Mitotic DNA synthesis is caused by transcription-replication conflicts in BRCA2-deficient cells
Aberrant replication causes cells lacking BRCA2 to enter mitosis with under-replicated DNA, which activates a repair mechanism known as mitotic DNA synthesis (MiDAS). Here, we identify genome-wide the sites where MiDAS reactions occur when BRCA2 is abrogated. High-resolution profiling revealed that...
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| Published in: | Molecular cell Vol. 82; no. 18; p. 3382 |
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| Main Authors: | , , , , , , , , , , , , , , , , , , |
| Format: | Journal Article |
| Language: | English |
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15.09.2022
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| ISSN: | 1097-4164, 1097-4164 |
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| Abstract | Aberrant replication causes cells lacking BRCA2 to enter mitosis with under-replicated DNA, which activates a repair mechanism known as mitotic DNA synthesis (MiDAS). Here, we identify genome-wide the sites where MiDAS reactions occur when BRCA2 is abrogated. High-resolution profiling revealed that these sites are different from MiDAS at aphidicolin-induced common fragile sites in that they map to genomic regions replicating in the early S-phase, which are close to early-firing replication origins, are highly transcribed, and display R-loop-forming potential. Both transcription inhibition in early S-phase and RNaseH1 overexpression reduced MiDAS in BRCA2-deficient cells, indicating that transcription-replication conflicts (TRCs) and R-loops are the source of MiDAS. Importantly, the MiDAS sites identified in BRCA2-deficient cells also represent hotspots for genomic rearrangements in BRCA2-mutated breast tumors. Thus, our work provides a mechanism for how tumor-predisposing BRCA2 inactivation links transcription-induced DNA damage with mitotic DNA repair to fuel the genomic instability characteristic of cancer cells. |
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| AbstractList | Aberrant replication causes cells lacking BRCA2 to enter mitosis with under-replicated DNA, which activates a repair mechanism known as mitotic DNA synthesis (MiDAS). Here, we identify genome-wide the sites where MiDAS reactions occur when BRCA2 is abrogated. High-resolution profiling revealed that these sites are different from MiDAS at aphidicolin-induced common fragile sites in that they map to genomic regions replicating in the early S-phase, which are close to early-firing replication origins, are highly transcribed, and display R-loop-forming potential. Both transcription inhibition in early S-phase and RNaseH1 overexpression reduced MiDAS in BRCA2-deficient cells, indicating that transcription-replication conflicts (TRCs) and R-loops are the source of MiDAS. Importantly, the MiDAS sites identified in BRCA2-deficient cells also represent hotspots for genomic rearrangements in BRCA2-mutated breast tumors. Thus, our work provides a mechanism for how tumor-predisposing BRCA2 inactivation links transcription-induced DNA damage with mitotic DNA repair to fuel the genomic instability characteristic of cancer cells.Aberrant replication causes cells lacking BRCA2 to enter mitosis with under-replicated DNA, which activates a repair mechanism known as mitotic DNA synthesis (MiDAS). Here, we identify genome-wide the sites where MiDAS reactions occur when BRCA2 is abrogated. High-resolution profiling revealed that these sites are different from MiDAS at aphidicolin-induced common fragile sites in that they map to genomic regions replicating in the early S-phase, which are close to early-firing replication origins, are highly transcribed, and display R-loop-forming potential. Both transcription inhibition in early S-phase and RNaseH1 overexpression reduced MiDAS in BRCA2-deficient cells, indicating that transcription-replication conflicts (TRCs) and R-loops are the source of MiDAS. Importantly, the MiDAS sites identified in BRCA2-deficient cells also represent hotspots for genomic rearrangements in BRCA2-mutated breast tumors. Thus, our work provides a mechanism for how tumor-predisposing BRCA2 inactivation links transcription-induced DNA damage with mitotic DNA repair to fuel the genomic instability characteristic of cancer cells. Aberrant replication causes cells lacking BRCA2 to enter mitosis with under-replicated DNA, which activates a repair mechanism known as mitotic DNA synthesis (MiDAS). Here, we identify genome-wide the sites where MiDAS reactions occur when BRCA2 is abrogated. High-resolution profiling revealed that these sites are different from MiDAS at aphidicolin-induced common fragile sites in that they map to genomic regions replicating in the early S-phase, which are close to early-firing replication origins, are highly transcribed, and display R-loop-forming potential. Both transcription inhibition in early S-phase and RNaseH1 overexpression reduced MiDAS in BRCA2-deficient cells, indicating that transcription-replication conflicts (TRCs) and R-loops are the source of MiDAS. Importantly, the MiDAS sites identified in BRCA2-deficient cells also represent hotspots for genomic rearrangements in BRCA2-mutated breast tumors. Thus, our work provides a mechanism for how tumor-predisposing BRCA2 inactivation links transcription-induced DNA damage with mitotic DNA repair to fuel the genomic instability characteristic of cancer cells. |
