A meiosis-specific factor C19orf57/4930432K21Rik/BRME1 modulates localization of RAD51 and DMC1 recombinases to DSBs in mouse meiotic recombination

Takemoto, Kazumasa; Tani, Naoki; Takada-Horisawa, Yuki; Fujimura, Shigefumi; Tanno, Nobuhiro; Yamane, Mariko; Okamura, Kaho; Sugimoto, Michihiko; Araki, Kimi; Ishiguro, Kei‐ichiro

doi:10.1101/2020.02.14.950204

Cited by 4 publications

(6 citation statements)

References 47 publications

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“…In the mutant, RPA foci were present and the number was increased compared to wild type (Figure 2D,E), suggesting that DSBs can be formed, but that RPA cannot be replaced by recombinases. Also, the foci numbers of SPATA22, a meiosis-specific single-stranded DNA binding protein [34][35][36] , were higher in absence of exons 12-14 of Brca2 than in the controls (Figure 2D,F), similar to what was observed in Hsf2bp -/and Brme1 -/spermatocytes 19,25,27,28 . Since the Brca2 ∆12-14 mouse model completely lacks the HSF2BP-binding domain, we investigated the localization patterns of HSF2BP and its interaction partner BRME1.…”

Section: Introductionsupporting

confidence: 77%

“…Mice deficient for HSF2BP are born at Mendelian ratios and have no overt somatic phenotypes, but males are infertile due to meiotic HR failure. Shortly after, HSF2BP was shown to directly interact with another uncharacterized protein named BRME1 (also called MAMERR or MEIOK21), and the phenotypes of the Hsf2bp and Brme1 knockout mouse models 19,21,[25][26][27][28][29] were similar.…”

Section: Introductionmentioning

confidence: 99%

“…Hsf2bp -/and Brme1 -/mice showed a sexual dimorphism regarding defects in meiosis 19,21,[25][26][27][28][29]37 . Therefore, we also analyzed meiotic HR protein localization patterns in embryonic (E15.5 and E17.5) Brca2 ∆12-14 oocyte nuclei, since female meiotic prophase initiates during embryogenesis.…”

Section: Introductionmentioning

confidence: 99%

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Chromosome Pairing Through Tensioned DNA Tethers Revealed by BRCA2 Meiotic Domain Deletion

Koornneef,

van Rossum-Fikkert,

Sleddens-Linkels

et al. 2023

Preprint

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BRCA2 has multiple functional domains that interact with different partners, and is essential for both somatic and meiotic homologous recombination (HR). We created aBrca2Δ12-14mouse model with an internal deletion of the region which we named “the meiotic domain of BRCA2”, as its loss results in complete failure of meiotic HR, while somatic HR is intact. The deletion in the protein includes the HSF2BP-binding motifs (exons 12-13) and the DMC1-binding PhePP domain (exon 14).Brca2Δ12-14mice showed complete infertility in both males and females, with sexually dimorphic features. Recombinase foci (both RAD51 and DMC1) were completely undetectable in mutant spermatocytes, but while DMC1 foci were also absent in mutant oocytes, RAD51 foci numbers were only partially reduced (up to 60% fewer, 20% less intense). The relevance of the PhePP domain for meiotic HR has been controversial, but both the phenotype ofBrca2Δ12-14, and our biochemical data indicate that, along with the BRC repeats of BRCA2, PhePP is both essential and specific for DMC1 loading in meiotic HR, analogous to the C-terminal RAD51-specific TR2/CTRB. Further investigation of DSB end processing inBrca2Δ12-14meiocytes and controls, using super-resolution imaging of RPA and SYCP3 led to discovery of two novel features. First, inBrca2Δ12-14oocytes, but not in the spermatocytes nor wild types, we observed RPA foci as doublets ∼200 nm apart, which could represent DSB end separation. Second, we describe RPA structures that are completely HR-dependent and are indicative of long, double-stranded DNA connections between homologs prior to synapsis. The observations led us to a model for chromosome alignment via multiple, tensed DNA tethers that connect the homologous DNA regions. We propose that in combination with dynamic adjustment of chromatin loops by meiotic cohesins, this is a plausible molecular mechanism to bring the homologs closer and initiate synapsis.

