TOPOVIBL-REC114 interaction regulates meiotic DNA double-strand breaks

Meiosis requires the formation of programmed DNA double strand breaks (DSBs), essential for fertility and for generating genetic diversity. In male and female meiotic cells, DSBs are induced by the catalytic activity of the TOPOVIL complex formed by SPO11 and TOPOVIBL. To ensure genomic integrity, DNA cleavage activity is tightly regulated, and several accessory factors (REC114, MEI4, IHO1, and MEI1) are needed for DSB formation in mice. How and when these proteins act is not understood. Here, we show that REC114 is a direct partner of TOPOVIBL, and identified their conserved interacting domains by structural analysis. We then analysed the role of this interaction by monitoring meiotic DSBs in female and male mice carrying point mutations in TOPOVIBL that decrease or disrupt its binding to REC114. In these mutants, DSB activity was strongly reduced genome-wide in oocytes, but only in sub-telomeric regions in spermatocytes. In addition, in mutant spermatocytes, DSB activity was delayed in autosomes. These results provide evidence that REC114 is a key member of the TOPOVIL catalytic complex, and that the REC114/TOPOVIBL interaction ensures the efficiency and timing of DSB activity by integrating specific chromosomal features.

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De novo deletion mutations at recombination hotspots in mouse germlines

10.1101/2020.06.23.168138 ◽

2020 ◽

Author(s):

Agnieszka Lukaszewicz ◽

Julian Lange ◽

Scott Keeney ◽

Maria Jasin

Keyword(s):

De Novo ◽

Double Strand Breaks ◽

Dna Double Strand Breaks ◽

Atm Kinase ◽

Strand Breaks ◽

Homologous Chromosomes ◽

Recombination Hotspots ◽

Genome Wide ◽

Meiotic Dsbs ◽

Meiotic Recombination Hotspots

AbstractNumerous DNA double-strand breaks (DSBs) arise genome-wide during meiosis to ensure recombination between homologous chromosomes, which is required for gamete formation1,2. The ATM kinase plays a central role in controlling both the number and position of DSBs3-5, but the consequences of deregulated DSB formation have not been explored. Here we discovered that an unanticipated type of DNA deletion arises at meiotic recombination hotspots in the absence of ATM. Deletions form via joining of ends from two closely-spaced DSBs at adjacent hotspots or within a single hotspot. Deletions are also detected in normal cells, albeit at much lower frequency, revealing that the meiotic genome has a hidden potential for deletion events. Remarkably, a subset of deletions contain insertions that likely originated from DNA fragments released from hotspots on other chromosomes. Moreover, although deletions form primarily within one chromosome, joining between homologous chromosomes is also observed. This predicts in turn gross chromosome rearrangements, with evidence of damage to multiple chromatids and aborted gap repair. Thus, multiple nearby meiotic DSBs are normally suppressed by ATM to protect genomic integrity. We expect the de novo germline mutations we observe to affect human health and genome evolution.

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Extensive sex differences at the initiation of genetic recombination

10.1101/235002 ◽

2017 ◽

Author(s):

Kevin Brick ◽

Sarah Thibault-Sennett ◽

Fatima Smagulova ◽

Kwan-Wood G. Lam ◽

Yongmei Pu ◽

...

Keyword(s):

Sex Differences ◽

Chromosomal Abnormalities ◽

Genetic Recombination ◽

Double Strand Breaks ◽

Local Context ◽

Dna Double Strand Breaks ◽

Strand Breaks ◽

Genome Wide ◽

Meiotic Dsbs ◽

Initiation Stage

AbstractHomologous recombination in meiosis is initiated by programmed DNA double strand breaks (DSBs) and DSB repair as a crossover is essential to prevent chromosomal abnormalities in gametes. Sex differences in recombination have been previously observed by analyses of recombination end-products. To understand when and how sex differences are established, we built genome-wide maps of meiotic DSBs in both male and female mice. We found that most recombination initiates at sex-biased DSB hotspots. Local context, the choice of DSB targeting pathway and sex-specific patterns of DNA methylation give rise to these differences. Sex differences are not limited to the initiation stage, as the rate at which DSBs are repaired as crossovers appears to differ between the sexes in distal regions. This uneven repair patterning may be linked to the higher aneuploidy rate in females. Together, these data demonstrate that sex differences occur early in meiotic recombination.

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Faculty Opinions recommendation of DNA double-strand breaks trigger genome-wide sister-chromatid cohesion through Eco1 (Ctf7).

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1090512.545649 ◽

2007 ◽

Author(s):

Kerry Bloom

Keyword(s):

Sister Chromatid ◽

Sister Chromatid Cohesion ◽

Double Strand Breaks ◽

Dna Double Strand Breaks ◽

Strand Breaks ◽

Genome Wide ◽

Chromatid Cohesion

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Faculty Opinions recommendation of DNA double-strand breaks trigger genome-wide sister-chromatid cohesion through Eco1 (Ctf7).

