scholarly journals Signaling to TRP53 and TAp63 from CHK1/CHK2 is responsible for elimination of most oocytes defective for either chromosome synapsis or recombination

2019 ◽  
Author(s):  
Vera D. Rinaldi ◽  
Jordana C. Bloom ◽  
John C. Schimenti
Keyword(s):  
Threshold Level ◽  
Double Strand Breaks ◽  
Checkpoint Kinase ◽  
Strand Breaks ◽  
Homologous Chromosomes ◽  
Chromosome Synapsis ◽  
Checkpoint Pathway ◽  
Mutation Avoidance ◽  
Meiotic Dsbs ◽  
Eukaryotic Organisms

ABSTRACTEukaryotic organisms have evolved mechanisms to prevent the accumulation of cells bearing genetic aberrations. This is especially crucial for the germline, because fecundity, and fitness of progeny would be adversely affected by an excessively high mutational incidence. The process of meiosis poses unique problems for mutation avoidance, due to the requirement for SPO11-induced programmed double strand breaks (DSBs) in recombination-driven pairing and segregation of homologous chromosomes. Mouse meiocytes bearing unrepaired meiotic DSBs or unsynapsed chromosomes are eliminated before completing meiotic prophase I. In previous work, we showed that checkpoint kinase 2 (CHK2; CHEK2), a canonical DNA damage response protein, is crucial for eliminating not only oocytes defective in meiotic DSB repair (e.g. Trip13Gt mutants), but also asynaptic Spo11−/− oocytes that accumulate a threshold level of spontaneous DSBs. However, rescue of such oocytes by Chk2 deficiency was incomplete, raising the possibility that a parallel checkpoint pathway(s) exists. Here, we show that mouse oocytes lacking both TAp63 and TRP53 protects nearly all Spo11−/− and Trip13Gt/Gt oocytes from elimination. We present evidence that checkpoint kinase I (CHK1; CHEK1), which is known to signal to TRP53, also becomes activated by persistent DSBs in oocytes, and to an increased degree when CHK2 is absent. The combined data indicate that nearly all oocytes reaching a threshold level of unrepaired DSBs are eliminated by a semi-redundant pathway of CHK1/CHK2 signaling to TRP53/TAp63.

scholarly journals Oocyte Elimination Through DNA Damage Signaling from CHK1/CHK2 to p53 and p63

Genetics ◽  
2020 ◽  
Vol 215 (2) ◽  
pp. 373-378 ◽  
Author(s):  
Vera D. Rinaldi ◽  
Jordana C. Bloom ◽  
John C. Schimenti
Keyword(s):  
Dna Damage ◽  
Threshold Level ◽  
Checkpoint Kinase ◽  
Strand Breaks ◽  
Dna Damage Signaling ◽  
Chromosome Synapsis ◽  
Checkpoint Pathway ◽  
Mutation Avoidance ◽  
Meiotic Dsbs ◽  
Eukaryotic Organisms

Eukaryotic organisms have evolved mechanisms to prevent the accumulation of cells bearing genetic aberrations. This is especially crucial for the germline, because fecundity and fitness of progeny would be adversely affected by an excessively high mutational incidence. The process of meiosis poses unique problems for mutation avoidance because of the requirement for SPO11-induced programmed double-strand breaks (DSBs) in recombination-driven pairing and segregation of homologous chromosomes. Mouse meiocytes bearing unrepaired meiotic DSBs or unsynapsed chromosomes are eliminated before completing meiotic prophase I. In previous work, we showed that checkpoint kinase 2 (CHK2; CHEK2), a canonical DNA damage response protein, is crucial for eliminating not only oocytes defective in meiotic DSB repair (e.g., Trip13Gt mutants), but also Spo11−/− oocytes that are defective in homologous chromosome synapsis and accumulate a threshold level of spontaneous DSBs. However, rescue of such oocytes by Chk2 deficiency was incomplete, raising the possibility that a parallel checkpoint pathway(s) exists. Here, we show that mouse oocytes lacking both p53 (TRP53) and the oocyte-exclusive isoform of p63, TAp63, protects nearly all Spo11−/− and Trip13Gt/Gt oocytes from elimination. We present evidence that checkpoint kinase I (CHK1; CHEK1), which is known to signal to TRP53, also becomes activated by persistent DSBs in oocytes, and to an increased degree when CHK2 is absent. The combined data indicate that nearly all oocytes reaching a threshold level of unrepaired DSBs are eliminated by a semiredundant pathway of CHK1/CHK2 signaling to TRP53/TAp63.

