SPO16 binds SHOC1 to promote homologous recombination and crossing-over in meiotic prophase I

Segregation of homologous chromosomes in meiosis I is tightly regulated by their physical links, or crossovers (COs), generated from DNA double-strand breaks (DSBs) through meiotic homologous recombination. In budding yeast, three ZMM (Zip1/2/3/4, Mer3, Msh4/5) proteins, Zip2, Zip4, and Spo16, form a “ZZS” complex, functioning to promote meiotic recombination via a DSB repair pathway. Here, we identified the mammalian ortholog of Spo16, termed SPO16, which interacts with the mammalian ortholog of Zip2 (SHOC1/MZIP2), and whose functions are evolutionarily conserved to promote the formation of COs. SPO16 localizes to the recombination nodules, as SHOC1 and TEX11 do. SPO16 is required for stabilization of SHOC1 and proper localization of other ZMM proteins. The DSBs formed in SPO16-deleted meiocytes were repaired without COs formation, although synapsis is less affected. Therefore, formation of SPO16-SHOC1 complex–associated recombination intermediates is a key step facilitating meiotic recombination that produces COs from yeast to mammals.

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Double-strand-break-induced homologous recombination in mammalian cells

Biochemical Society Transactions ◽

10.1042/bst0290196 ◽

2001 ◽

Vol 29 (2) ◽

pp. 196-201 ◽

Cited By ~ 186

Author(s):

R. D. Johnson ◽

M. Jasin

Keyword(s):

Homologous Recombination ◽

Sister Chromatid ◽

Mammalian Cells ◽

Indirect Evidence ◽

Strand Break ◽

Double Strand Breaks ◽

Crossing Over ◽

Dna Double Strand Breaks ◽

Dsb Repair ◽

Strand Breaks

In mammalian cells, the repair of DNA double-strand breaks (DSBs) occurs by both homologous and non-homologous mechanisms. Indirect evidence, including that from gene targeting and random integration experiments, had suggested that non-homologous mechanisms were significantly more frequent than homologous ones. However, more recent experiments indicate that homologous recombination is also a prominent DSB repair pathway. These experiments show that mammalian cells use homologous sequences located at multiple positions throughout the genome to repair a DSB. However, template preference appears to be biased, with the sister chromatid being preferred by 2–3 orders of magnitude over a homologous or heterologous chromosome. The outcome of homologous recombination in mammalian cells is predominantly gene conversion that is not associated with crossing-over. The preference for the sister chromatid and the bias against crossing-over seen in mitotic mammalian cells may have developed in order to reduce the potential for genome alterations that could occur when other homologous repair templates are utilized. In attempts to understand further the mechanism of homologous recombination, the proteins that promote this process are beginning to be identified. To date, four mammalian proteins have been demonstrated conclusively to be involved in DSB repair by homologous recombination: Rad54, XRCC2, XRCC3 and BRCAI. This paper summarizes results from a number of recent studies.

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

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1713071115 ◽

2018 ◽

Vol 115 (10) ◽

pp. 2437-2442 ◽

Cited By ~ 38

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.

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

10.1101/159764 ◽

2017 ◽

Cited By ~ 2

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.

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Coupling crossover and synaptonemal complex in meiosis

Genes & Development ◽

10.1101/gad.349286.121 ◽

2022 ◽

Vol 36 (1-2) ◽

pp. 4-6

Author(s):

Corinne Grey ◽

Bernard de Massy

Keyword(s):

Saccharomyces Cerevisiae ◽

Homologous Recombination ◽

Synaptonemal Complex ◽

Double Strand Breaks ◽

Dna Double Strand Breaks ◽

Dsb Repair ◽

Strand Breaks ◽

Homologous Chromosomes

During meiosis, a molecular program induces DNA double-strand breaks (DSBs) and their repair by homologous recombination. DSBs can be repaired with or without crossovers. ZMM proteins promote the repair toward crossover. The sites of DSB repair are also sites where the axes of homologous chromosomes are juxtaposed and stabilized, and where a structure called the synaptonemal complex initiates, providing further regulation of both DSB formation and repair. How crossover formation and synapsis initiation are linked has remained unknown. The study by Pyatnitskaya and colleagues (pp. 53–69) in this issue of Genes & Development highlights the central role of the Saccharomyces cerevisiae ZMM protein Zip4 in this process.

