Mutations in the CNGB3 gene encoding the β-subunit of the cone photoreceptor cGMP-gated channel are responsible for achromatopsia (ACHM3) linked to chromosome 8q21

• Published in: Human molecular genetics 2000-09, Vol.9 (14), p.2107-2116
• Main Authors: KOHL, S, BAUMANN, B, WISSINGER, B, BROGHAMMER, M, JÄGLE, H, SIEVING, P, KELLNER, U, SPEGAL, R, ANASTASI, M, ZRENNER, E, SHARPE, L. T
• Format: Article
• Language: English
• Subjects: ACHM3 gene; achromatopsia; Amino Acid Sequence; Animals; Biological and medical sciences; Blotting, Northern; chromosome 8; Chromosome Mapping; Chromosomes, Artificial, Yeast; Chromosomes, Human, Pair 8; CNGB3 gene; Color Vision Defects - genetics; Contig Mapping; cyclic GMP; Cyclic Nucleotide-Gated Cation Channels; DNA Mutational Analysis; DNA, Complementary - metabolism; Exons; Family Health; Female; Gene Deletion; Genetic Markers; Haplotypes; Humans; Introns; Ion Channels; Lod Score; Male; Medical sciences; Mice; Models, Genetic; Molecular Sequence Data; Mutation; Mutation, Missense; Ophthalmology; Pedigree; Photoreceptor Cells - physiology; Polymorphism, Restriction Fragment Length; Polymorphism, Single-Stranded Conformational; Retina - metabolism; Retinal Cone Photoreceptor Cells - metabolism; Reverse Transcriptase Polymerase Chain Reaction; RNA Splicing; RNA, Messenger - metabolism; Sequence Homology, Amino Acid; Sequence Tagged Sites; Tissue Distribution; Vision disorders
• ISSN: 0964-6906; 1460-2083; 1460-2083
• DOI: 10.1093/hmg/9.14.2107

Achromatopsia is an autosomal recessive disorder featuring total colour blindness, photophobia, reduced visual acuity and nystagmus. While mutations in the CNGA3 gene on chromosome 2q11 are responsible for achromatopsia in a subset of patients, previous linkage studies have localized another achromatopsia locus, ACHM3, on chromosome 8q21. Using achromatopsia families in which CNGA3 mutations have been excluded, we refined the ACHM3 locus to a 3.7 cM region enclosed by markers D8S1838 and D8S273. Two yeast artificial chromosome (YAC) contigs covering nearly the entire ACHM3 interval were constructed. Database searches with YAC content sequences identified two overlapping high throughput genomic sequencing phase (HTGS) entries which contained sequences homologous to the murine cng6 gene encoding the putative beta-subunit of the cone photoreceptor cGMP-gated channel. Using RT-PCR and RACE, we identified and cloned the human cDNA homologue, designated CNGB3, which encodes an 809 amino acid polypeptide. Northern blot analysis revealed a major transcript of approximately 4.4 kb specifically expressed in the retina. The human CNGB3 gene consists of 18 exons distributed over approximately 200 kb of genomic sequence. Analysis of the CNGB3 gene in achromats revealed six different mutations including a missense mutation (S435F), two stop codon mutations (R203X and E336X), a 1 bp and an 8 bp deletion (1148delC and 819-826del) and a putative splice site mutation of intron 13. The 1148delC mutation was identified recurrently in several families, and in total was present on 11 of 22 disease chromosomes segregating in our families.