scholarly journals Tex19.1 Regulates Meiotic DNA Double Strand Break Frequency in Mouse Spermatocytes

2017 ◽  
Author(s):  
James H. Crichton ◽  
Christopher J. Playfoot ◽  
Marie MacLennan ◽  
David Read ◽  
Howard J. Cooke ◽  
...  
Keyword(s):  
Ubiquitin Ligase ◽  
Meiotic Recombination ◽  
Homologous Chromosome ◽  
E3 Ubiquitin Ligase ◽  
Double Strand Breaks ◽  
Dna Double Strand Breaks ◽  
Genetic Pathway ◽  
Strand Breaks ◽  
Chromosome Synapsis ◽  
Meiotic Dsbs

AbstractMeiosis relies on the SPO11 endonuclease to generate the recombinogenic DNA double strand breaks (DSBs) required for homologous chromosome synapsis and segregation. The number of meiotic DSBs needs to be sufficient to allow chromosomes to search for and find their homologs, but not excessive to the point of causing genome instability. Here we report that meiotic DSB frequency in mouse spermatocytes is regulated by the mammal-specific gene Tex19.1. We show that the chromosome asynapsis previously reported in Tex19.1-/- spermatocytes is preceded by reduced numbers of recombination foci in leptotene and zygotene. Tex19.1 is required for the generation of normal levels of Spo11-dependent DNA damage during leptotene, but not for upstream events such as MEI4 foci formation or accumulation of H3K4me3 at recombination hotspots. Furthermore, we show that mice carrying mutations in the E3 ubiquitin ligase UBR2, a TEX19.1-interacting partner, phenocopy the Tex19.1-/- recombination defects. These data show that Tex19.1 and Ubr2 are required for mouse spermatocytes to generate sufficient meiotic DSBs to ensure that homology search is consistently successful, and reveal a hitherto unknown genetic pathway regulating meiotic DSB frequency in mammals.Author SummaryMeiosis is a specialised type of cell division that occurs during sperm and egg development to reduce chromosome number prior to fertilisation. Recombination is a key step in meiosis as it facilitates the pairing of homologous chromosomes prior to their reductional division, and generates new combinations of genetic alleles for transmission in the next generation. Regulating the amount of recombination is key for successful meiosis: too much will likely cause mutations, chromosomal re-arrangements and genetic instability, whereas too little causes defects in homologous chromosome pairing prior to the meiotic divisions. This study identifies a genetic pathway requiredto generate robust meiotic recombination in mouse spermatocytes. We show that male mice with mutations in Tex19.1 or Ubr2, which encodes an E3 ubiquitin ligase that interacts with TEX19.1, have defects in generating normal levels of meiotic recombination. We show that the defects in these mutants impact on the recombination process at the stage when programmed DNA double strand breaks are being made. This defect likely contributes to the chromosome synapsis and meiotic progression phenotypes previously described in these mutant mice. This study has implications for our understanding of how this fundamental aspect of genetics and inheritance is controlled.

scholarly journals Massive crossover elevation via combination of HEI10 and recq4a recq4b during Arabidopsis meiosis

2018 ◽  
Vol 115 (10) ◽  
pp. 2437-2442 ◽  
Author(s):  
Heïdi Serra ◽  
Christophe Lambing ◽  
Catherine H. Griffin ◽  
Stephanie D. Topp ◽  
Divyashree C. Nageswaran ◽  
...  
Keyword(s):  
Meiotic Recombination ◽  
Homologous Chromosome ◽  
Crop Improvement ◽  
Double Strand Breaks ◽  
Dna Double Strand Breaks ◽  
Dsb Repair ◽  
Strand Breaks ◽  
Homologous Chromosomes ◽  
Meiotic Dsbs ◽  
Competing Pathways

