المؤلفون: Shunichi Takeda, Markus Löbrich, Olivia Barton, Andrea Beucher, Verena Quennet

المصدر: Nucleic Acids Research

مصطلحات موضوعية: Threonine, DNA Repair, DNA repair, Cell Cycle Proteins, Genome Integrity, Repair and Replication, Biology, chemistry.chemical_compound, MRE11 Homologue Protein, Genetics, Humans, DNA Breaks, Double-Stranded, Phosphorylation, Cells, Cultured, Etoposide, Endodeoxyribonucleases, Topoisomerase, fungi, G1 Phase, Nuclear Proteins, Antineoplastic Agents, Phytogenic, Molecular biology, DNA-Binding Proteins, Non-homologous end joining, enzymes and coenzymes (carbohydrates), MRX complex, chemistry, MRN complex, biology.protein, Carrier Proteins, Homologous recombination, DNA

الوصف: Topoisomerases class II (topoII) cleave and re-ligate the DNA double helix to allow the passage of an intact DNA strand through it. Chemotherapeutic drugs such as etoposide target topoII, interfere with the normal enzymatic cleavage/re-ligation reaction and create a DNA double-strand break (DSB) with the enzyme covalently bound to the 5���-end of the DNA. Such DSBs are repaired by one of the two major DSB repair pathways, non-homologous end-joining (NHEJ) or homologous recombination. However, prior to repair, the covalently bound topoII needs to be removed from the DNA end, a process requiring the MRX complex and ctp1 in fission yeast. CtIP, the mammalian ortholog of ctp1, is known to promote homologous recombination by resecting DSB ends. Here, we show that human cells arrested in G0/G1 repair etoposide-induced DSBs by NHEJ and, surprisingly, require the MRN complex (the ortholog of MRX) and CtIP. CtIP's function for repairing etoposide-induced DSBs by NHEJ in G0/G1 requires the Thr-847 but not the Ser-327 phosphorylation site, both of which are needed for resection during HR. This finding establishes that CtIP promotes NHEJ of etoposide-induced DSBs during G0/G1 phase with an end-processing function that is distinct to its resection function.

وصف الملف: text

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::b6ddb016ab3d9de2a39c9569151e4329Test

المؤلفون: Jordane Dépagne, Xavier Veaute, Florian Roisné-Hamelin, Sophie Zinn-Justin, Stéphane Marcand, Sabrina Pobiega, Didier Busso, Marie-Hélène Le Du, Philippe Cuniasse, Jean-Baptiste Charbonnier, Simona Miron, Kévin Jézéquel, Isabelle Callebaut

المساهمون: Institut de Biologie François JACOB (JACOB), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Institut de Radiobiologie Cellulaire et Moléculaire (IRCM), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Stabilité génétique, cellules souches et radiations (SGCSR (U_1274 / UMR_E_008)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Saclay-Université Paris Cité (UPCité), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Institut de minéralogie, de physique des matériaux et de cosmochimie (IMPMC), Muséum national d'Histoire naturelle (MNHN)-Institut de recherche pour le développement [IRD] : UR206-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Saclay-Université de Paris (UP), Gestionnaire, Hal Sorbonne Université

المصدر: Nature Communications
Nature Communications, 2021, 12 (1), pp.2763. ⟨10.1038/s41467-021-23035-w⟩
Nature Communications, Nature Publishing Group, 2021, 12 (1), pp.2763. ⟨10.1038/s41467-021-23035-w⟩
Nature Communications, Vol 12, Iss 1, Pp 1-16 (2021)

