE11 protein expression by immunohistochemistry correlates with improved survival following radical radiotherapy in bladder cancer patients.

Construction of a damage site-specific fluorescent biosensor for single-molecule detection of DNA damage

Talanta ◽  
2021 ◽  
pp. 122809
Author(s):  
Yan Zhang ◽  
Yun Han ◽  
Xiaoran Zou ◽  
Qinfeng Xu ◽  
Fei Ma ◽  
...  
Keyword(s):  
Dna Damage ◽  
Single Molecule ◽  
Single Molecule Detection ◽  
Damage Site ◽  
Site Specific ◽  
Fluorescent Biosensor

scholarly journals SENSITIVITY TO 4-HYDROXYESTRADIOL AND DNA REPAIR EFFICIENCY IN PERIPHERAL BLOOD LYMPHOCYTES OF ENDOMETRIAL CANCER PATIENTS

2018 ◽  
Vol 40 (1) ◽  
pp. 68-72 ◽  
Author(s):  
L G Buchynska ◽  
O V Brieieva
Keyword(s):  
Dna Damage ◽  
Dna Repair ◽  
Endometrial Cancer ◽  
Family History ◽  
Peripheral Blood ◽  
Peripheral Blood Lymphocytes ◽  
Blood Lymphocytes ◽  
Repair Efficiency ◽  
History Of ◽  
History Of Cancer

Background: The development of hormone-dependent cancers, including endometrial carcinomas, in great part may be mediated by the genotoxic effects of estrogen metabolites, among which 4-hydroxyestradiol (4OHE2) is characterized by the most prominent DNA-damaging properties. It is assumed that the individual sensitivity to the 4OHE2 may determine the predisposition to endometrial cancer (EС). Aim: To analyze the sensitivity of peripheral blood lymphocytes (PBLs) of EC patients to the 4OHE2 and to evaluate the repair efficiency of 4OHE2-induced DNA damage. Materials and Methods: The study was performed on the PBLs of 53 EC patients and 20 healthy women. The level of DNA damage was measured using the comet assay and was expressed as % tail DNA. The DNA repair efficiency (%) was evaluated by determining the ratio between the amount of repaired DNA damage and the level of 4OHE2-induced damage that appeared after incubation of PBLs with 4OHE2. Results: In PBLs of EC patients, a higher level of 4OHE2-induced DNA damage (32.0 ± 2.2% tail DNA) and lower DNA repair efficiency (34.0 ± 4.5%) was observed compared to PBLs of healthy women (22.3 ± 2.3% tail DNA and 48.8 ± 4.5%, respectively). PBLs of EC patients with deep tumor invasion of myometrium were characterized by more prominent decrease of DNA repair than those with less invasive tumor (< ½ of myometrium) (20.9 ± 7.8 and 43.7 ± 6.7%, respectively). Furthermore, lower DNA repair efficiency was detected in the PBLs of EC patients with a family history of cancer compared to this parameter in patients with sporadic tumors (20.9±7.8 and 47.1 ± 5.5%, respectively). Conclusion: The PBLs of EC patients are characterized by increased sensitivity to the genotoxic effect of 4OHE2 and reduced repair efficiency regarding 4OHE2-induced DNA damage. A lower level of DNA repair is observed in EC patients with deep tumor myometrial invasion and a family history of cancer.

scholarly journals Activation of PARP2/ARTD2 by DNA damage induces conformational changes relieving enzyme autoinhibition

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Ezeogo Obaji ◽  
Mirko M. Maksimainen ◽  
Albert Galera-Prat ◽  
Lari Lehtiö
Keyword(s):  
Crystal Structure ◽  
Dna Damage ◽  
Dna Repair ◽  
Conformational Changes ◽  
Regulatory Domain ◽  
Damage Site ◽  
Adp Ribosylation ◽  
Adp Ribosyltransferase ◽  
Dna Ends ◽  
Binding Substrate

AbstractHuman PARP2/ARTD2 is an ADP-ribosyltransferase which, when activated by 5′-phosphorylated DNA ends, catalyses poly-ADP-ribosylation of itself, other proteins and DNA. In this study, a crystal structure of PARP2 in complex with an activating 5′-phosphorylated DNA shows that the WGR domain bridges the dsDNA gap and joins the DNA ends. This DNA binding results in major conformational changes, including reorganization of helical fragments, in the PARP2 regulatory domain. A comparison of PARP1 and PARP2 crystal structures reveals how binding to a DNA damage site leads to formation of a catalytically competent conformation. In this conformation, PARP2 is capable of binding substrate NAD+ and histone PARylation factor 1 that changes PARP2 residue specificity from glutamate to serine when initiating DNA repair processes. The structure also reveals how the conformational changes in the autoinhibitory regulatory domain would promote the flexibility needed by the enzyme to reach the target macromolecule for ADP-ribosylation.