During meiosis, homologous chromosomes undergo reciprocal crossovers, which generate genetic diversity and underpin classical crop improvement. Meiotic recombination initiates from DNA double-strand breaks (DSBs), which are processed into single-stranded DNA that can invade a homologous chromosome. The resulting joint molecules can ultimately be resolved as crossovers. In Arabidopsis, competing pathways balance the repair of ∼100–200 meiotic DSBs into ∼10 crossovers per meiosis, with the excess DSBs repaired as noncrossovers. To bias DSB repair toward crossovers, we simultaneously increased dosage of the procrossover E3 ligase gene HEI10 and introduced mutations in the anticrossovers helicase genes RECQ4A and RECQ4B. As HEI10 and recq4a recq4b increase interfering and noninterfering crossover pathways, respectively, they combine additively to yield a massive meiotic recombination increase. Interestingly, we also show that increased HEI10 dosage increases crossover coincidence, which indicates an effect on interference. We also show that patterns of interhomolog polymorphism and heterochromatin drive recombination increases distally towards the subtelomeres in both HEI10 and recq4a recq4b backgrounds, while the centromeres remain crossover suppressed. These results provide a genetic framework for engineering meiotic recombination landscapes in plant genomes.

scholarly journals Massive crossover elevation via combination of HEI10 and recq4a recq4b during Arabidopsis meiosis

2017 ◽  
Author(s):  
Heïdi Serra ◽  
Christophe Lambing ◽  
Catherine H. Griffin ◽  
Stephanie D. Topp ◽  
Mathilde Séguéla-Arnaud ◽  
...  
Keyword(s):  
Meiotic Recombination ◽  
Homologous Chromosome ◽  
Crop Improvement ◽  
Double Strand Breaks ◽  
Dna Double Strand Breaks ◽  
Dsb Repair ◽  
Strand Breaks ◽  
Homologous Chromosomes ◽  
Meiotic Dsbs ◽  
Competing Pathways

AbstractDuring meiosis homologous chromosomes undergo reciprocal crossovers, which generate genetic diversity and underpin classical crop improvement. Meiotic recombination initiates from DNA double strand breaks, which are processed into single-stranded DNA that can invade a homologous chromosome. The resulting joint molecules can ultimately be resolved as crossovers. In Arabidopsis, competing pathways balance the repair of ∼100–200 meiotic DSBs into ∼10 crossovers per meiosis, with the excess DSBs repaired as non-crossovers. In order to bias DSB repair towards crossovers, we simultaneously increased dosage of the pro-crossover E3 ligase gene HEI10 and introduced mutations in the anti-crossover helicase genes RECQ4A and RECQ4B. As HEI10 and recq4a recq4b increase interfering and non-interfering crossover pathways respectively, they combine additively to yield a massive meiotic recombination increase. Interestingly, we also show that increased HEI10 dosage increases crossover coincidence, which indicates an effect of HEI10 on interference. We also show that patterns of interhomolog polymorphism and heterochromatin drive recombination increases towards the sub-telomeres in both HEI10 and recq4a recq4b backgrounds, while the centromeres remain crossover-suppressed. These results provide a genetic framework for engineering meiotic recombination landscapes in plant genomes.

scholarly journals A single checkpoint pathway eliminates mouse oocytes with dna damage or chromosome synapsis failure

2017 ◽  
Author(s):  
Vera D. Rinaldi ◽  
Ewelina Bolcun-Filas ◽  
Hiroshi Kogo ◽  
Hiroki Kurahashi ◽  
John C. Schimenti
Keyword(s):  
Dna Damage ◽  
Substantial Fraction ◽  
Activation Threshold ◽  
Late Prophase ◽  
Checkpoint Activation ◽  
Homologous Chromosomes ◽  
Mouse Oocytes ◽  
Chromosome Synapsis ◽  
Checkpoint Pathway ◽  
Meiotic Dsbs

