title: A recurrent missense mutation in GJA3 associated with autosomal dominant cataract linked to chromosome 13q.
abstract: OBJECTIVE
  To map and identify the genetic defect underlying autosomal dominant cataract segregating in a 5-generation Caucasian American family.
  
  
  METHODS
  Genomic DNA was prepared from blood leukocytes, genotyping was performed using microsatellite markers, and logarithm of the odds (LOD) scores were calculated using the LINKAGE programs. Mutation profiling was performed using direct exon cycle-sequencing and restriction fragment analysis. Protein function effects were evaluated using in silico prediction algorithms.
  
  
  RESULTS
  Significant evidence of linkage was obtained at marker D13S175 (maximum LOD score [Z(max)]=3.67; maximum recombination fraction [θ(max)]=0.04) and D13S1316 (Z(max)=2.80, θ(max)=0.0). Haplotyping indicated that the disease lay in the ~170 Kb physical interval between D13S1316 and D13S175, which contained the gene for gap-junction protein alpha-3 (GJA3) or connexin-46. Sequencing of GJA3 detected a heterozygous transition (c.130G>A) in exon-2 that resulted in gain of an Hsp92 II restriction site. Allele-specific PCR amplification and restriction analysis confirmed that the novel Hsp92 II site co-segregated with cataract in the family but was not detected in 192 normal unrelated individuals. The c.130G>A transition was predicted to result in a non-conservative substitution of valine-to-methionine at codon 44 (p.V44M) with damaging effects on protein function.
  
  
  CONCLUSIONS
  These data confirm GJA3 as one of the most frequently mutated genes that underlie autosomal dominant cataract in humans, and further emphasize the importance of connexin function in maintaining lens transparency.
authors: Department of Ophthalmology and Visual Sciences, Washington University School of Medicine, St Louis, MO, USA.
authors: Thomas M
authors: TM
authors: Bennett
authors: Alan
authors: A
authors: Shiels
mesh_terms: D000483:Alleles; D001483:Base Sequence; D016022:Case-Control Studies; D002386:Cataract / Q000235:genetics*; D025063:Chromosome Disorders / Q000235:genetics*; D002882:Chromosomes, Human, Pair 13 / Q000737:chemistry / Q000235:genetics*; D017630:Connexins / Q000235:genetics*; D004252:DNA Mutational Analysis; D005260:Female; D005799:Genes, Dominant; D008040:Genetic Linkage; D005838:Genotype; D006239:Haplotypes; D006801:Humans; D007908:Lens, Crystalline / Q000378:metabolism / Q000473:pathology; D008126:Lod Score; D008297:Male; D018895:Microsatellite Repeats; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D010375:Pedigree; D014481:United States
chemical_list: D017630:Connexins; C082917:GJA3 protein, human