Structural and functional analyses of the DMC1-M200V polymorphism found in the human population

Abstract

The M200V polymorphism of the human DMC1 protein, which is an essential, meiosis-specific DNA recombinase, was found in an infertile patient, raising the question of whether this homozygous human DMC1-M200V polymorphism may cause infertility by affecting the function of the human DMC1 protein. In the present study, we determined the crystal structure of the human DMC1-M200V variant in the octameric-ring form. Biochemical analyses revealed that the human DMC1-M200V variant had reduced stability, and was moderately defective in catalyzing in vitro recombination reactions. The corresponding M194V mutation introduced in the Schizosaccharomyces pombe dmc1 gene caused a significant decrease in the meiotic homologous recombination frequency. Together, these structural, biochemical and genetic results provide extensive evidence that the human DMC1-M200V mutation impairs its function, supporting the previous interpretation that this single-nucleotide polymorphism is a source of human infertility.

DMC1 mutation that causes human non-obstructive azoospermia and premature ovarian insufficiency identified by whole-exome sequencing

“…In vitro structural and functional analyses showed that DMC1 -M200V was a disease-causing mutation 34. However, the pathogenicity of the variant has been later questioned owing to its high frequency in the African population, and has been found as homozygous 172 times in African and Latino populations according to the gnomAD database.…”

Green fluorescent protein fused to the C terminus of RAD51 specifically interferes with secondary DNA binding by the RAD51-ssDNA complex

“…Purification of proteins Rice (Oryza sativa) RAD51A1, rice RAD51A2, human RAD51, rice DMC1A and human DMC1 were prepared by methods described previously (Matsuo et al, 2006;Hikiba et al, 2008;Ishida et al, 2008;Sakane et al, 2008;Morozumi et al, 2013). For the RAD51-GFP proteins, the stop codons of rice RAD51A1, rice RAD51A2 and human RAD51 genes were replaced by a DNA fragment encoding Pro-Val-Ala-Thr as a linker peptide, and the enhanced GFP (EGFP) gene was fused just after the linker peptide sequence.…”

Molecular characterization and epigenetic regulation of Mei1 in cattle and cattle–yak

“…In human, DMC1 variants (e.g. DMC1-M200V) were the source of infertility (Hikiba et al, 2008). However, little was known about the relationship between Mei and male infertility.…”