Pairing and synapsis of homologous chromosomes during meiosis is crucial for producing genetically normal gametes, and is dependent upon repair of SPO11-induced double stranded breaks (DSBs) by homologous recombination. To prevent transmission of genetic defects, diverse organisms have evolved mechanisms to eliminate meiocytes containing unrepaired DSBs or unsynapsed chromosomes. Here, we show that the CHK2 (CHEK2)-dependent DNA damage checkpoint culls not only recombination-defective mouse oocytes, but also SPO11-deficient oocytes that are severely defective in homolog synapsis. The checkpoint is triggered by spontaneous DSBs that arise in late prophase I, accumulating above the checkpoint activation threshold (∼10 DSBs) because presence of HORMAD1/2 on unsynapsed chromosome axes prevents their repair. Furthermore, Hormad2 deletion rescued fertility and meiotic DSB repair of oocytes containing a synapsis-proficient, non-crossover recombination defective mutation in a gene (Trip13) required for removal of HORMADs from synapsed chromosomes, indicating that a substantial fraction of meiotic DSBs are normally repaired by intersister recombination in mice.

scholarly journals Three Distinct Modes of Mec1/ATR and Tel1/ATM Activation Illustrate Differential Checkpoint Targeting during Budding Yeast Early Meiosis

2013 ◽  
Vol 33 (16) ◽  
pp. 3365-3376 ◽  
Author(s):  
Yun-Hsin Cheng ◽  
Chi-Ning Chuang ◽  
Hui-Ju Shen ◽  
Feng-Ming Lin ◽  
Ting-Fang Wang
Keyword(s):  
Dna Replication ◽  
Budding Yeast ◽  
Noncovalent Interactions ◽  
Double Strand Breaks ◽  
Histone H2a ◽  
Dna Double Strand Breaks ◽  
Dsb Repair ◽  
Strand Breaks ◽  
Homologous Chromosomes ◽  
Chromosome Synapsis

Recombination and synapsis of homologous chromosomes are hallmarks of meiosis in many organisms. Meiotic recombination is initiated by Spo11-induced DNA double-strand breaks (DSBs), whereas chromosome synapsis is mediated by a tripartite structure named the synaptonemal complex (SC). Previously, we proposed that budding yeast SC is assembled via noncovalent interactions between the axial SC protein Red1, SUMO chains or conjugates, and the central SC protein Zip1. Incomplete synapsis and unrepaired DNA are monitored by Mec1/Tel1-dependent checkpoint responses that prevent exit from the pachytene stage. Here, our results distinguished three distinct modes of Mec1/Tec1 activation during early meiosis that led to phosphorylation of three targets, histone H2A at S129 (γH2A), Hop1, and Zip1, which are involved, respectively, in DNA replication, the interhomolog recombination and chromosome synapsis checkpoint, and destabilization of homology-independent centromere pairing. γH2A phosphorylation is Red1 independent and occurs prior to Spo11-induced DSBs. DSB- and Red1-dependent Hop1 phosphorylation is activated via interaction of the Red1-SUMO chain/conjugate ensemble with the Ddc1-Rad17-Mec3 (9-1-1) checkpoint complex and the Mre11-Rad50-Xrs2 complex. During SC assembly, Zip1 outcompetes 9-1-1 from the Red1-SUMO chain ensemble to attenuate Hop1 phosphorylation. In contrast, chromosome synapsis cannot attenuate DSB-dependent and Red1-independent Zip1 phosphorylation. These results reveal how DNA replication, DSB repair, and chromosome synapsis are differentially monitored by the meiotic checkpoint network.

scholarly journals De novo deletion mutations at recombination hotspots in mouse germlines

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.

scholarly journals Impeding DNA Break Repair Enables Oocyte Quality Control

2018 ◽  
Author(s):  
Huanyu Qiao ◽  
H.B.D. Prasada Rao ◽  
Yan Yun ◽  
Sumit Sandhu ◽  
Jared H. Fong ◽  
...  
Keyword(s):  
Quality Control ◽  
Oocyte Quality ◽  
Double Strand Breaks ◽  
Negative Regulator ◽  
Dna Double Strand Breaks ◽  
Strand Breaks ◽  
Homologous Chromosomes ◽  
Chromosome Synapsis ◽  
Sumo Ligase ◽  
Induced Apoptosis

