title: Mechanism of a novel missense mutation, p.V174M, of the human connexin31 (GJB3) in causing nonsyndromic hearing loss.
abstract: Hearing loss is the most common sensory disorder, worldwide. In a recent study, we have identified a missense mutation, p.V174M, in the connexin 31 encoded by the GJB3 gene, in a patient with nonsyndromic hearing loss. However, the functional change in the CX31V174M mutant remains unknown. This study compared the intracellular distribution and assembly of the mutant CX31V174M with that of the wild-type (WT) CX31 in HeLa cells, and it examined the effect that the mutant protein had on those cells. A fluorescent localization assay of WT CX31 showed the typical punctuate pattern of a gap junction channel between the neighboring expression cells. Conversely, the p.V174M missense mutation resulted in the accumulation of the mutant protein in the lysosomes rather than in the cytoplasmic membrane. Moreover, dye transfer experiments have also demonstrated that the CX31V174M mutant did not form functional gap junction channels, probably due to the incorrect assembly or the altered properties of the CX31 channels. In addition, we found that CX31V174M-transfection can cause cell death by MTT assay. CX31V174M co-expressed with either CX31WT or CX26WT studies, suggested the impairment of the ability of CX26WT proteins to intracellular trafficking and targeting to the plasma membrane, but did not influence the trafficking of CX31WT. Based on these findings, we suggest that the CX31V174M mutant may have an effect on the formation and function of the gap junction, and CX31V174M has a trans-dominant negative effect on the function of wild types CX26. These results provide a novel molecular explanation for the role that GJB3 plays in hearing loss.
authors.affiliation: a Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan.
authors.forename: Tung-Cheng
authors.initials: TC
authors.lastname: Li
authors.forename: Yu-Hsiang
authors.initials: YH
authors.lastname: Kuan
authors.forename: Tzu-Yu
authors.initials: TY
authors.lastname: Ko
authors.forename: Chuan
authors.initials: C
authors.lastname: Li
authors.forename: Jiann-Jou
authors.initials: JJ
authors.lastname: Yang
mesh_terms: D002470:Cell Survival; D000072259:Connexin 26; D017630:Connexins / Q000235:genetics* / Q000378:metabolism; D003638:Deafness / Q000235:genetics; D017629:Gap Junctions / Q000378:metabolism; D056726:Genetic Association Studies; D020022:Genetic Predisposition to Disease; D006367:HeLa Cells; D006801:Humans; D020125:Mutation, Missense; D021381:Protein Transport
chemical_list: D017630:Connexins; C000607042:GJB2 protein, human; D000072259:Connexin 26; C068044:GJB3 protein, human
keywords: CX31; GJB3; hearing loss; mutation; perte auditive