Mitotic DNA synthesis is caused by transcription-replication conflicts in BRCA2-deficient cells

Groelly, Florian J.; Dagg, Rebecca A.; Petropoulos, Michalis; Rossetti, Giacomo G.; Prasad, Birbal; Παναγόπουλος, Ανδρέας; Paulsen, Teressa; Karamichali, Angeliki; Jones, Samuel E.; Ochs, Fena; Dionellis, Vasilis S.; Lombardi, Emilia Puig; Miossec, Matthieu J.; Lockstone, Helen; Legube, Gaëlle; Blackford, Andrew N.; Altmeyer, Matthias; Halazonetis, Thanos D.; Tarsounas, Madalena

doi:10.1016/j.molcel.2022.07.011

Cited by 24 publications

(21 citation statements)

References 79 publications

(150 reference statements)

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“…It will be important in the future to understand how FANCD2 functionally interacts with SLX4, and with the other members of the FA pathway, to promote SSE-mediated resolution of these structures. Finally, the synthetic proliferation defect and the difference in the impact of SETX depletion in cancerous versus the noncancerous cell lines hint towards therapeutic strategies targeting endogenous RS in tumors, especially in specific genetic backgrounds [71][72][73] or contexts characterized by increased TRCs, such as following oncogene activation, metabolic stress or estrogenstimulated transcription [74][75][76] . For example, even if germline mutations in SETX associated with neurodegenerative disorders do not reportedly increase cancer risk, SETX somatic mutations and CNVs are commonly found in tumors, particularly in gynecological and colon cancers (data from The Cancer Genome Atlas (TGCA) Program) 77 .…”

Section: Discussionmentioning

confidence: 99%

FANCD2 promotes mitotic rescue from transcription-mediated replication stress in SETX-deficient cancer cells

et al. 2022

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Replication stress (RS) is a leading cause of genome instability and cancer development. A substantial source of endogenous RS originates from the encounter between the transcription and replication machineries operating on the same DNA template. This occurs predominantly under specific contexts, such as oncogene activation, metabolic stress, or a deficiency in proteins that specifically act to prevent or resolve those transcription-replication conflicts (TRCs). One such protein is Senataxin (SETX), an RNA:DNA helicase involved in resolution of TRCs and R-loops. Here we identify a synthetic lethal interaction between SETX and proteins of the Fanconi anemia (FA) pathway. Depletion of SETX induces spontaneous under-replication and chromosome fragility due to active transcription and R-loops that persist in mitosis. These fragile loci are targeted by the Fanconi anemia protein, FANCD2, to facilitate the resolution of under-replicated DNA, thus preventing chromosome mis-segregation and allowing cells to proliferate. Mechanistically, we show that FANCD2 promotes mitotic DNA synthesis that is dependent on XPF and MUS81 endonucleases. Importantly, co-depleting FANCD2 together with SETX impairs cancer cell proliferation, without significantly affecting non-cancerous cells. Therefore, we uncovered a synthetic lethality between SETX and FA proteins for tolerance of transcription-mediated RS that may be exploited for cancer therapy.

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Section: Discussionmentioning

confidence: 99%

FANCD2 promotes mitotic rescue from transcription-mediated replication stress in SETX-deficient cancer cells

et al. 2022

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“…During this step, we determine the optimal concentration of CDK1 inhibitor (RO-3306) required to effectively arrest cells released from thymidine block in G2. To do this, we use FACS analyses of DNA content combined with immunofluorescent staining for phosphorylated Histone H3 (Ser10) 1 ( Figure 1 B), which is a marker for cells that entered mitosis. Seed 1.6 × 10 5 H1299 cells per well of a 6-well plate.…”

Section: Before You Beginmentioning

confidence: 99%

“…For complete details on the use and execution of this protocol, please refer to Groelly et al. (2022) 1 and Macheret et al. (2020).…”

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confidence: 99%

High-resolution mapping of mitotic DNA synthesis under conditions of replication stress in cultured cells

et al. 2023

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“…Recently, it has been reported that the frequency of MiDAS is elevated in BRCA2 deficient cells [ 157 ]. However, most of the EdU-sequenced regions in BRCA2-deficient mitotic cells did not overlap with APH-induced MiDAS.…”

Section: Mechanisms For Managing Replication Stressmentioning

confidence: 99%

“…These DNA lesions turnout to be the legacy of DNA repair in mitosis. This phenotype was restored upon overexpression of RNase H1 highlighting on the R-loops as a possible inducer of MiDAS [ 157 ].…”

Section: Mechanisms For Managing Replication Stressmentioning

confidence: 99%

Safeguarding DNA Replication: A Golden Touch of MiDAS and Other Mechanisms

Nachar

Rosselli

2022

IJMS

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DNA replication is a tightly regulated fundamental process allowing the correct duplication and transfer of the genetic information from the parental cell to the progeny. It involves the coordinated assembly of several proteins and protein complexes resulting in replication fork licensing, firing and progression. However, the DNA replication pathway is strewn with hurdles that affect replication fork progression during S phase. As a result, cells have adapted several mechanisms ensuring replication completion before entry into mitosis and segregating chromosomes with minimal, if any, abnormalities. In this review, we describe the possible obstacles that a replication fork might encounter and how the cell manages to protect DNA replication from S to the next G1.

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Mitotic DNA synthesis is caused by transcription-replication conflicts in BRCA2-deficient cells

Cited by 24 publications

References 79 publications

FANCD2 promotes mitotic rescue from transcription-mediated replication stress in SETX-deficient cancer cells

FANCD2 promotes mitotic rescue from transcription-mediated replication stress in SETX-deficient cancer cells

High-resolution mapping of mitotic DNA synthesis under conditions of replication stress in cultured cells

Safeguarding DNA Replication: A Golden Touch of MiDAS and Other Mechanisms

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