title: Properties of two cataract-associated mutations located in the NH2 terminus of connexin 46.
abstract: Mutations in connexin 46 are associated with congenital cataracts. The purpose of this project was to characterize cellular and functional properties of two congenital cataract-associated mutations located in the NH2 terminus of connexin 46: Cx46D3Y and Cx46L11S, which we found localized to gap junctional plaques like wild-type Cx46 in transfected HeLa cells. Dual two-microelectrode-voltage-clamp studies of Xenopus oocyte pairs injected with wild-type or mutant rat Cx46 showed that oocyte pairs injected with D3Y or L11S cRNA failed to induce gap junctional coupling, whereas oocyte pairs injected with Cx46 showed high levels of coupling. D3Y, but not L11S, functionally paired with wild-type Cx46. To determine whether coexpression of D3Y or L11S affected the junctional conductance produced by wild-type lens connexins, we studied pairs of oocytes coinjected with equal amounts of mutant and wild-type connexin cRNA. Expression of D3Y or L11S almost completely abolished gap junctional coupling induced by Cx46. In contrast, expression of D3Y or L11S failed to inhibit junctional conductance induced by Cx50. To examine effects of the D3Y and L11S mutations on hemichannel activity, hemichannel currents were measured in connexin cRNA-injected oocytes. Oocytes expressing D3Y exhibited reduced hemichannel activity as well as alterations in voltage gating and charge selectivity while oocytes expressing L11S showed no hemichannel activity. Moreover, coexpression of mutant with wild-type Cx50 or Cx46 gave rise to hemichannels with distinct electrophysiological properties, suggesting that the mutant connexins were forming heteromeric channels with wild-type connexins. These data suggest D3Y and L11S cause cataracts by similar but not identical mechanisms.
authors.affiliation: Department of Physiology and Biophysics, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064, USA.
authors.forename: Jun-Jie
authors.initials: JJ
authors.lastname: Tong
authors.forename: Bonnie C H
authors.initials: BC
authors.lastname: Sohn
authors.forename: Anh
authors.initials: A
authors.lastname: Lam
authors.forename: D Eric
authors.initials: DE
authors.lastname: Walters
authors.forename: Barbara M
authors.initials: BM
authors.lastname: Vertel
authors.forename: Lisa
authors.initials: L
authors.lastname: Ebihara
mesh_terms: D000818:Animals; D002386:Cataract / Q000151:congenital / Q000235:genetics* / Q000473:pathology; D017630:Connexins / Q000235:genetics* / Q000378:metabolism; D017629:Gap Junctions / Q000378:metabolism / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D015640:Ion Channel Gating; D008564:Membrane Potentials; D020125:Mutation, Missense; D018408:Patch-Clamp Techniques; D017434:Protein Structure, Tertiary; D051381:Rats; D040681:Structural Homology, Protein; D014982:Xenopus laevis
chemical_list: D017630:Connexins; C082917:GJA3 protein, human
keywords: cataract; connexin; gap junction; intercellular communication