A novel human CRYGD mutation in a juvenile autosomal dominant cataract

Mascarenhas Roshan, Pai H. Vijaya, G. Rao Lavanya, Prasada K. Shama, S. T. Santhiya, Jochen Graw, P. M. Gopinath, K. Satyamoorthy

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Purpose: Identification of causal mutation in the crystallin, connexin, and paired box 6 (PAX6) genes associated with childhood cataract in patients from India. Methods: In this study, forty eight members from seventeen families and 148 sporadic cases of childhood cataract were evaluated. Clinical and ophthalmologic examinations were performed on available affected and unaffected family members. Samples of genomic DNA were PCR amplified to screen for mutations in the candidate genes viz., alpha-A crystallin (CRYAA), beta-B2 crystallin (CRYBB2), gamma-A crystallin (CRYGA), gamma-B crystallin (CRYGB), gamma-C crystallin (CRYGC), gamma-D crystallin (CRYGD), gap junction alpha-3 (GJA3), gap junction alpha-8 (GJA8), and PAX6 based on polymerase chain reaction and single strand conformation polymorphism (PCR-SSCP) analysis. Samples showing any band mobility shift were subjected to bidirectional sequencing to confirm the variation. Co-segregation of the observed change with the disease phenotype was further tested by restriction fragment length polymorphism (RFLP) for the appropriate restriction site. Results: DNA sequencing analysis of CRYAA, CRYBB2, CRYGA-D, GJA3, GJA8, and PAX6 of the affected members of a family (C-35) showed a novel heterozygous missense mutation C>A at position 229 in CRYGD in three affected members of family C-35 with anterior polar coronary cataract. This variation C229A substitution created a novel restriction site for AluI and resulted in a substitution of highly conserved arginine at position 77 by serine (R77S). AluI restriction site analysis confirmed the transversion mutation. Analysis of the available unaffected members of the family (C-35) and 100 unrelated control subjects (200 chromosomes) of the same ethnic background did not show R77S variation. Data generated using ProtScale and PyMOL programs revealed that the mutation altered the stability and solvent-accessibility of the CRYGD protein. Conclusions: We describe here a family having anterior polar coronary cataract that co-segregates with the novel allele R77S of CRYGD in all the affected members. The same was found to be absent in the ethnically matched controls (n=100) studied. Interestingly the residue Arg has been frequently implicated in four missense (R15C, R15S, R37S, and R59H) and in one truncation mutation (R140X) of CRYGD. In two of the reported mutations Arg residues have been replaced with Serine. This finding further expands the mutation spectrum of CRYGD in association with childhood cataract and demonstrates a possible mechanism of cataractogenesis. Screening of other familial (n=48) and sporadic (n=148) cases of childhood cataract, did not reveal any previously reported or novel mutation in the candidate genes screened.

Original languageEnglish
Pages (from-to)887-896
Number of pages10
JournalMolecular Vision
Volume16
Publication statusPublished - 2010

Fingerprint

gamma-Crystallins
Mutation
Gap Junctions
Cataract
Serine
alpha-Crystallin A Chain
human CRYGD protein
Autosomal Dominant Cataract
Genes
Polymerase Chain Reaction
Crystallins
Connexins
Missense Mutation
DNA Sequence Analysis
Restriction Fragment Length Polymorphisms
Arginine
India
Chromosomes
Alleles

