A Novel Insertion Variant of CRYGD Is Associated with Congenital Nuclear Cataract in a Chinese Family

• Published in: PloS one 2015-07, Vol.10 (7), p.e0131471-e0131471
• Main Authors: Zhuang, Xiaotong, Wang, Lianqing, Song, Zixun, Xiao, Wei
• Format: Article
• Language: English
• Subjects: Asian Continental Ancestry Group - genetics; Cataract - genetics; Cataracts; Cloning; Cytoplasm; Deoxyribonucleic acid; DNA; DNA Mutational Analysis; Female; Frameshift Mutation; gamma-Crystallins - genetics; Gene sequencing; Genes; Genotype; HEK293 Cells; Humans; Identification methods; Immunofluorescence; Insertion; Localization; Mutation; Myc protein; Nuclei; Patients; Pedigree; Polymerase chain reaction; Proteins; Solubility; Western blotting
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0131471

To investigate a novel insertion variant of CRYGD identified in a Chinese family with nuclear congenital cataract. A Chinese family with congenital nuclear cataract was recruited for the mutational screening of candidate genes by direct sequencing. Recombinant N-terminal Myc tagged wildtype or mutant CRYGD was expressed in HEK293T cells. The expression pattern, protein solubility and subcellular distribution were analyzed by western blotting and immunofluorescence. A novel insertion variant, c.451_452insGACT, in CRYGD was identified in the patients. It causes a frameshift and a premature termination of the polypeptide to become Y151*. A significantly reduced solubility was observed for this mutant. Unlike wildtype CRYGD, which existed mainly in the cytoplasm, Y151* was mis-located in the nucleus. We have identified a novel mutation, c.451_452insGACT, in CRYGD, which is associated with nuclear cataract. This is the first insertion mutation of CRYGD found to cause autosomal dominant congenital cataract. The mutant protein, with loss of solubility and localization to the nucleus, is hypothesized to be the major cause of cataract in these patients.