مصطلحات موضوعية: 0301 basic medicine, Models, Molecular, DNA End-Joining Repair, Saccharomyces cerevisiae Proteins, Science, [SDV]Life Sciences [q-bio], Saccharomyces cerevisiae, Amino Acid Motifs, Telomere-Binding Proteins, General Physics and Astronomy, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Protein Domains, DNA Breaks, Double-Stranded, DNA, Fungal, Multidisciplinary, Endodeoxyribonucleases, biology, Transition (genetics), Chemistry, Chromosome, General Chemistry, Telomere, biology.organism_classification, Cell biology, DNA-Binding Proteins, [SDV] Life Sciences [q-bio], 030104 developmental biology, MRX complex, Exodeoxyribonucleases, Docking (molecular), Rad50, Multiprotein Complexes, Mutation, Chromosomes, Fungal, 030217 neurology & neurosurgery, DNA, Protein Binding

الوصف: Specific proteins present at telomeres ensure chromosome end stability, in large part through unknown mechanisms. In this work, we address how the Saccharomyces cerevisiae ORC-related Rif2 protein protects telomere. We show that the small N-terminal Rif2 BAT motif (Blocks Addition of Telomeres) previously known to limit telomere elongation and Tel1 activity is also sufficient to block NHEJ and 5’ end resection. The BAT motif inhibits the ability of the Mre11-Rad50-Xrs2 complex (MRX) to capture DNA ends. It acts through a direct contact with Rad50 ATP-binding Head domains. Through genetic approaches guided by structural predictions, we identify residues at the surface of Rad50 that are essential for the interaction with Rif2 and its inhibition. Finally, a docking model predicts how BAT binding could specifically destabilise the DNA-bound state of the MRX complex. From these results, we propose that when an MRX complex approaches a telomere, the Rif2 BAT motif binds MRX Head in its ATP-bound resting state. This antagonises MRX transition to its DNA-bound state, and favours a rapid return to the ATP-bound state. Unable to stably capture the telomere end, the MRX complex cannot proceed with the subsequent steps of NHEJ, Tel1-activation and 5’ resection.

وصف الملف: application/pdf

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::dea365aff9a28535649b01f33e0d9ab1Test
https://hal.sorbonne-universite.fr/hal-03231067/documentTest

المؤلفون: Maria Pia Longhese, Jacopo Vertemara, Renata Tisi, Giuseppe Zampella

المساهمون: Tisi, R, Vertemara, J, Zampella, G, Longhese, M

المصدر: Computational and Structural Biotechnology Journal
Computational and Structural Biotechnology Journal, Vol 18, Iss, Pp 1137-1152 (2020)

مصطلحات موضوعية: CHIM/03 - CHIMICA GENERALE ED INORGANICA, DNA damage, lcsh:Biotechnology, Biophysics, BIO/18 - GENETICA, MRX/MRN, Review Article, Biology, Biochemistry, DNA sequencing, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Structural Biology, lcsh:TP248.13-248.65, Mre11, Genetics, Homologous chromosome, ComputingMethodologies_COMPUTERGRAPHICS, 030304 developmental biology, 0303 health sciences, Xrs2/NBS1, BIO/11 - BIOLOGIA MOLECOLARE, Double-strand break (DSB), Computer Science Applications, Cell biology, enzymes and coenzymes (carbohydrates), MRX complex, MRN complex, chemistry, 030220 oncology & carcinogenesis, Rad50, Homologous recombination, DNA, Biotechnology

الوصف: Graphical abstract
Chromosomal DNA double-strand breaks (DSBs) are potentially lethal DNA lesions that pose a significant threat to genome stability and therefore need to be repaired to preserve genome integrity. Eukaryotic cells possess two main mechanisms for repairing DSBs: non-homologous end-joining (NHEJ) and homologous recombination (HR). HR requires that the 5′ terminated strands at both DNA ends are nucleolytically degraded by a concerted action of nucleases in a process termed DNA-end resection. This degradation leads to the formation of 3′-ended single-stranded DNA (ssDNA) ends that are essential to use homologous DNA sequences for repair. The evolutionarily conserved Mre11-Rad50-Xrs2/NBS1 complex (MRX/MRN) has enzymatic and structural activities to initiate DSB resection and to maintain the DSB ends tethered to each other for their repair. Furthermore, it is required to recruit and activate the protein kinase Tel1/ATM, which plays a key role in DSB signaling. All these functions depend on ATP-regulated DNA binding and nucleolytic activities of the complex. Several structures have been obtained in recent years for Mre11 and Rad50 subunits from archaea, and a few from the bacterial and eukaryotic orthologs. Nevertheless, the mechanism of activation of this protein complex is yet to be fully elucidated. In this review, we focused on recent biophysical and structural insights on the MRX complex and their interplay.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::21f386ea780002cbd131fdb18d2754f5Test
https://doi.org/10.1016/j.csbj.2020.05.013Test