All Science Journal Classification (ASJC) codes

  • Ophthalmology

Cite this

Roshan, M., Vijaya, P. H., Lavanya, G. R., Shama, P. K., Santhiya, S. T., Graw, J., ... Satyamoorthy, K. (2010). A novel human CRYGD mutation in a juvenile autosomal dominant cataract. Molecular Vision, 16, 887-896.
Roshan, Mascarenhas ; Vijaya, Pai H. ; Lavanya, G. Rao ; Shama, Prasada K. ; Santhiya, S. T. ; Graw, Jochen ; Gopinath, P. M. ; Satyamoorthy, K. / A novel human CRYGD mutation in a juvenile autosomal dominant cataract. In: Molecular Vision. 2010 ; Vol. 16. pp. 887-896.
@article{6d251b22d05047b29b80a16cf1cebcbb,
title = "A novel human CRYGD mutation in a juvenile autosomal dominant cataract",
abstract = "Purpose: Identification of causal mutation in the crystallin, connexin, and paired box 6 (PAX6) genes associated with childhood cataract in patients from India. Methods: In this study, forty eight members from seventeen families and 148 sporadic cases of childhood cataract were evaluated. Clinical and ophthalmologic examinations were performed on available affected and unaffected family members. Samples of genomic DNA were PCR amplified to screen for mutations in the candidate genes viz., alpha-A crystallin (CRYAA), beta-B2 crystallin (CRYBB2), gamma-A crystallin (CRYGA), gamma-B crystallin (CRYGB), gamma-C crystallin (CRYGC), gamma-D crystallin (CRYGD), gap junction alpha-3 (GJA3), gap junction alpha-8 (GJA8), and PAX6 based on polymerase chain reaction and single strand conformation polymorphism (PCR-SSCP) analysis. Samples showing any band mobility shift were subjected to bidirectional sequencing to confirm the variation. Co-segregation of the observed change with the disease phenotype was further tested by restriction fragment length polymorphism (RFLP) for the appropriate restriction site. Results: DNA sequencing analysis of CRYAA, CRYBB2, CRYGA-D, GJA3, GJA8, and PAX6 of the affected members of a family (C-35) showed a novel heterozygous missense mutation C>A at position 229 in CRYGD in three affected members of family C-35 with anterior polar coronary cataract. This variation C229A substitution created a novel restriction site for AluI and resulted in a substitution of highly conserved arginine at position 77 by serine (R77S). AluI restriction site analysis confirmed the transversion mutation. Analysis of the available unaffected members of the family (C-35) and 100 unrelated control subjects (200 chromosomes) of the same ethnic background did not show R77S variation. Data generated using ProtScale and PyMOL programs revealed that the mutation altered the stability and solvent-accessibility of the CRYGD protein. Conclusions: We describe here a family having anterior polar coronary cataract that co-segregates with the novel allele R77S of CRYGD in all the affected members. The same was found to be absent in the ethnically matched controls (n=100) studied. Interestingly the residue Arg has been frequently implicated in four missense (R15C, R15S, R37S, and R59H) and in one truncation mutation (R140X) of CRYGD. In two of the reported mutations Arg residues have been replaced with Serine. This finding further expands the mutation spectrum of CRYGD in association with childhood cataract and demonstrates a possible mechanism of cataractogenesis. Screening of other familial (n=48) and sporadic (n=148) cases of childhood cataract, did not reveal any previously reported or novel mutation in the candidate genes screened.",
author = "Mascarenhas Roshan and Vijaya, {Pai H.} and Lavanya, {G. Rao} and Shama, {Prasada K.} and Santhiya, {S. T.} and Jochen Graw and Gopinath, {P. M.} and K. Satyamoorthy",
year = "2010",
language = "English",
volume = "16",
pages = "887--896",
journal = "Molecular Vision",
issn = "1090-0535",

}

Roshan, M, Vijaya, PH, Lavanya, GR, Shama, PK, Santhiya, ST, Graw, J, Gopinath, PM & Satyamoorthy, K 2010, 'A novel human CRYGD mutation in a juvenile autosomal dominant cataract' Molecular Vision, vol. 16, pp. 887-896.

A novel human CRYGD mutation in a juvenile autosomal dominant cataract. / Roshan, Mascarenhas; Vijaya, Pai H.; Lavanya, G. Rao; Shama, Prasada K.; Santhiya, S. T.; Graw, Jochen; Gopinath, P. M.; Satyamoorthy, K.

In: Molecular Vision, Vol. 16, 2010, p. 887-896.

Research output: Contribution to journalArticle

TY - JOUR

T1 - A novel human CRYGD mutation in a juvenile autosomal dominant cataract

AU - Roshan, Mascarenhas

AU - Vijaya, Pai H.

AU - Lavanya, G. Rao

AU - Shama, Prasada K.

AU - Santhiya, S. T.

AU - Graw, Jochen

AU - Gopinath, P. M.

AU - Satyamoorthy, K.