scholarly journals Activation of ARTD2/PARP2 by DNA damage induces conformational changes relieving enzyme autoinhibition

2020 ◽  
Author(s):  
Ezeogo Obaji ◽  
Mirko M. Maksimainen ◽  
Albert Galera-Prat ◽  
Lari Lehtiö
Keyword(s):  
Crystal Structure ◽  
Dna Damage ◽  
Dna Repair ◽  
Conformational Changes ◽  
Regulatory Domain ◽  
Damage Site ◽  
Adp Ribosylation ◽  
Adp Ribosyltransferase ◽  
Dna Ends ◽  
Binding Substrate

AbstractHuman ARTD2/PARP2 is an ADP-ribosyltransferase which, when activated by 5’- phosphorylated DNA ends, catalyzes poly-ADP-ribosylation of itself, other proteins and DNA. A crystal structure of ARTD2 in complex with an activating 5’-phosphorylated DNA shows that the WGR domain bridges the dsDNA gap and joins the DNA ends. This DNA binding results in major conformational changes, reorganization of helical fragments, in the ARTD2 regulatory domain. Comparison of ARTD1-3 crystal structures reveal how binding to a DNA damage site leads to formation of a catalytically competent conformation capable of binding substrate NAD+ and histone PARylation factor 1 changing the ARTD2 residue specificity from glutamate to serine when initiating DNA repair processes. The structure also reveals how the conformational changes in the autoinhibitory regulatory domain would promote the flexibility needed by the enzyme to reach the target macromolecule for ADP-ribosylation.

scholarly journals DICER- and MMSET-catalyzed H4K20me2 recruits the nucleotide excision repair factor XPA to DNA damage sites

2017 ◽  
Vol 217 (2) ◽  
pp. 527-540 ◽  
Author(s):  
Shalaka Chitale ◽  
Holger Richly
Keyword(s):  
Dna Damage ◽  
Dna Repair ◽  
Nucleotide Excision Repair ◽  
Excision Repair ◽  
Uv Light ◽  
Dna Lesions ◽  
Histone H4 ◽  
Damage Site ◽  
Nucleotide Excision ◽  
Lesion Recognition

Ultraviolet (UV) irradiation triggers the recruitment of DNA repair factors to the lesion sites and the deposition of histone marks as part of the DNA damage response. The major DNA repair pathway removing DNA lesions caused by exposure to UV light is nucleotide excision repair (NER). We have previously demonstrated that the endoribonuclease DICER facilitates chromatin decondensation during lesion recognition in the global-genomic branch of NER. Here, we report that DICER mediates the recruitment of the methyltransferase MMSET to the DNA damage site. We show that MMSET is required for efficient NER and that it catalyzes the dimethylation of histone H4 at lysine 20 (H4K20me2). H4K20me2 at DNA damage sites facilitates the recruitment of the NER factor XPA. Our work thus provides evidence for an H4K20me2-dependent mechanism of XPA recruitment during lesion recognition in the global-genomic branch of NER.

scholarly journals XPA: DNA Repair Protein of Significant Clinical Importance

2020 ◽  
Vol 21 (6) ◽  
pp. 2182 ◽  
Author(s):  
Lucia Borszéková Pulzová ◽  
Thomas A. Ward ◽  
Miroslav Chovanec
Keyword(s):  
Dna Damage ◽  
Dna Repair ◽  
Protein Interactions ◽  
Excision Repair ◽  
Protein A ◽  
Clinical Importance ◽  
Chemotherapeutic Agents ◽  
Dna Lesions ◽  
Response To Treatment ◽  
Damage Site

The nucleotide excision repair (NER) pathway is activated in response to a broad spectrum of DNA lesions, including bulky lesions induced by platinum-based chemotherapeutic agents. Expression levels of NER factors and resistance to chemotherapy has been examined with some suggestion that NER plays a role in tumour resistance; however, there is a great degree of variability in these studies. Nevertheless, recent clinical studies have suggested Xeroderma Pigmentosum group A (XPA) protein, a key regulator of the NER pathway that is essential for the repair of DNA damage induced by platinum-based chemotherapeutics, as a potential prognostic and predictive biomarker for response to treatment. XPA functions in damage verification step in NER, as well as a molecular scaffold to assemble other NER core factors around the DNA damage site, mediated by protein–protein interactions. In this review, we focus on the interacting partners and mechanisms of regulation of the XPA protein. We summarize clinical oncology data related to this DNA repair factor, particularly its relationship with treatment outcome, and examine the potential of XPA as a target for small molecule inhibitors.

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