RIC-3 affects properties and quantity of nicotinic acetylcholine receptors via a mechanism that does not require the coiled-coil domains

J Biol Chem. 2005 Jul 29;280(30):28053-60. doi: 10.1074/jbc.M504369200. Epub 2005 Jun 2.

Abstract

Members of the RIC-3 gene family are effectors of nicotinic acetylcholine receptor (nAChR) expression in vertebrates and invertebrates. In Caenorhabditis elegans RIC-3 is needed for functional expression of multiple nAChRs, including the DEG-3/DES-2 nAChR. Effects of RIC-3 on DEG-3/DES-2 functional expression are found in vivo and following heterologous expression in Xenopus leavis oocytes. We now show that in X. leavis oocytes RIC-3 also affects the kinetics and agonist affinity properties of the DEG-3/DES-2 receptor. Because these effects are mimicked by increasing the ratio of DEG-3 subunits within DEG-3/DES-2 receptors, this suggests that RIC-3 may preferentially promote maturation of DEG-3-rich receptors. Indeed, effects of RIC-3 on functional expression of DEG-3/DES-2 positively correlate with the DEG-3 to DES-2 ratio. All RIC-3 family members have two transmembrane domains followed by one or two coiled-coil domains. Here we show that the effects of RIC-3 on functional expression and on receptor properties are mediated by the transmembrane domains and do not require the coiled-coil domains. In agreement with this, mammals express a RIC-3 transcript lacking the coiled-coil domain that is capable of promoting DEG-3/DES-2 functional expression. Last, we show that RIC-3 affects DEG-3 quantity, suggesting stabilization of receptors or receptor intermediates by RIC-3. Together our results suggest that subunit-specific interactions of RIC-3 with nAChR subunits, mediated by the transmembrane domains, are sufficient for the effects of RIC-3 on nAChR quantity and quality.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Blotting, Western
  • Caenorhabditis elegans
  • Caenorhabditis elegans Proteins / chemistry
  • Caenorhabditis elegans Proteins / physiology*
  • Cell Membrane / metabolism
  • Choline / chemistry
  • Choline / metabolism
  • Cytosol / metabolism
  • DNA, Complementary / metabolism
  • Dose-Response Relationship, Drug
  • Electrophysiology
  • Gene Deletion
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Immunoprecipitation
  • Kinetics
  • Molecular Sequence Data
  • Mutation
  • Oocytes / metabolism
  • Protein Binding
  • Protein Isoforms
  • Protein Structure, Tertiary
  • RNA, Complementary / metabolism
  • Receptors, Nicotinic / chemistry*
  • Receptors, Nicotinic / metabolism
  • Sequence Homology, Amino Acid
  • Time Factors
  • Xenopus
  • Xenopus laevis

Substances

  • Caenorhabditis elegans Proteins
  • DNA, Complementary
  • Protein Isoforms
  • RNA, Complementary
  • Receptors, Nicotinic
  • ric-3 protein, C elegans
  • Green Fluorescent Proteins
  • Choline