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

Zhuang X, Wang L, Song Z, Xiao W - PLoS ONE (2015)

Bottom Line: A significantly reduced solubility was observed for this mutant.We have identified a novel mutation, c.451_452insGACT, in CRYGD, which is associated with nuclear 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Shengjing Hospital, China Medical University, Shenyang, China; Department of Ophthalmology, The Fourth People's Hospital of Shenyang, Shenyang, China.

ABSTRACT

Objective: To investigate a novel insertion variant of CRYGD identified in a Chinese family with nuclear congenital cataract.

Methods: 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.

Principal findings: 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.

Conclusions: 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.

No MeSH data available.


Related in: MedlinePlus

Localization of Myc-tagged wildtype or Y151* CRYGD in HEK293T cells.Immunofluorescence of Myc (green fluorescence) showed the distribution of wildtype CRYGD in both cytomembrane and cytoplasm. However, the mutant CRYGD was mainly localized in the nucleus, in the form of granular deposits. Scale bar: 10 μm.
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pone.0131471.g005: Localization of Myc-tagged wildtype or Y151* CRYGD in HEK293T cells.Immunofluorescence of Myc (green fluorescence) showed the distribution of wildtype CRYGD in both cytomembrane and cytoplasm. However, the mutant CRYGD was mainly localized in the nucleus, in the form of granular deposits. Scale bar: 10 μm.

Mentions: To confirm the pathogenicity of the mutation Y151*, immunofluorescence assay was performed to detect the subcellular localization of mutant CRYGD. Myc-tagged wildtype CRYGD was localized in both cytoplasmic and membrane regions, whereas Y151* was redistributed in the nucleus as a spot-shaped structure (Fig 5). Minimal staining in the cytoplasm was also observed. The aggregation of the mutant protein in the nucleus showed that it failed to form crystallin and perform its intrinsic function.


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

Zhuang X, Wang L, Song Z, Xiao W - PLoS ONE (2015)

Localization of Myc-tagged wildtype or Y151* CRYGD in HEK293T cells.Immunofluorescence of Myc (green fluorescence) showed the distribution of wildtype CRYGD in both cytomembrane and cytoplasm. However, the mutant CRYGD was mainly localized in the nucleus, in the form of granular deposits. Scale bar: 10 μm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493073&req=5

pone.0131471.g005: Localization of Myc-tagged wildtype or Y151* CRYGD in HEK293T cells.Immunofluorescence of Myc (green fluorescence) showed the distribution of wildtype CRYGD in both cytomembrane and cytoplasm. However, the mutant CRYGD was mainly localized in the nucleus, in the form of granular deposits. Scale bar: 10 μm.
Mentions: To confirm the pathogenicity of the mutation Y151*, immunofluorescence assay was performed to detect the subcellular localization of mutant CRYGD. Myc-tagged wildtype CRYGD was localized in both cytoplasmic and membrane regions, whereas Y151* was redistributed in the nucleus as a spot-shaped structure (Fig 5). Minimal staining in the cytoplasm was also observed. The aggregation of the mutant protein in the nucleus showed that it failed to form crystallin and perform its intrinsic function.

Bottom Line: A significantly reduced solubility was observed for this mutant.We have identified a novel mutation, c.451_452insGACT, in CRYGD, which is associated with nuclear 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.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Shengjing Hospital, China Medical University, Shenyang, China; Department of Ophthalmology, The Fourth People's Hospital of Shenyang, Shenyang, China.

ABSTRACT

Objective: To investigate a novel insertion variant of CRYGD identified in a Chinese family with nuclear congenital cataract.

Methods: 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.

Principal findings: 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.

Conclusions: 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.

No MeSH data available.


Related in: MedlinePlus