المؤلفون: Erika Casari, Maria Pia Longhese, Renata Tisi, Corinne Cassani, Antonio Marsella

المساهمون: Marsella, A, Cassani, C, Casari, E, Tisi, R, Longhese, M

المصدر: Current Genetics. 65:11-16

مصطلحات موضوعية: Saccharomyces cerevisiae Proteins, DNA damage, BIO/18 - GENETICA, Saccharomyces cerevisiae, Biology, Proteomics, Resection, Conserved sequence, 03 medical and health sciences, chemistry.chemical_compound, Mre11, Genetics, Allele, DNA, Fungal, Sae2, 030304 developmental biology, 0303 health sciences, Endodeoxyribonucleases, Tel1, 030302 biochemistry & molecular biology, General Medicine, BIO/11 - BIOLOGIA MOLECOLARE, Endonucleases, Cell biology, DNA-Binding Proteins, enzymes and coenzymes (carbohydrates), Exodeoxyribonucleases, MRX complex, chemistry, Rad50, DNA, DNA Damage

الوصف: The evolutionarily conserved Mre11-Rad50-Xrs2 (MRX) complex cooperates with the Sae2 protein in initiating resection of DNA double-strand breaks (DSBs) and in maintaining the DSB ends tethered to each other for their accurate repair. How these MRX-Sae2 functions contribute to DNA damage resistance is not understood. By taking advantage of mre11 alleles that suppress the hypersensitivity of sae2∆ cells to genotoxic agents, we have recently found that Mre11 can be divided in two structurally distinct domains that support resistance to genotoxic agents by mediating different processes. While the Mre11 N-terminal domain impacts on the resection activity of long-range resection nucleases by mediating MRX and Tel1/ATM association to DNA DSBs, the C-terminus influences the MRX-tethering activity by its virtue to interact with Rad50. Given the evolutionary conservation of the MRX complex, our results have implications for understanding the consequences of its dysfunctions in human diseases.

وصف الملف: ELETTRONICO

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::694f5709019ee68008246fa1dccfdcb4Test
https://doi.org/10.1007/s00294-018-0861-5Test

المؤلفون: Marco Gnugnoli, Carlo Rinaldi, Maria Pia Longhese, Antonio Marsella, Erika Casari, Diego Bonetti, Chiara Vittoria Colombo

المساهمون: Casari, E, Rinaldi, C, Marsella, A, Gnugnoli, M, Colombo, C, Bonetti, D, Longhese, M

المصدر: Frontiers in Molecular Biosciences
Frontiers in Molecular Biosciences, Vol 6 (2019)

مصطلحات موضوعية: Programmed cell death, genetic processes, Context (language use), BIO/18 - GENETICA, Review, Biochemistry, Genetics and Molecular Biology (miscellaneous), Biochemistry, Sae2/CtIP, chemistry.chemical_compound, Mre11, Molecular Biosciences, Molecular Biology, lcsh:QH301-705.5, MRX complex, Chemistry, Kinase, fungi, Xrs2/NBS1, Resection, Cell biology, Chromatin, enzymes and coenzymes (carbohydrates), MRN complex, lcsh:Biology (General), Double-strand break, Rad50, Tel1/ATM, biological phenomena, cell phenomena, and immunity, DNA