| Author | Petropoulos, Michalis Panagopoulos, Andreas Paulsen, Teressa Dagg, Rebecca A Ochs, Fena Karamichali, Angeliki Puig Lombardi, Emilia Jones, Samuel E Halazonetis, Thanos D Groelly, Florian J Dionellis, Vasilis S Prasad, Birbal Tarsounas, Madalena Miossec, Matthieu J Rossetti, Giacomo G Lockstone, Helen Blackford, Andrew N Legube, Gaëlle Altmeyer, Matthias |
| Author_xml | – sequence: 1 givenname: Florian J surname: Groelly fullname: Groelly, Florian J organization: Genome Stability and Tumourigenesis Group, Department of Oncology, Oxford Institute for Radiation Oncology, University of Oxford, Oxford OX3 7DQ, UK – sequence: 2 givenname: Rebecca A surname: Dagg fullname: Dagg, Rebecca A organization: Genome Stability and Tumourigenesis Group, Department of Oncology, Oxford Institute for Radiation Oncology, University of Oxford, Oxford OX3 7DQ, UK – sequence: 3 givenname: Michalis surname: Petropoulos fullname: Petropoulos, Michalis organization: Department of Molecular Biology, University of Geneva, 1205 Geneva, Switzerland – sequence: 4 givenname: Giacomo G surname: Rossetti fullname: Rossetti, Giacomo G organization: Department of Molecular Biology, University of Geneva, 1205 Geneva, Switzerland – sequence: 5 givenname: Birbal surname: Prasad fullname: Prasad, Birbal organization: Genome Stability and Tumourigenesis Group, Department of Oncology, Oxford Institute for Radiation Oncology, University of Oxford, Oxford OX3 7DQ, UK – sequence: 6 givenname: Andreas surname: Panagopoulos fullname: Panagopoulos, Andreas organization: Department of Molecular Mechanisms of Disease, University of Zurich, 8057 Zurich, Switzerland – sequence: 7 givenname: Teressa surname: Paulsen fullname: Paulsen, Teressa organization: Genome Stability and Tumourigenesis Group, Department of Oncology, Oxford Institute for Radiation Oncology, University of Oxford, Oxford OX3 7DQ, UK – sequence: 8 givenname: Angeliki surname: Karamichali fullname: Karamichali, Angeliki organization: Department of Molecular Biology, University of Geneva, 1205 Geneva, Switzerland – sequence: 9 givenname: Samuel E surname: Jones fullname: Jones, Samuel E organization: Department of Oncology, MRC Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DS, UK – sequence: 10 givenname: Fena surname: Ochs fullname: Ochs, Fena organization: Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK – sequence: 11 givenname: Vasilis S surname: Dionellis fullname: Dionellis, Vasilis S organization: Department of Molecular Biology, University of Geneva, 1205 Geneva, Switzerland – sequence: 12 givenname: Emilia surname: Puig Lombardi fullname: Puig Lombardi, Emilia organization: Genome Stability and Tumourigenesis Group, Department of Oncology, Oxford Institute for Radiation Oncology, University of Oxford, Oxford OX3 7DQ, UK – sequence: 13 givenname: Matthieu J surname: Miossec fullname: Miossec, Matthieu J organization: Bioinformatics and Statistical Genetics Core, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford OX3 7BN, UK – sequence: 14 givenname: Helen surname: Lockstone fullname: Lockstone, Helen organization: Bioinformatics and Statistical Genetics Core, Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford OX3 7BN, UK – sequence: 15 givenname: Gaëlle surname: Legube fullname: Legube, Gaëlle organization: LBCMCP, Centre de Biologie Intégrative (CBI), CNRS, Université de Toulouse, UT3, Toulouse 31062, France – sequence: 16 givenname: Andrew N surname: Blackford fullname: Blackford, Andrew N organization: Department of Oncology, MRC Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DS, UK – sequence: 17 givenname: Matthias surname: Altmeyer fullname: Altmeyer, Matthias organization: Department of Molecular Mechanisms of Disease, University of Zurich, 8057 Zurich, Switzerland – sequence: 18 givenname: Thanos D surname: Halazonetis fullname: Halazonetis, Thanos D email: thanos.halazonetis@unige.ch organization: Department of Molecular Biology, University of Geneva, 1205 Geneva, Switzerland. Electronic address: thanos.halazonetis@unige.ch – sequence: 19 givenname: Madalena surname: Tarsounas fullname: Tarsounas, Madalena email: madalena.tarsounas@oncology.ox.ac.uk organization: Genome Stability and Tumourigenesis Group, Department of Oncology, Oxford Institute for Radiation Oncology, University of Oxford, Oxford OX3 7DQ, UK. Electronic address: madalena.tarsounas@oncology.ox.ac.uk |
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| Keywords | genome stability transcription-replication conflicts MiDAS TRCs R-loops mitotic DNA synthesis BRCA2 |
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| SubjectTerms | Aphidicolin - pharmacology BRCA2 Protein - genetics Chromosome Fragile Sites - genetics DNA - genetics DNA Damage DNA Replication Genomic Instability Humans Mitosis - genetics |
| Title | Mitotic DNA synthesis is caused by transcription-replication conflicts in BRCA2-deficient cells |
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