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Section: Introductionsupporting

confidence: 77%

Section: Introductionmentioning

confidence: 99%

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Chromosome Pairing Through Tensioned DNA Tethers Revealed by BRCA2 Meiotic Domain Deletion

Koornneef,

van Rossum-Fikkert,

Sleddens-Linkels

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…Surface-spread nuclei from spermatocytes were prepared by the dry down method as described (Peters et al, 1997) (Takemoto et al, 2020) with modification. The slides were then air-dried and washed with water containing 0.1 % TritonX100 or frozen for longer storage at -30ºC.…”

Section: Immunostaining Of Spermatocytesmentioning

confidence: 99%

FBXO47 is essential for preventing the synaptonemal complex from premature disassembly in mouse male meiosis

Ishiguro

Tanno

Takemoto

et al. 2021

Preprint

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Meiotic prophase is a prolonged G2 phase that ensures the completion of numerous meiosis-specific chromosome events. During meiotic prophase, homologous chromosomes undergo synapsis to facilitate meiotic recombination yielding crossovers. It remains largely elusive how homolog synapsis is temporally maintained and destabilized during meiotic prophase. Here we show that FBXO47 is the stabilizer of synaptonemal complex during male meiotic prophase. Disruption of FBXO47 shows severe impact on homologous chromosome synapsis and DSB repair processes, leading to male infertility. Notably, in the absence of FBXO47, although once homologous chromosomes are synapsed, the synaptonemal complex is precociously disassembled before progressing beyond pachytene. Remarkably, Fbxo47 KO spermatocytes remain in earlier stage of meiotic prophase and lack crossovers, despite apparently exhibiting diplotene-like chromosome morphology. We propose that FBXO47 functions independently of SCF E3 ligase, and plays a crucial role in preventing synaptonemal complex from premature disassembly during cell cycle progression of meiotic prophase.

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“…Indeed, SWSAP1-SWS1-SPIDR is necessary for the efficient formation of RAD51/DMC1 focus, which is a cytologically detectable immuno-stained structure, in male meiosis 18,53,54 . Moreover, a meiosis-specific HSF2BP(MEILB2)-BRME1(MEIOK21) complex promotes the assembly of RAD51/DMC1 working together with BRCA2 mainly in male meiotic recombination [55][56][57][58] . Like RAD51 mediators, KO mice of some negative regulators such as BLM are also embryonically lethal 59 while Rad54 KO mice are viable and fertile 60 .…”

Section: Introductionmentioning

confidence: 99%

FIGNL1 AAA+ ATPase remodels RAD51 and DMC1 filaments in pre-meiotic DNA replication and meiotic recombination

Ito

Furukohri

Matsuzaki

et al. 2023

Preprint

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The formation of RAD51/DMC1 filaments on single-stranded (ss)DNAs, which is essential for homology search and strand exchange in DNA double-strand break (DSB) repair and for the protection of stalled DNA replication forks, is tightly regulated in time and space. FIGNL1 AAA+++ ATPase plays positive and negative roles in the RAD51-mediated recombination in human cells. However, the role of FIGNL1 in gametogenesis remains unsolved. Here, we characterized a germ-line-specific conditional knockout (cKO) mouse of FIGNL1. The Fignl1 cKO male mice showed defective chromosome synapsis and impaired meiotic DSB repair with the accumulation of RAD51/DMC1 on chromosomes in mid-meiotic prophase I, supporting a role of FIGNL1 in a post-assembly stage of RAD51/DMC1 filaments. Fignl1 cKO spermatocytes accumulate RAD51 and DMC1 ensembles on chromosomes not only in early meiotic prophase I but also in meiotic S-phase. These RAD51/DMC1 assemblies are independent of meiotic DSB formation. Finally, we showed that purified FIGNL1 dismantles RAD51 filament on double-stranded (ds)DNA as well as ssDNA. These results suggest a critical role of FIGNL1 to limit the uncontrolled assembly of RAD51 and DMC1 on native dsDNAs during the meiotic S-phase and meiotic prophase I.

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A meiosis-specific factor C19orf57/4930432K21Rik/BRME1 modulates localization of RAD51 and DMC1 recombinases to DSBs in mouse meiotic recombination

Cited by 4 publications

References 47 publications

Chromosome Pairing Through Tensioned DNA Tethers Revealed by BRCA2 Meiotic Domain Deletion

Chromosome Pairing Through Tensioned DNA Tethers Revealed by BRCA2 Meiotic Domain Deletion

FBXO47 is essential for preventing the synaptonemal complex from premature disassembly in mouse male meiosis

FIGNL1 AAA+ ATPase remodels RAD51 and DMC1 filaments in pre-meiotic DNA replication and meiotic recombination

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