Mutant cochlear connexins associated with deafness

Summary

Principal Investigator: Guillermo Altenberg
Abstract: Gap junctions are aqueous channels permeable to ions and hydrophilic molecules of Mr < 1,000. Each gapjunctional channel is formed by head-to-head association of two connexons (connexin hexamers), one from each of the adjacent cells. The cochlea (responsible for mechano-transduction of sound waves into electrical impulses) has a highly-developed gap-junctional network that is essential for hearing. In most cells of the normal cochlea gap-junctional channels are heteromeric assemblies formed by the connexins Cx26 and Cx30. Infant deafness due to mutations of Cx26 is very common and our long-term objective is to elucidate at the molecular level the mechanisms by which mutations of connexins cause deafness. The properties of wild-type Cx26, Cx30 and heteromeric Cx26/Cx30 connexons, and the effects of mutations that cause deafness on the properties of heteromeric connexons that include wild-type connexins, are poorly understood. This is in part due to the fact that the few groups that have carried out functional experiments on Cx26 and Cx30 mutants work with complex systems, performing measurements that depend on the gapjunctional communication between two neighboring cells. This kind of experiments is essential to understand the effects of the mutations, but cannot fully address the molecular mechanism of the alterations. The specific aims of this proposal are: 1) to develop an expression/purification/reconstitution system that yields large amounts of functional wild-type and mutant Cx26 and Cx30, and 2) to test the usefulness of the purified connexon system to determine the functional properties of connexons formed by wild-type and mutant connexins. We will adapt the methodology that we developed recently for Cx43, which should allow us to obtain large amounts of functional Cx26 and Cx30, and connexin mutants. We will study the permeability properties of wild-type and mutant homomeric Cx26 and Cx30 connexons and heteromeric Cx26/Cx30 connexons formed by wild-type connexins as well as connexons containing Cx26 or Cx30 mutants. An integrative approach comparing the properties of gap- junctional channels and connexons will allow us to determine whether specific mutations alter gap-junctional communication at the level of the gap-junctional channel (e.g., docking between GJH) or connexon (e.g., non-permeable connexons).
Funding Period: 2005-09-01 - 2007-03-31
more information: NIH RePORT

Top Publications

  1. ncbi Mutations of connexin 26 at position 75 and dominant deafness: essential role of arginine for the generation of functional gap-junctional channels
    Yanqin Deng
    Department of Neuroscience and Cell Biology, and the Sealy Center for Structural Biology and Molecular Biophysics, The University of Texas Medical Branch, Galveston, TX 77555 0437, United States
    Hear Res 220:87-94. 2006
  2. pmc Change in permeant size selectivity by phosphorylation of connexin 43 gap-junctional hemichannels by PKC
    Xiaoyong Bao
    Department of Neuroscience and Cell Biology, University of Texas Medical Branch, Galveston, TX 77555 0437, USA
    Proc Natl Acad Sci U S A 104:4919-24. 2007
  3. pmc Molecular modeling and mutagenesis of gap junction channels
    Julio A Kovacs
    Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037, USA
    Prog Biophys Mol Biol 94:15-28. 2007

Detail Information

Publications3

  1. ncbi Mutations of connexin 26 at position 75 and dominant deafness: essential role of arginine for the generation of functional gap-junctional channels
    Yanqin Deng
    Department of Neuroscience and Cell Biology, and the Sealy Center for Structural Biology and Molecular Biophysics, The University of Texas Medical Branch, Galveston, TX 77555 0437, United States
    Hear Res 220:87-94. 2006
    ..Our results show that the arginine located at position 75 of connexin 26 is essential for function, and cannot be replaced by other residues...
  2. pmc Change in permeant size selectivity by phosphorylation of connexin 43 gap-junctional hemichannels by PKC
    Xiaoyong Bao
    Department of Neuroscience and Cell Biology, University of Texas Medical Branch, Galveston, TX 77555 0437, USA
    Proc Natl Acad Sci U S A 104:4919-24. 2007
    ..These results indicate that phosphorylation of Cx43 by PKC alters the hemichannel size selectivity and explain why PKC activity affects dye transfer between cells without consistent effects on electrical communication...
  3. pmc Molecular modeling and mutagenesis of gap junction channels
    Julio A Kovacs
    Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037, USA
    Prog Biophys Mol Biol 94:15-28. 2007
    ..In spite of this substantial progress in understanding the structural biology of gap junction channels, an experimentally determined structure at atomic resolution will be essential to confirm these concepts...