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1090512.543783 ◽

2007 ◽

Author(s):

Antony Carr

Keyword(s):

Sister Chromatid ◽

Sister Chromatid Cohesion ◽

Double Strand Breaks ◽

Dna Double Strand Breaks ◽

Strand Breaks ◽

Genome Wide ◽

Chromatid Cohesion

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Genome-wide mapping of DNA double-strand breaks from eukaryotic cell cultures using Break-seq

STAR Protocols ◽

10.1016/j.xpro.2021.100554 ◽

2021 ◽

Vol 2 (2) ◽

pp. 100554

Author(s):

Ishita Joshi ◽

Jenna DeRycke ◽

Megan Palmowski ◽

Robert LeSuer ◽

Wenyi Feng

Keyword(s):

Cell Cultures ◽

Eukaryotic Cell ◽

Double Strand Breaks ◽

Dna Double Strand Breaks ◽

Strand Breaks ◽

Genome Wide

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i-BLESS is an ultra-sensitive method for detection of DNA double-strand breaks

Communications Biology ◽

10.1038/s42003-018-0165-9 ◽

2018 ◽

Vol 1 (1) ◽

Cited By ~ 22

Author(s):

Anna Biernacka ◽

Yingjie Zhu ◽

Magdalena Skrzypczak ◽

Romain Forey ◽

Benjamin Pardo ◽

...

Keyword(s):

Cell Fate ◽

Genome Stability ◽

Strand Break ◽

Double Strand Breaks ◽

Universal Method ◽

Dna Double Strand Breaks ◽

Strand Breaks ◽

Agarose Beads ◽

Genome Wide ◽

Dna Double Strand Break

AbstractMaintenance of genome stability is a key issue for cell fate that could be compromised by chromosome deletions and translocations caused by DNA double-strand breaks (DSBs). Thus development of precise and sensitive tools for DSBs labeling is of great importance for understanding mechanisms of DSB formation, their sensing and repair. Until now there has been no high resolution and specific DSB detection technique that would be applicable to any cells regardless of their size. Here, we present i-BLESS, a universal method for direct genome-wide DNA double-strand break labeling in cells immobilized in agarose beads. i-BLESS has three key advantages: it is the only unbiased method applicable to yeast, achieves a sensitivity of one break at a given position in 100,000 cells, and eliminates background noise while still allowing for fixation of samples. The method allows detection of ultra-rare breaks such as those forming spontaneously at G-quadruplexes.

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Genetic Separation of Sae2 Nuclease Activity from Mre11 Nuclease Functions in Budding Yeast

Molecular and Cellular Biology ◽

10.1128/mcb.00156-17 ◽

2017 ◽

Vol 37 (24) ◽

Cited By ~ 8

Author(s):

Sucheta Arora ◽

Rajashree A. Deshpande ◽

Martin Budd ◽

Judy Campbell ◽

America Revere ◽

...

Keyword(s):

Nuclease Activity ◽

Double Strand Breaks ◽

Endonuclease Activity ◽

Dna Double Strand Breaks ◽

Dna Breaks ◽

Content Type ◽

Strand Breaks ◽

Dna Ends

ABSTRACT Sae2 promotes the repair of DNA double-strand breaks in Saccharomyces cerevisiae. The role of Sae2 is linked to the Mre11/Rad50/Xrs2 (MRX) complex, which is important for the processing of DNA ends into single-stranded substrates for homologous recombination. Sae2 has intrinsic endonuclease activity, but the role of this activity has not been assessed independently from its functions in promoting Mre11 nuclease activity. Here we identify and characterize separation-of-function mutants that lack intrinsic nuclease activity or the ability to promote Mre11 endonucleolytic activity. We find that the ability of Sae2 to promote MRX nuclease functions is important for DNA damage survival, particularly in the absence of Dna2 nuclease activity. In contrast, Sae2 nuclease activity is essential for DNA repair when the Mre11 nuclease is compromised. Resection of DNA breaks is impaired when either Sae2 activity is blocked, suggesting roles for both Mre11 and Sae2 nuclease activities in promoting the processing of DNA ends in vivo. Finally, both activities of Sae2 are important for sporulation, indicating that the processing of meiotic breaks requires both Mre11 and Sae2 nuclease activities.

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CTCF cooperates with CtIP to drive homologous recombination repair of double-strand breaks

Nucleic Acids Research ◽

10.1093/nar/gkz639 ◽

2019 ◽

Vol 47 (17) ◽

pp. 9160-9179 ◽

Cited By ~ 6

Author(s):

Soon Young Hwang ◽

Mi Ae Kang ◽

Chul Joon Baik ◽

Yejin Lee ◽

Ngo Thanh Hang ◽

...

Keyword(s):

Homologous Recombination ◽

Critical Role ◽

Control Point ◽

Double Strand Breaks ◽

Dna Double Strand Breaks ◽

Strand Breaks ◽

C Terminus ◽

Dna End Resection ◽

End Resection

Abstract The pleiotropic CCCTC-binding factor (CTCF) plays a role in homologous recombination (HR) repair of DNA double-strand breaks (DSBs). However, the precise mechanistic role of CTCF in HR remains largely unclear. Here, we show that CTCF engages in DNA end resection, which is the initial, crucial step in HR, through its interactions with MRE11 and CtIP. Depletion of CTCF profoundly impairs HR and attenuates CtIP recruitment at DSBs. CTCF physically interacts with MRE11 and CtIP and promotes CtIP recruitment to sites of DNA damage. Subsequently, CTCF facilitates DNA end resection to allow HR, in conjunction with MRE11–CtIP. Notably, the zinc finger domain of CTCF binds to both MRE11 and CtIP and enables proficient CtIP recruitment, DNA end resection and HR. The N-terminus of CTCF is able to bind to only MRE11 and its C-terminus is incapable of binding to MRE11 and CtIP, thereby resulting in compromised CtIP recruitment, DSB resection and HR. Overall, this suggests an important function of CTCF in DNA end resection through the recruitment of CtIP at DSBs. Collectively, our findings identify a critical role of CTCF at the first control point in selecting the HR repair pathway.

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