SUMMARYOocyte quality control culls eggs with defects in meiosis. In mouse, oocyte death is triggered by defects in chromosome synapsis and recombination, which involve repair of programmed DNA double-strand breaks (DSBs) between homologous chromosomes. We show that RNF212, a SUMO ligase required for crossing over, also mediates oocyte quality control. Both physiological apoptosis and wholesale oocytes elimination in meiotic mutants require RNF212. RNF212 sensitizes cells to DSB-induced apoptosis within a narrow window when chromosomes desynapse during the transition into quiescence. Analysis of DNA damage during this transition implies that RNF212 impedes DSB repair. Consistently, RNF212 is required for HORMAD1, a negative regulator of inter-sister recombination, to associate with desynapsing chromosomes. We infer that oocytes impede repair of residual DSBs to retain a “memory” of meiotic defects that enables quality control processes. These results define the logic of oocyte quality control and suggest RNF212 variants may influence transmission of defective genomes.

scholarly journals Four-pronged negative feedback of DSB machinery in meiotic DNA-break control in mice

2021 ◽  
Author(s):  
Ihsan Dereli ◽  
Marcello Stanzione ◽  
Fabrizio Olmeda ◽  
Frantzeskos Papanikos ◽  
Marek Baumann ◽  
...  
Keyword(s):  
Negative Feedback ◽  
Protein Complexes ◽  
Double Strand Breaks ◽  
Dna Double Strand Breaks ◽  
Strand Breaks ◽  
Homologous Chromosomes ◽  
Damage Response ◽  
Auxiliary Protein ◽  
Genomic Locations ◽  
Spatiotemporal Control

Abstract In most taxa, halving of chromosome numbers during meiosis requires that homologous chromosomes (homologues) pair and form crossovers. Crossovers emerge from the recombination-mediated repair of programmed DNA double-strand breaks (DSBs). DSBs are generated by SPO11, whose activity requires auxiliary protein complexes, called pre-DSB recombinosomes. To elucidate the spatiotemporal control of the DSB machinery, we focused on an essential SPO11 auxiliary protein, IHO1, which serves as the main anchor for pre-DSB recombinosomes on chromosome cores, called axes. We discovered that DSBs restrict the DSB machinery by at least four distinct pathways in mice. Firstly, by activating the DNA damage response (DDR) kinase ATM, DSBs restrict pre-DSB recombinosome numbers without affecting IHO1. Secondly, in their vicinity, DSBs trigger IHO1 depletion mainly by another DDR kinase, ATR. Thirdly, DSBs enable homologue synapsis, which promotes the depletion of IHO1 and pre-DSB recombinosomes from synapsed axes. Finally, DSBs and three DDR kinases, ATM, ATR and PRKDC, enable stage-specific depletion of IHO1 from all axes. We hypothesize that these four negative feedback pathways protect genome integrity by ensuring that DSBs form without excess, are well-distributed, and are restricted to genomic locations and prophase stages where DSBs are functional for promoting homologue pairing and crossover formation.

scholarly journals End-resection at DNA double-strand breaks in the three domains of life

2013 ◽  
Vol 41 (1) ◽  
pp. 314-320 ◽  
Author(s):  
John K. Blackwood ◽  
Neil J. Rzechorzek ◽  
Sian M. Bray ◽  
Joseph D. Maman ◽  
Luca Pellegrini ◽  
...  
Keyword(s):  
Dna Repair ◽  
Homologous Recombination ◽  
Strand Break ◽  
Double Strand Breaks ◽  
Dna Double Strand Breaks ◽  
Strand Breaks ◽  
Homologous Chromosomes ◽  
Domains Of Life ◽  
Strand Invasion ◽  
End Resection

During DNA repair by HR (homologous recombination), the ends of a DNA DSB (double-strand break) must be resected to generate single-stranded tails, which are required for strand invasion and exchange with homologous chromosomes. This 5′–3′ end-resection of the DNA duplex is an essential process, conserved across all three domains of life: the bacteria, eukaryota and archaea. In the present review, we examine the numerous and redundant helicase and nuclease systems that function as the enzymatic analogues for this crucial process in the three major phylogenetic divisions.

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