الوصف: DNA double-strand breaks (DSBs) are highly cytotoxic lesions that must be repaired to ensure genomic stability and avoid cell death. The cellular response to DSBs is initiated by the evolutionarily conserved Mre11-Rad50-Xrs2/NBS1 (MRX/MRN) complex that has structural and catalytic functions. Furthermore, it is responsible for DSB signaling through the activation of the checkpoint kinase Tel1/ATM. Here, we review functions and regulation of the MRX/MRN complex in DSB processing in a chromatin context, as well as its interplay with Tel1/ATM.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::b557c6420b82aef2a287d8dbc8fc06d2Test
http://hdl.handle.net/10281/239891Test

المؤلفون: Adam S. Miller, Yong Xiong, David G. Maranon, Fengshan Liang, Caroline Tang, Patrick Sung, Claudia Wiese, Simonne Longerich, Gary M. Kupfer, Olga Buzovetsky

المصدر: Cell Reports, Vol 15, Iss 10, Pp 2118-2126 (2016)

مصطلحات موضوعية: 0301 basic medicine, DNA Repair, DNA damage, DNA repair, RAD51, Biology, Models, Biological, General Biochemistry, Genetics and Molecular Biology, Article, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Protein Domains, Humans, Amino Acid Sequence, Homologous Recombination, Replication protein A, lcsh:QH301-705.5, Genetics, Nuclear Proteins, DNA, Cell biology, DNA-Binding Proteins, Chromosome Pairing, 030104 developmental biology, MRX complex, MRN complex, chemistry, lcsh:Biology (General), 030220 oncology & carcinogenesis, Rad51 Recombinase, Homologous recombination, DNA Damage, HeLa Cells, Protein Binding

الوصف: SummaryThe UAF1-USP1 complex deubiquitinates FANCD2 during execution of the Fanconi anemia DNA damage response pathway. As such, UAF1 depletion results in persistent FANCD2 ubiquitination and DNA damage hypersensitivity. UAF1-deficient cells are also impaired for DNA repair by homologous recombination. Herein, we show that UAF1 binds DNA and forms a dimeric complex with RAD51AP1, an accessory factor of the RAD51 recombinase, and a trimeric complex with RAD51 through RAD51AP1. Two small ubiquitin-like modifier (SUMO)-like domains in UAF1 and a SUMO-interacting motif in RAD51AP1 mediate complex formation. Importantly, UAF1 enhances RAD51-mediated homologous DNA pairing in a manner that is dependent on complex formation with RAD51AP1 but independent of USP1. Mechanistically, RAD51AP1-UAF1 co-operates with RAD51 to assemble the synaptic complex, a critical nucleoprotein intermediate in homologous recombination, and cellular studies reveal the biological significance of the RAD51AP1-UAF1 protein complex. Our findings provide insights into an apparently USP1-independent role of UAF1 in genome maintenance.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::8960b4484d551fc3b0c69a944780bf5aTest

المؤلفون: Maria Pia Longhese, Patrick Sung, Corinne Cassani, Elisa Gobbini, Fabiana Mambretti, Weibin Wang, Erika Casari, Giuseppe Zampella, Renata Tisi, Jacopo Vertemara

المساهمون: Gobbini, E, Cassani, C, Vertemara, J, Wang, W, Mambretti, F, Casari, E, Sung, P, Tisi, R, Zampella, G, Longhese, M

المصدر: The EMBO Journal. 37

مصطلحات موضوعية: 0301 basic medicine, Saccharomyces cerevisiae Proteins, genetic processes, Mutant, Saccharomyces cerevisiae, Biology, Exo1, MRX, General Biochemistry, Genetics and Molecular Biology, Resection, 03 medical and health sciences, chemistry.chemical_compound, Protein Domains, DNA Breaks, Double-Stranded, resection, DNA, Fungal, Sae2, Molecular Biology, Nuclease, Endodeoxyribonucleases, General Immunology and Microbiology, General Neuroscience, fungi, Articles, double‐strand break, Cell biology, enzymes and coenzymes (carbohydrates), Exodeoxyribonucleases, 030104 developmental biology, MRX complex, chemistry, Multiprotein Complexes, health occupations, biology.protein, biological phenomena, cell phenomena, and immunity, Homologous recombination, Function (biology), DNA

