Chen Weirong, Chen Xiaoyun, Hu Zhengmao, Lin Haotian, Zhou Fengqi, Luo Lixia, Zhang Xinyu, Zhong Xiaojian, Yang Ye, Wu Changrui, Lin Zhuoling, Ye Shaobi, Liu Yizhi
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, People's Republic of China.
PLoS One. 2013 Nov 28;8(11):e81290. doi: 10.1371/journal.pone.0081290. eCollection 2013.
Congenital cataract is a major cause of visual impairment and childhood blindness. The solubility and stability of crystallin proteins play critical roles in maintaining the optical transparency of the lens during the life span. Previous studies have shown that approximately 8.3%~25% of congenital cataracts are inherited, and mutations in crystallins are the most common. In this study, we attempted to identify the genetic defect in a four-generation family affected with congenital cataracts. The congenital cataract phenotype of this four-generation family was identified as membranous cataract by slit-lamp photography. Mutation screening of the candidate genes detected a heterozygous c.465G → C change in the exon6 of the βB2-crystallin gene (CRYBB2) in all family members affected with cataracts, resulting in the substitution of a highly conserved Tryptophan to Cystine (p.W151C). The mutation was confirmed by restriction fragment length polymorphism (RFLP) analysis and found that the transition resulted in the absence of a BslI restriction site in the affected members of the pedigree. The outcome of PolyPhen-2 and SIFT analysis predicted that this W151C mutation would probably damage to the structure and function of βB2-crystallin. Wild type (wt) and W151C mutant βB2-crystallin were expressed in human lens epithelial cells (HLECs), and the fluorescence results showed that Wt-βB2-crystallin was evenly distributed throughout the cells, whereas approximately 34.7% of cells transfected with the W151C mutant βB2-crystallin formed intracellular aggregates. Taken together, these data suggest that the missense mutation in CRYBB2 gene leads to progressive congenital membranous cataract by impacting the solubility and function of βB2-crystallin.
先天性白内障是视力损害和儿童失明的主要原因。晶状体蛋白的溶解性和稳定性在维持晶状体一生的光学透明度方面起着关键作用。先前的研究表明,约8.3%~25%的先天性白内障是遗传性的,其中晶状体蛋白的突变最为常见。在本研究中,我们试图确定一个患先天性白内障的四代家系中的基因缺陷。通过裂隙灯照相术,将这个四代家系的先天性白内障表型确定为膜性白内障。对候选基因进行突变筛查,在所有患白内障的家庭成员中检测到βB2-晶状体蛋白基因(CRYBB2)外显子6上存在杂合的c.465G→C变化,导致一个高度保守的色氨酸被胱氨酸取代(p.W151C)。通过限制性片段长度多态性(RFLP)分析证实了该突变,发现在家系中受影响的成员中这种转变导致BslI限制性位点缺失。PolyPhen-2和SIFT分析结果预测,这种W151C突变可能会损害βB2-晶状体蛋白的结构和功能。野生型(wt)和W151C突变型βB2-晶状体蛋白在人晶状体上皮细胞(HLECs)中表达,荧光结果显示,wt-βB2-晶状体蛋白均匀分布于整个细胞中,而转染W151C突变型βB2-晶状体蛋白的细胞中约34.7%形成细胞内聚集体。综上所述,这些数据表明CRYBB2基因中的错义突变通过影响βB2-晶状体蛋白的溶解性和功能导致进行性先天性膜性白内障。