Meiotic Knockdown and Complementation Reveals Essential Role of RAD51 in Mouse Spermatogenesis

Dai, Jieqiong; Voloshin, Oleg N.; Potapova, Svetlana; Camerini‐Otero, R. Daniel

doi:10.1016/j.celrep.2017.01.024

Cited by 75 publications

(55 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In humans, researchers have yet to determine why two homologous DNA recombinases, namely, RAD51 and DMC1, are required in mammalian meiosis 40 41. To identify the mechanism of HR, we should create a RAD51 mutant model that specifically disrupts only one of the RAD51-HR-DSB repair functions, which are mitosis and meiosis HR-DSB repair functions.…”

Section: Discussionmentioning

confidence: 99%

XRCC2 mutation causes meiotic arrest, azoospermia and infertility

et al. 2018

View full text Add to dashboard Cite

BackgroundMeiotic homologous recombination (HR) plays an essential role in gametogenesis. In most eukaryotes, meiotic HR is mediated by two recombinase systems: ubiquitous RAD51 and meiosis-specific DMC1. In the RAD51-mediated HR system, RAD51 and five RAD51 paralogues are essential for normal RAD51 function, but the role of RAD51 in human meiosis is unclear. The knockout of Rad51 or any Rad51 paralogue in mice exhibits embryonic lethality. We investigated a family with meiotic arrest, azoospermia and infertility but without other abnormalities.MethodsHomozygosity mapping and whole-exome sequencing were performed in a consanguineous family. An animal model carrying a related mutation was created by using a CRISPR/Cas9 system.ResultsWe identified a 1 bp homozygous substitution (c.41T>C/p.Leu14Pro) on a RAD51 paralogue, namely, XRCC2, in the consanguineous family. We did not detect any XRCC2 recessive mutation in a cohort of 127 males with non-obstructive-azoospermia. Knockin mice with Xrcc2-c.T41C/p.Leu14Pro mutation were generated successfully by the CRISPR/Cas9 method. The homozygotes survived and exhibited meiotic arrest, azoospermia, premature ovarian failure and infertility.ConclusionA XRCC2 recessive mutation causing meiotic arrest and infertility in humans was duplicated with knockin mice. Our results revealed a new Mendelian hereditary entity and provided an experimental model of RAD51-HR gene defect in mammalian meiosis.

show abstract

Section: Discussionmentioning

confidence: 99%

XRCC2 mutation causes meiotic arrest, azoospermia and infertility

et al. 2018

View full text Add to dashboard Cite

show abstract

“…A recent analysis of the Rad51 knockdown in mouse testis showed the depletion of primary spermatocytes and the reduction in crossover formation (Dai et al . ). These facts indicate that both RAD51 and DMC1 are required for meiotic recombination.…”

Section: Introductionmentioning

confidence: 97%

“…In meiosis, RAD51 also acts as an accessory factor to promote DMC1-mediated strand invasion (Cloud et al 2012;Da Ines et al 2013;Kobayashi et al 2014). A recent analysis of the Rad51 knockdown in mouse testis showed the depletion of primary spermatocytes and the reduction in crossover formation (Dai et al 2017). These facts indicate that both RAD51 and DMC1 are required for meiotic recombination.…”

Section: Introductionmentioning

confidence: 99%

SYCP3 regulates strand invasion activities of RAD51 and DMC1

et al. 2017

View full text Add to dashboard Cite

The synaptonemal complex is a higher-ordered proteinaceous architecture formed between homologous chromosomes. SYCP3 is a major component of the lateral/axial elements in the synaptonemal complex and is essential for meiotic recombination. Previous genetic studies showed that SYCP3 functions in meiotic homologous recombination biased to interhomologous chromosomes, by regulating the strand invasion activities of the RAD51 and DMC1 recombinases. However, the mechanism by which SYCP3 regulates RAD51- and DMC1-mediated strand invasion remains elusive. In this study, we found that SYCP3 significantly suppresses the RAD51-mediated, but not the DMC1-mediated, strand invasion reaction by competing with HOP2-MND1, which is an activator for both RAD51 and DMC1. A SYCP3 mutant with defective RAD51 binding does not inhibit the RAD51-mediated homologous recombination in human cells. Therefore, SYCP3 may promote the DMC1-driven homologous recombination by attenuating the RAD51 activity during meiosis.

show abstract

“…A small number of DSBs form crossover recombinations, and the majority of DSBs are repaired as non‐crossover recombinations in spermatocytes 17, 18. These processes involve a number of meiotic‐specific and ubiquitously expressed nuclear proteins, such as γH2AX, RAD51, RAD18, DMC1, BRCA1 and MLH1 19, 20, 21, 22. These proteins are co‐ordinately recruited to DSB sites to facilitate DNA DSB repair via homologous recombination.…”

Section: Introductionmentioning

confidence: 99%

“…17,18 These processes involve a number of meiotic-specific and ubiquitously expressed nuclear proteins, such as cH2AX, RAD51, RAD18, DMC1, BRCA1 and MLH1. [19][20][21][22] These proteins are co-ordinately recruited to DSB sites to facilitate DNA DSB repair via homologous recombination. Defects in DNA DSB repair can cause chromosome mis-segregation underling aneuploidy-related pathologies or meiotic arrest of spermatocytes leading to male infertility.…”

Section: Introductionmentioning

confidence: 99%

Ablation of Ggnbp2 impairs meiotic DNA double‐strand break repair during spermatogenesis in mice

Guo

Liu

et al. 2018

J Cellular Molecular Medi

View full text Add to dashboard Cite

Gametogenetin (GGN) binding protein 2 (GGNBP2) is a zinc finger protein expressed abundantly in spermatocytes and spermatids. We previously discovered that Ggnbp2 resection caused metamorphotic defects during spermatid differentiation and resulted in an absence of mature spermatozoa in mice. However, whether GGNBP2 affects meiotic progression of spermatocytes remains to be established. In this study, flow cytometric analyses showed a decrease in haploid, while an increase in tetraploid spermatogenic cells in both 30‐ and 60‐day‐old Ggnbp2 knockout testes. In spread spermatocyte nuclei, Ggnbp2 loss increased DNA double‐strand breaks (DSB), compromised DSB repair and reduced crossovers. Further investigations demonstrated that GGNBP2 co‐immunoprecipitated with a testis‐enriched protein GGN1. Immunofluorescent staining revealed that both GGNBP2 and GGN1 had the same subcellular localizations in spermatocyte, spermatid and spermatozoa. Ggnbp2 loss suppressed Ggn expression and nuclear accumulation. Furthermore, deletion of either Ggnbp2 or Ggn in GC‐2spd cells inhibited their differentiation into haploid cells in vitro. Overexpression of Ggnbp2 in Ggnbp2 null but not in Ggn null GC‐2spd cells partially rescued the defect coinciding with a restoration of Ggn expression. Together, these data suggest that GGNBP2, likely mediated by its interaction with GGN1, plays a role in DSB repair during meiotic progression of spermatocytes.

show abstract

Meiotic Knockdown and Complementation Reveals Essential Role of RAD51 in Mouse Spermatogenesis

Cited by 75 publications

References 45 publications

XRCC2 mutation causes meiotic arrest, azoospermia and infertility

XRCC2 mutation causes meiotic arrest, azoospermia and infertility

SYCP3 regulates strand invasion activities of RAD51 and DMC1

Ablation of Ggnbp2 impairs meiotic DNA double‐strand break repair during spermatogenesis in mice

Contact Info

Product

Resources

About