الوصف: Homologous recombination is triggered by nucleolytic degradation (resection) of DNA double‐strand breaks (DSBs). DSB resection requires the Mre11‐Rad50‐Xrs2 (MRX) complex, which promotes the activity of Exo1 nuclease through a poorly understood mechanism. Here, we describe the Mre11‐R10T mutant variant that accelerates DSB resection compared to wild‐type Mre11 by potentiating Exo1‐mediated processing. This increased Exo1 resection activity leads to a decreased association of the Ku complex to DSBs and an enhanced DSB resection in G1, indicating that Exo1 has a direct function in preventing Ku association with DSBs. Molecular dynamics simulations show that rotation of the Mre11 capping domains is able to induce unwinding of double‐strand DNA (dsDNA). The R10T substitution causes altered orientation of the Mre11 capping domain that leads to persistent melting of the dsDNA end. We propose that MRX creates a specific DNA end structure that promotes Exo1 resection activity by facilitating the persistence of this nuclease on the DSB ends, uncovering a novel MRX function in DSB resection.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::95449b2ec079c017da74de7e68e913e8Test
https://doi.org/10.15252/embj.201798588Test

المؤلفون: Miki Shinohara, Kenichiro Matsuzaki

المصدر: Biochemical and biophysical research communications. 501(4)

مصطلحات موضوعية: 0301 basic medicine, DNA End-Joining Repair, Saccharomyces cerevisiae Proteins, Protein subunit, Biophysics, Biochemistry, 03 medical and health sciences, chemistry.chemical_compound, Protein Domains, Catalytic Domain, Amino Acid Sequence, Phosphorylation, Casein Kinase II, Molecular Biology, chemistry.chemical_classification, DNA ligase, Chemistry, fungi, Cell Biology, Recombinant Proteins, Cell biology, Non-homologous end joining, DNA-Binding Proteins, enzymes and coenzymes (carbohydrates), 030104 developmental biology, MRX complex, embryonic structures, Casein kinase 2, Homologous recombination, DNA, Protein Binding

الوصف: A DNA double strand break (DSB) is one of the most cytotoxic DNA lesions, but it can be repaired by non-homologous end joining (NHEJ) or by homologous recombination. The choice between these two repair pathways depends on the cell cycle stage. Although NHEJ constitutes a simple re-ligation reaction, the regulatory mechanism(s) controlling its activity has not been fully characterized. Lif1 is a regulatory subunit of the NHEJ-specific DNA ligase IV and interacts with Xrs2 of the MRX complex which is a key factor in DSB repair. Specifically, the C-terminal region of Lif1, which contains a CK2-specific phosphorylation motif, interacts with the FHA domain of Xrs2 during canonical- NHEJ (C-NHEJ). Herein, we show that Lif1 and Cka2, a catalytic subunit of yeast CK2, interact and that the C-terminal phosphorylation consensus motif in Lif1 is phosphorylated by recombinant CK2. These observations suggest that phosphorylation of Lif1 by CK2 at a DSB site promotes the Lif1-Xrs2 interaction and facilitates C-NHEJ.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::182da1c30a25e394f59d69f01315b5d9Test
https://pubmed.ncbi.nlm.nih.gov/29778533Test

المؤلفون: Michela Clerici, Maria Pia Longhese, Diego Bonetti, Chiara Vittoria Colombo

المساهمون: Bonetti, D, Colombo, C, Clerici, M, Longhese, M

المصدر: Frontiers in Genetics, Vol 9 (2018)

