title: Mechanism of two novel human GJC3 missense mutations in causing non-syndromic hearing loss.
abstract: Connexins (CXs), as a component of gap junction channel, are homologous four transmembrane-domain proteins, with numerous studies confirming their auditory functions. Among a cohort of patients having incurred non-syndromic hearing loss, we identified two novel missense mutations, p.R15G and p.L23H, in the GJC3 gene encoding CX30.2/CX31.3, as causally related to hearing loss in previous study. However, the functional alteration of CX30.2/CX31.3 caused by the mutant GJC3 gene remains unknown. In this study, we compared the intracellular distribution of mutant CX30.2/CX31.3 (p.R15G and p.L23H) with the wild-type (WT) protein in HeLa cells and the effect of the mutant protein had on those cells. Analytical results indicated that p.R15G and p.L23H mutant exhibited continuous staining along apposed cell membranes in the fluorescent localization assay, which is the same with the WT. Moreover, ATP release (hemichannel function) is less in HeLa cells carrying mutant GJC3 genes than those of WT expressing cells. We believe that although p.R15G and p.L23H mutants do not decrease the trafficking of CX proteins, mutations in GJC3 genes result in a loss of hemichannel function of CX30.2/CX31.3 protein, possibly causing hearing loss. Results of this study provide a novel molecular explanation for the role of GJC3 in hearing loss.
authors.affiliation: Department of Cardiovascular Surgery, Tian-Sheng Memorial Hospital, Tong Kang, Pin-Tong, Taiwan, ROC.
authors.forename: Ching-Chyuan
authors.initials: CC
authors.lastname: Su
authors.forename: Shuan-Yow
authors.initials: SY
authors.lastname: Li
authors.forename: Yung-Chang
authors.initials: YC
authors.lastname: Yen
authors.forename: Jhih-Hao
authors.initials: JH
authors.lastname: Nian
authors.forename: Wei-Guang
authors.initials: WG
authors.lastname: Liang
authors.forename: Jiann-Jou
authors.initials: JJ
authors.lastname: Yang
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D019295:Computational Biology; D017630:Connexins / Q000737:chemistry / Q000235:genetics*; D016208:Databases, Factual; D006367:HeLa Cells; D034381:Hearing Loss / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000235:genetics*; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment
chemical_list: D017630:Connexins; C464674:GJC3 protein, human; D009419:Nerve Tissue Proteins