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MEIOK21: a new component of meiotic recombination bridges required for spermatogenesis

Nucleic Acids Research ◽

10.1093/nar/gkaa406 ◽

2020 ◽

Vol 48 (12) ◽

pp. 6624-6639

Author(s):

Yongliang Shang ◽

Tao Huang ◽

Hongbin Liu ◽

Yanlei Liu ◽

Heng Liang ◽

...

Keyword(s):

Meiotic Recombination ◽

Double Strand Breaks ◽

Strand Exchange ◽

Homology Search ◽

Physical Interaction ◽

Dna Double Strand Breaks ◽

Dsb Repair ◽

Chromosome Localization ◽

Strand Breaks ◽

Homologous Chromosomes

Abstract Repair of DNA double-strand breaks (DSBs) with homologous chromosomes is a hallmark of meiosis that is mediated by recombination ‘bridges’ between homolog axes. This process requires cooperation of DMC1 and RAD51 to promote homology search and strand exchange. The mechanism(s) regulating DMC1/RAD51-ssDNA nucleoprotein filament and the components of ‘bridges’ remain to be investigated. Here we show that MEIOK21 is a newly identified component of meiotic recombination bridges and is required for efficient formation of DMC1/RAD51 foci. MEIOK21 dynamically localizes on chromosomes from on-axis foci to ‘hanging foci’, then to ‘bridges’, and finally to ‘fused foci’ between homolog axes. Its chromosome localization depends on DSBs. Knockout of Meiok21 decreases the numbers of HSF2BP and DMC1/RAD51 foci, disrupting DSB repair, synapsis and crossover recombination and finally causing male infertility. Therefore, MEIOK21 is a novel recombination factor and probably mediates DMC1/RAD51 recruitment to ssDNA or their stability on chromosomes through physical interaction with HSF2BP.

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

Biochemical Society Transactions ◽

10.1042/bst20120307 ◽

2013 ◽

Vol 41 (1) ◽

pp. 314-320 ◽

Cited By ~ 30

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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RAD51 Is Required for the Repair of Plasmid Double-Stranded DNA Gaps from Either Plasmid or Chromosomal Templates

Molecular and Cellular Biology ◽

10.1128/mcb.20.4.1194-1205.2000 ◽

2000 ◽

Vol 20 (4) ◽

pp. 1194-1205 ◽

Cited By ~ 63

Author(s):

Stephan Bärtsch ◽

Leslie E. Kang ◽

Lorraine S. Symington

Keyword(s):

Homologous Recombination ◽

Double Strand Breaks ◽

Crossing Over ◽

Dna Double Strand Breaks ◽

End Joining ◽

Radiation Chemical ◽

Strand Breaks ◽

Independent Events ◽

In The Wild ◽

A Minor

ABSTRACT DNA double-strand breaks may be induced by endonucleases, ionizing radiation, chemical agents, and mechanical forces or by replication of single-stranded nicked chromosomes. Repair of double-strand breaks can occur by homologous recombination or by nonhomologous end joining. A system was developed to measure the efficiency of plasmid gap repair by homologous recombination using either chromosomal or plasmid templates. Gap repair was biased toward gene conversion events unassociated with crossing over using either donor sequence. The dependence of recombinational gap repair on genes belonging to the RAD52epistasis group was tested in this system. RAD51,RAD52, RAD57, and RAD59 were required for efficient gap repair using either chromosomal or plasmid donors. No homologous recombination products were recovered fromrad52 mutants, whereas a low level of repair occurred in the absence of RAD51, RAD57, orRAD59. These results suggest a minor pathway of strand invasion that is dependent on RAD52 but not onRAD51. The residual repair events in rad51mutants were more frequently associated with crossing over than was observed in the wild-type strain, suggesting that the mechanisms forRAD51-dependent and RAD51-independent events are different. Plasmid gap repair was reduced synergistically inrad51 rad59 double mutants, indicating an important role for RAD59 in RAD51-independent repair.

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Vilya, a component of the recombination nodule, is required for meiotic double-strand break formation in Drosophila

eLife ◽

10.7554/elife.08287 ◽

2015 ◽

Vol 4 ◽

Cited By ~ 23

Author(s):

Cathleen M Lake ◽

Rachel J Nielsen ◽

Fengli Guo ◽

Jay R Unruh ◽

Brian D Slaughter ◽

...