مصطلحات موضوعية: 0301 basic medicine, lcsh:QH426-470, ved/biology.organism_classification_rank.species, Saccharomyces cerevisiae, Biology, DNA replication, medicine.disease_cause, MRX, 03 medical and health sciences, chemistry.chemical_compound, Nuclease, Genetic, Genetics, medicine, Model organism, Genetics (clinical), ved/biology, Checkpoint, fungi, Chromosome, biology.organism_classification, Resection, Cell biology, lcsh:Genetics, 030104 developmental biology, MRX complex, chemistry, Double-strand break, Molecular Medicine, nucleases, Homologous recombination, Carcinogenesis, DNA

الوصف: DNA double-strand breaks (DSBs) are particularly hazardous lesions as their inappropriate repair can result in chromosome rearrangements, an important driving force of tumorigenesis. DSBs can be repaired by end joining mechanisms or by homologous recombination (HR). HR requires the action of several nucleases that preferentially remove the 5'-terminated strands at both DSB ends in a process called DNA end resection. The same nucleases are also involved in the processing of replication fork structures. Much of our understanding of these pathways has come from studies in the model organism Saccharomyces cerevisiae. Here, we review the current knowledge of the mechanism of resection at DNA DSBs and replication forks.

وصف الملف: ELETTRONICO

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::ea568f03b2c46140b64d5797deaaa69cTest
http://hdl.handle.net/10281/213196Test

المؤلفون: Matteo Villa, Giulia Tedeschi, Diego Bonetti, Maria Pia Longhese, Corinne Cassani, Elisa Gobbini

المساهمون: Bonetti, D, Villa, M, Gobbini, E, Cassani, C, Tedeschi, G, Longhese, M

المصدر: EMBO reports. 16:351-361

مصطلحات موضوعية: double-strand break, Saccharomyces cerevisiae Proteins, genetic processes, Mutant, Saccharomyces cerevisiae, BIO/18 - GENETICA, Cell Cycle Proteins, Biology, Models, Biological, Biochemistry, Resection, chemistry.chemical_compound, Genetic, Rad9, Genetics, resection, Molecular Biology, Sgs1, Endodeoxyribonucleases, RecQ Helicases, Scientific Reports, fungi, Recombinational DNA Repair, Endonucleases, biology.organism_classification, Molecular biology, Cell biology, DNA-Binding Proteins, enzymes and coenzymes (carbohydrates), Exodeoxyribonucleases, MRX complex, chemistry, Multiprotein Complexes, Extensive resection, biological phenomena, cell phenomena, and immunity, Homologous recombination, DNA

الوصف: Homologous recombination requires nucleolytic degradation (resection) of DNA double-strand break (DSB) ends. In Saccharomyces cerevisiae, the MRX complex and Sae2 are involved in the onset of DSB resection, whereas extensive resection requires Exo1 and the concerted action of Dna2 and Sgs1. Here, we show that the checkpoint protein Rad9 limits the action of Sgs1/Dna2 in DSB resection by inhibiting Sgs1 binding/persistence at the DSB ends. When inhibition by Rad9 is abolished by the Sgs1-ss mutant variant or by deletion of RAD9, the requirement for Sae2 and functional MRX in DSB resection is reduced. These results provide new insights into how early and long-range resection is coordinated. Synopsis The checkpoint protein Rad9 inhibits the Sgs1/Dna2 long-range resection machinery and thereby increases the requirement for MRX/Sae2 activities in DSB resection. The Sgs1-ss mutant variant suppresses the sensitivity to DNA damaging agents and the resection defect of sae2 cells. Rad9 limits the action of Sgs1/Dna2 in DSB resection by inhibiting Sgs1 binding/persistence at the DSB ends. The escape of Sgs1 from Rad9 inhibition reduces the requirement for Sae2 and functional MRX in DSB resection. The checkpoint protein Rad9 inhibits the Sgs1/Dna2 long-range resection machinery and thereby increases the requirement for MRX-Sae2 activities in DSB resection.

وصف الملف: STAMPA

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::b9964b581dbcdcb2905ef442b825421bTest
https://doi.org/10.15252/embr.201439764Test