PY - 2010

Y1 - 2010

N2 - Purpose: Identification of causal mutation in the crystallin, connexin, and paired box 6 (PAX6) genes associated with childhood cataract in patients from India. Methods: In this study, forty eight members from seventeen families and 148 sporadic cases of childhood cataract were evaluated. Clinical and ophthalmologic examinations were performed on available affected and unaffected family members. Samples of genomic DNA were PCR amplified to screen for mutations in the candidate genes viz., alpha-A crystallin (CRYAA), beta-B2 crystallin (CRYBB2), gamma-A crystallin (CRYGA), gamma-B crystallin (CRYGB), gamma-C crystallin (CRYGC), gamma-D crystallin (CRYGD), gap junction alpha-3 (GJA3), gap junction alpha-8 (GJA8), and PAX6 based on polymerase chain reaction and single strand conformation polymorphism (PCR-SSCP) analysis. Samples showing any band mobility shift were subjected to bidirectional sequencing to confirm the variation. Co-segregation of the observed change with the disease phenotype was further tested by restriction fragment length polymorphism (RFLP) for the appropriate restriction site. Results: DNA sequencing analysis of CRYAA, CRYBB2, CRYGA-D, GJA3, GJA8, and PAX6 of the affected members of a family (C-35) showed a novel heterozygous missense mutation C>A at position 229 in CRYGD in three affected members of family C-35 with anterior polar coronary cataract. This variation C229A substitution created a novel restriction site for AluI and resulted in a substitution of highly conserved arginine at position 77 by serine (R77S). AluI restriction site analysis confirmed the transversion mutation. Analysis of the available unaffected members of the family (C-35) and 100 unrelated control subjects (200 chromosomes) of the same ethnic background did not show R77S variation. Data generated using ProtScale and PyMOL programs revealed that the mutation altered the stability and solvent-accessibility of the CRYGD protein. Conclusions: We describe here a family having anterior polar coronary cataract that co-segregates with the novel allele R77S of CRYGD in all the affected members. The same was found to be absent in the ethnically matched controls (n=100) studied. Interestingly the residue Arg has been frequently implicated in four missense (R15C, R15S, R37S, and R59H) and in one truncation mutation (R140X) of CRYGD. In two of the reported mutations Arg residues have been replaced with Serine. This finding further expands the mutation spectrum of CRYGD in association with childhood cataract and demonstrates a possible mechanism of cataractogenesis. Screening of other familial (n=48) and sporadic (n=148) cases of childhood cataract, did not reveal any previously reported or novel mutation in the candidate genes screened.

AB - Purpose: Identification of causal mutation in the crystallin, connexin, and paired box 6 (PAX6) genes associated with childhood cataract in patients from India. Methods: In this study, forty eight members from seventeen families and 148 sporadic cases of childhood cataract were evaluated. Clinical and ophthalmologic examinations were performed on available affected and unaffected family members. Samples of genomic DNA were PCR amplified to screen for mutations in the candidate genes viz., alpha-A crystallin (CRYAA), beta-B2 crystallin (CRYBB2), gamma-A crystallin (CRYGA), gamma-B crystallin (CRYGB), gamma-C crystallin (CRYGC), gamma-D crystallin (CRYGD), gap junction alpha-3 (GJA3), gap junction alpha-8 (GJA8), and PAX6 based on polymerase chain reaction and single strand conformation polymorphism (PCR-SSCP) analysis. Samples showing any band mobility shift were subjected to bidirectional sequencing to confirm the variation. Co-segregation of the observed change with the disease phenotype was further tested by restriction fragment length polymorphism (RFLP) for the appropriate restriction site. Results: DNA sequencing analysis of CRYAA, CRYBB2, CRYGA-D, GJA3, GJA8, and PAX6 of the affected members of a family (C-35) showed a novel heterozygous missense mutation C>A at position 229 in CRYGD in three affected members of family C-35 with anterior polar coronary cataract. This variation C229A substitution created a novel restriction site for AluI and resulted in a substitution of highly conserved arginine at position 77 by serine (R77S). AluI restriction site analysis confirmed the transversion mutation. Analysis of the available unaffected members of the family (C-35) and 100 unrelated control subjects (200 chromosomes) of the same ethnic background did not show R77S variation. Data generated using ProtScale and PyMOL programs revealed that the mutation altered the stability and solvent-accessibility of the CRYGD protein. Conclusions: We describe here a family having anterior polar coronary cataract that co-segregates with the novel allele R77S of CRYGD in all the affected members. The same was found to be absent in the ethnically matched controls (n=100) studied. Interestingly the residue Arg has been frequently implicated in four missense (R15C, R15S, R37S, and R59H) and in one truncation mutation (R140X) of CRYGD. In two of the reported mutations Arg residues have been replaced with Serine. This finding further expands the mutation spectrum of CRYGD in association with childhood cataract and demonstrates a possible mechanism of cataractogenesis. Screening of other familial (n=48) and sporadic (n=148) cases of childhood cataract, did not reveal any previously reported or novel mutation in the candidate genes screened.

UR - http://www.scopus.com/inward/record.url?scp=77955625792&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77955625792&partnerID=8YFLogxK

M3 - Article

VL - 16

SP - 887

EP - 896

JO - Molecular Vision

JF - Molecular Vision

SN - 1090-0535

ER -

Roshan M, Vijaya PH, Lavanya GR, Shama PK, Santhiya ST, Graw J et al. A novel human CRYGD mutation in a juvenile autosomal dominant cataract. Molecular Vision. 2010;16:887-896.