Carroll Renée, Kumar Raman, Shaw Marie, Slee Jennie, Kalscheuer Vera M, Corbett Mark A, Gecz Jozef
The Robinson Research Institute, The University of Adelaide, Adelaide, Australia.
School of Medicine, The University of Adelaide, Adelaide, Australia.
Eur J Hum Genet. 2017 Sep;25(9):1078-1082. doi: 10.1038/ejhg.2017.97. Epub 2017 Jun 14.
Congenital microcephaly, with or without additional developmental defects, is a heterogeneous disorder resulting from impaired brain development during early fetal life. The majority of causative genetic variants identified thus far are inherited in an autosomal recessive manner and impact key cellular pathways such as mitosis, DNA damage response and repair, apoptosis and splicing. Here, we report a novel donor splice site variant in the G-patch domain and KOW motifs (GPKOW) gene (NG_021310.2:g.6126G>A, NM_015698.4:c.331+5G>A) that segregates with affected and carrier status in a multigenerational family with an X-linked perinatal lethal condition characterized by severe microcephaly and intrauterine growth restriction (IUGR). GPKOW is a core member of the spliceosome that has been shown in numerous model organisms and in human cells to be essential for survival. By investigating GPKOW transcripts in lymphoblastoid cell lines (LCLs) of three carrier females, we show that the GPKOW c.331+5G>A variant disrupts normal splicing of its pre-mRNAs. In a clonal culture expressing only the c.331+5G>A allele isolated from one carrier female LCL, we observed an 80% reduction in wild type GPKOW mRNA, 70% reduction in the full length GPKOW protein and the presence of a truncated GPKOW protein with possible dominant negative effect. Based on our and published data we propose that the GPKOW gene is essential for fetal development and when disrupted, leads to a severe, male-lethal phenotype characterised by microcephaly and IUGR.
先天性小头畸形,伴或不伴有其他发育缺陷,是一种异质性疾病,由胎儿早期大脑发育受损引起。迄今为止确定的大多数致病基因变异以常染色体隐性方式遗传,并影响关键的细胞途径,如有丝分裂、DNA损伤反应与修复、细胞凋亡和剪接。在此,我们报告了G-补丁结构域和KOW模体(GPKOW)基因中的一个新的供体剪接位点变异(NG_021310.2:g.6126G>A,NM_015698.4:c.331+5G>A),该变异在一个多代家族中与受影响状态和携带者状态分离,该家族患有X连锁围产期致死性疾病,其特征为严重小头畸形和宫内生长受限(IUGR)。GPKOW是剪接体的核心成员,在众多模式生物和人类细胞中已表明其对生存至关重要。通过研究三名携带者女性淋巴母细胞系(LCL)中的GPKOW转录本,我们发现GPKOW c.331+5G>A变异破坏了其前体mRNA的正常剪接。在仅表达从一名携带者女性LCL分离出的c.331+5G>A等位基因的克隆培养物中,我们观察到野生型GPKOW mRNA减少了80%,全长GPKOW蛋白减少了70%,并且存在一种可能具有显性负效应的截短GPKOW蛋白。基于我们的研究和已发表的数据,我们提出GPKOW基因对胎儿发育至关重要,当其被破坏时,会导致一种以小头畸形和IUGR为特征的严重男性致死性表型。