Keyword(s):

Meiotic Recombination ◽

Strand Break ◽

Double Strand Breaks ◽

Crossing Over ◽

Actual Process ◽

Strand Breaks ◽

Recombination Nodules ◽

Immuno Electron Microscopy ◽

Break Formation ◽

Selection Of

Meiotic recombination begins with the induction of programmed double-strand breaks (DSBs). In most organisms only a fraction of DSBs become crossovers. Here we report a novel meiotic gene, vilya, which encodes a protein with homology to Zip3-like proteins shown to determine DSB fate in other organisms. Vilya is required for meiotic DSB formation, perhaps as a consequence of its interaction with the DSB accessory protein Mei-P22, and localizes to those DSB sites that will mature into crossovers. In early pachytene Vilya localizes along the central region of the synaptonemal complex and to discrete foci. The accumulation of Vilya at foci is dependent on DSB formation. Immuno-electron microscopy demonstrates that Vilya is a component of recombination nodules, which mark the sites of crossover formation. Thus Vilya links the mechanism of DSB formation to either the selection of those DSBs that will become crossovers or to the actual process of crossing over.

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A pathway for error-free non-homologous end joining of resected meiotic double-strand breaks

10.1101/2020.06.17.157313 ◽

2020 ◽

Author(s):

Talia Hatkevich ◽

Danny E. Miller ◽

Carolyn A. Turcotte ◽

Margaret C. Miller ◽

Jeff Sekelsky

Keyword(s):

Meiotic Recombination ◽

Homologous Chromosome ◽

Double Strand Breaks ◽

Dna Double Strand Breaks ◽

Repair Mechanism ◽

End Joining ◽

Dsb Repair ◽

Strand Breaks ◽

Recombination Processes ◽

Non Homologous End Joining

ABSTRACTProgrammed DNA double-strand breaks (DSBs) made during meiosis are repaired by recombination with the homologous chromosome to generate, at selected sites, reciprocal crossovers that are critical for the proper separation of homologs in the first meiotic divisions. Backup repair processes can compensate when the normal meiotic recombination processes are non-functional. We describe a novel backup repair mechanism that occurs when the homologous chromosome is not available in Drosophila melanogaster meiosis. In the presence of a previously described mutation (Mcm5A7) that disrupts chromosome pairing, DSB repair is initiated by homologous recombination but is completed by non-homologous end joining (NHEJ). Remarkably, this process yields precise repair products. Our results provide support for a recombination intermediate recently discovered in mouse meiosis, in which an oligonucleotide bound to the Spo11 protein that catalyzes DSB formation remains bound after resection. We propose that this oligonucleotide functions as a primer for fill-in synthesis to allow scarless repair by NHEJ.

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CRISPR targeting of MEIOTIC-TOPOISOMERASE VIB-dCas9 to a recombination hotspot is insufficient to increase crossover frequency in Arabidopsis

10.1101/2021.02.01.429210 ◽

2021 ◽

Author(s):

Nataliya E. Yelina ◽

Sabrina Gonzalez-Jorge ◽

Dominique Hirsz ◽

Ziyi Yang ◽

Ian R. Henderson

Keyword(s):

Meiotic Recombination ◽

Crop Improvement ◽

Double Strand Breaks ◽

Crossover Frequency ◽

Dna Double Strand Breaks ◽

Crop Breeding ◽

Catalytic Subunits ◽

Strand Breaks ◽

Homologous Chromosomes ◽

Meiotic Crossover

AbstractDuring meiosis, homologous chromosomes pair and recombine, which can result in reciprocal crossovers that increase genetic diversity. Crossovers are unevenly distributed along eukaryote chromosomes and show repression in heterochromatin and the centromeres. Within the chromosome arms crossovers are often concentrated in hotspots, which are typically in the kilobase range. The uneven distribution of crossovers along chromosomes, together with their low number per meiosis, creates a limitation during crop breeding, where recombination can be beneficial. Therefore, targeting crossovers to specific genome locations has the potential to accelerate crop improvement. In plants, meiotic crossovers are initiated by DNA double strand breaks (DSBs) that are catalysed by SPO11 complexes, which consist of two catalytic (SPO11-1 and SPO11-2) and two non-catalytic subunits (MTOPVIB). We used the model plant Arabidopsis thaliana to target a dCas9-MTOPVIB fusion protein to the 3a crossover hotspot via CRISPR. We observed that this was insufficient to significantly change meiotic crossover frequency or pattern within 3a. We discuss the implications of our findings for targeting meiotic recombination within plant genomes.

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