title: Different functional consequences of two missense mutations in the GJB2 gene associated with non-syndromic hearing loss.
abstract: Mutations in the GJB2 gene, which encodes the gap junction (GJ) protein connexin26 (Cx26), are the most common cause of inherited non-syndromic hearing loss (NSHL). We identified two missense mutations, p.D46E (c.138T>G) and p.T86R (c.257C>G), of GJB2 in Korean HL families. The novel p.D46E mutation exhibited autosomal dominant inheritance, while the p.T86R mutation, which is exclusively found in Asians, segregated with an autosomal recessive pattern. Thus, we sought to elucidate the pathogenic nature of such different inherited patterns of HL. We studied protein localization and gap junction functions in cells transfected with wild-type or mutant Cx26 tagged with fluorescent proteins, which allowed visual confirmation of homozygous or heterozygous mutant GJs. The Cx26-D46E mutant was targeted to the plasma membrane, but this mutant protein failed to transfer Ca(2+) or propidium iodide intercellularly, suggesting disruption of both ionic and biochemical coupling. Heterozygous GJs also showed dysfunctional intercellular couplings and hemichannel opening, confirming the dominant-negative nature of the p.D46E mutation. The Cx26-T86R mutant protein did not form GJs, since the mutated protein was confined in the cytoplasm and not transported to the cell membrane. When Cx26-T86R was co-expressed with Cx26-WT, ionic and biochemical coupling was normal, consistent with the recessive nature of the mutation. These studies revealed distinct pathogenic mechanisms of two GJB2 mutations identified in Korean families.
authors.affiliation: Department of Biology, College of Natural Sciences, Kyungpook National University, Daegu, South Korea.
authors.forename: Soo-Young
authors.initials: SY
authors.lastname: Choi
authors.forename: Hong-Joon
authors.initials: HJ
authors.lastname: Park
authors.forename: Kyu Yup
authors.initials: KY
authors.lastname: Lee
authors.forename: Emilie Hoang
authors.initials: EH
authors.lastname: Dinh
authors.forename: Qing
authors.initials: Q
authors.lastname: Chang
authors.forename: Shoab
authors.initials: S
authors.lastname: Ahmad
authors.forename: Sang Heun
authors.initials: SH
authors.lastname: Lee
authors.forename: Jinwoong
authors.initials: J
authors.lastname: Bok
authors.forename: Xi
authors.initials: X
authors.lastname: Lin
authors.forename: Un-Kyung
authors.initials: UK
authors.lastname: Kim
mesh_terms: D002460:Cell Line; D000072259:Connexin 26; D017630:Connexins / Q000235:genetics*; D005192:Family Health; D017629:Gap Junctions / Q000235:genetics; D005799:Genes, Dominant; D005808:Genes, Recessive; D034381:Hearing Loss / Q000235:genetics*; D006801:Humans; D007723:Korea; D020125:Mutation, Missense; D021381:Protein Transport; D014162:Transfection
chemical_list: D017630:Connexins; C000607042:GJB2 protein, human; D000072259:Connexin 26