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1.
Nucleic Acids Res ; 51(16): 8606-8622, 2023 09 08.
Article in English | MEDLINE | ID: mdl-37439366

ABSTRACT

Recruitment of RAD51 and/or DMC1 recombinases to single-strand DNA is indispensable for homology search and strand invasion in homologous recombination (HR) and for protection of nascent DNA strands at stalled replication forks. Thereafter RAD51/DMC1 dissociate, actively or passively, from these joint molecules upon DNA repair or releasing from replication stress. However, the mechanism that regulates RAD51/DMC1 dissociation and its physiological importance remain elusive. Here, we show that a FLIP-FIGNL1 complex regulates RAD51 and DMC1 dissociation to promote meiotic recombination and replication fork restart in mammals. Mice lacking FLIP are embryonic lethal, while germline-specific deletion of FLIP leads to infertility in both males and females. FLIP-null meiocytes are arrested at a zygotene-like stage with massive RAD51 and DMC1 foci, which frequently co-localize with SHOC1 and TEX11. Furthermore, FLIP interacts with FIGNL1. Depletion of FLIP or FIGNL1 in cell lines destabilizes each other and impairs RAD51 dissociation. Thus, the active dissociation of RAD51/DMC1 by the FLIP-FIGNL1 complex is a crucial step required for HR and replication fork restart, and represents a conserved mechanism in somatic cells and germ cells.


Subject(s)
DNA-Binding Proteins , Rad51 Recombinase , Male , Female , Animals , Mice , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Rad51 Recombinase/genetics , Rad51 Recombinase/metabolism , Homologous Recombination/genetics , DNA Replication , DNA/metabolism , Cell Cycle Proteins/genetics , Cell Cycle Proteins/metabolism , Meiosis/genetics , Mammals/genetics
2.
Cell Mol Life Sci ; 80(6): 152, 2023 May 17.
Article in English | MEDLINE | ID: mdl-37198331

ABSTRACT

The sperm flagellum is a specialized type of motile cilium composed of a typical "9 + 2" axonemal structure with peri-axonemal structures, such as outer dense fibers (ODFs). This flagellar arrangement is crucial for sperm movement and fertilization. However, the association of axonemal integrity with ODFs remains poorly understood. Here, we demonstrate that mouse BBOF1 could interact with both MNS1, an axonemal component, and ODF2, an ODF protein, and is required for sperm flagellar axoneme maintenance and male fertility. BBOF1 is expressed exclusively in male germ cells from the pachytene stage onwards and is detected in sperm axoneme fraction. Spermatozoa derived from Bbof1-knockout mice exhibit a normal morphology, however, reduced motility due to the absence of certain microtubule doublets, resulting in the failure to fertilize mature oocytes. Furthermore, BBOF1 is found to interact with ODF2 and MNS1 and is also required for their stability. Our findings in mice suggest that Bbof1 could also be essential for human sperm motility and male fertility, thus is a novel potential candidate gene for asthenozoospermia diagnosis.


Subject(s)
Axoneme , Infertility, Male , Animals , Male , Mice , Axoneme/metabolism , Fertility/genetics , Heat-Shock Proteins/metabolism , Infertility, Male/genetics , Infertility, Male/metabolism , Mice, Knockout , Semen/metabolism , Sperm Motility/genetics , Spermatozoa/metabolism
3.
BMC Plant Biol ; 21(1): 144, 2021 Mar 19.
Article in English | MEDLINE | ID: mdl-33740889

ABSTRACT

BACKGROUND: Identifying the loci and dissecting the genetic architecture underlying wheat yield- and quality-related traits are essential for wheat breeding. A genome-wide association study was conducted using a high-density 90 K SNP array to analyze the yield- and quality-related traits of 543 bread wheat varieties. RESULTS: A total of 11,140 polymorphic SNPs were distributed on 21 chromosomes, including 270 significant SNPs associated with 25 yield- and quality-related traits. Additionally, 638 putative candidate genes were detected near the significant SNPs based on BLUP data, including three (TraesCS7A01G482000, TraesCS4B01G343700, and TraesCS6B01G295400) related to spikelet number per spike, diameter of the first internode, and grain volume. The three candidate genes were further analyzed using stage- and tissue- specific gene expression data derived from an RNA-seq analysis. These genes are promising candidates for enhancing yield- and quality-related traits in wheat. CONCLUSIONS: The results of this study provide a new insight to understand the genetic basis of wheat yield and quality. Furthermore, the markers detected in this study may be applicable for marker-assisted selection in wheat breeding programs.


Subject(s)
Triticum/growth & development , Triticum/genetics , Edible Grain/genetics , Edible Grain/growth & development , Genetic Markers , Genome-Wide Association Study , Phenotype , Polymorphism, Single Nucleotide
4.
Int J Mol Sci ; 20(16)2019 Aug 12.
Article in English | MEDLINE | ID: mdl-31409051

ABSTRACT

It is now widely accepted that allele-specific DNA methylation (ASM) commonly occurs at non-imprinted loci. Most of the non-imprinted ASM regions observed both within and outside of the CpG island show a strong correlation with DNA polymorphisms. However, what polymorphic cis-acting elements mediate non-imprinted ASM of the CpG island remains unclear. In this study, we investigated the impact of polymorphic GT microsatellites within the gene promoter on non-imprinted ASM of the local CpG island in goldfish. We generated various goldfish heterozygotes, in which the length of GT microsatellites or some non-repetitive sequences in the promoter of no tail alleles was different. By examining the methylation status of the downstream CpG island in these heterozygotes, we found that polymorphisms of a long GT microsatellite can lead to the ASM of the downstream CpG island during oogenesis and embryogenesis, polymorphisms of short GT microsatellites and non-repetitive sequences in the promoter exhibited no significant effect on the methylation of the CpG island. We also observed that the ASM of the CpG island was associated with allele-specific expression in heterozygous embryos. These results suggest that a long polymorphic GT microsatellite within a gene promoter mediates non-imprinted ASM of the local CpG island in a goldfish inter-strain hybrid.


Subject(s)
DNA Methylation , Goldfish/genetics , Microsatellite Repeats , Alleles , Animals , Chimera/genetics , CpG Islands , Crosses, Genetic , Female , Genomic Imprinting , Goldfish/embryology , Male , Polymorphism, Genetic , Promoter Regions, Genetic
5.
Nanoscale ; 9(38): 14357-14363, 2017 Oct 05.
Article in English | MEDLINE | ID: mdl-28726939

ABSTRACT

In this paper, we fabricate a graphene film heater through laser reduction on graphene oxide, which is a two-step process. The electrothermal performance of the graphene heater can be adjusted by the laser energy density. While the applied voltage is 18 V, the graphene heater reaches a steady-state temperature of 247.3 °C within 20 s. After the graphene heater is folded in half 100 times, its output temperature remains to be precisely controlled by the input power and the temperature distribution is uniform. In addition, the flexibility of the graphene heater is superior to a heater based on a commercial indium tin oxide film. It's worth noting that the graphene heater can be fabricated with desired shapes directly and easily, which is rare among the reported film heaters. In consideration of the high performance of the graphene film heater, we demonstrate its three application scenarios: portable warmers applied in medical infusion apparatus, flexible custom-shaped heaters for special requirements and displays.

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