From the Graduate School of Environmental and Life Science and.
J Biol Chem. 2013 Nov 1;288(44):31830-41. doi: 10.1074/jbc.M113.492306. Epub 2013 Sep 17.
Skeletal fusions with sterility (sks) is an autosomal recessive mutation of mouse that results in male and female sterility because of defects in gametogenesis. The mutants also have skeletal malformations with fused vertebrae and ribs. We examined testicular phenotypes of sks/sks mice to investigate the defects in spermatogenesis. Histological and immunocytochemical analyses and expression analyses of the marker genes demonstrated that spermatogenesis is arrested at mid to late pachytene stage of meiotic prophase with defective synapsis of the homologous chromosomes. Next, we determined the precise chromosomal localization of the sks locus on a 0.3-Mb region of mouse chromosome 4 by linkage analysis. By sequencing the positional candidate genes in this region and whole exome sequencing, we found a GG to TT nucleotide substitution in exon 6 of the Tmem48 gene that encodes a putative transmembrane protein with six transmembrane domains. The nucleotide substitution causes aberrant splicing, which deletes exon 6 of the Tmem48 transcript. Specific expression of TMEM48 was observed in germ cells of males and females. Furthermore, the phenotypes of the sks mutant were completely rescued by the transgenesis of a genomic fragment containing the wild-type Tmem48 gene. These findings indicate that the Tmem48 mutation is responsible for the gametogenesis defects and skeletal malformations in the sks mice. The TMEM48 protein is a nuclear membrane protein comprising the nuclear pore complex; its exact function in the nuclear pore complex is still unknown. Our finding suggested that the nuclear pore complex plays an important role in mammalian gametogenesis and skeletal development.
Skeletal fusions with sterility (sks) 是一种小鼠的常染色体隐性突变,导致雌雄不育,因为配子发生缺陷。突变体还具有骨骼畸形,椎体和肋骨融合。我们检查了 sks/sks 小鼠的睾丸表型,以研究精子发生缺陷。组织学和免疫细胞化学分析以及标记基因的表达分析表明,精子发生在减数分裂前期的中到晚期粗线期停滞,同源染色体的联会有缺陷。接下来,我们通过连锁分析确定了 sks 基因座在小鼠染色体 4 上 0.3-Mb 区域的精确染色体定位。通过对该区域的位置候选基因进行测序和全外显子组测序,我们发现 Tmem48 基因外显子 6 中的 GG 到 TT 核苷酸取代,该基因编码具有六个跨膜结构域的假定跨膜蛋白。核苷酸取代导致异常剪接,从而删除 Tmem48 转录本的外显子 6。TMEM48 在雄性和雌性生殖细胞中特异性表达。此外,野生型 Tmem48 基因的基因组片段的转基因完全挽救了 sks 突变体的表型。这些发现表明 Tmem48 突变是导致 sks 小鼠配子发生缺陷和骨骼畸形的原因。TMEM48 蛋白是一种核膜蛋白,构成核孔复合物;其在核孔复合物中的确切功能尚不清楚。我们的发现表明核孔复合物在哺乳动物配子发生和骨骼发育中起着重要作用。