An N-terminal region of Caenorhabditis elegans RGS proteins EGL-10 and EAT-16 directs inhibition of G(alpha)o versus G(alpha)q signaling

J Biol Chem. 2002 Dec 6;277(49):47004-13. doi: 10.1074/jbc.M208186200. Epub 2002 Sep 26.

Abstract

Regulator of G protein signaling (RGS) proteins contain an RGS domain that inhibits G(alpha) signaling by activating G(alpha) GTPase activity. Certain RGS proteins also contain a Ggamma-like (GGL) domain and a poorly characterized but conserved N-terminal region. We assessed the functions of these subregions in the Caenorhabditis elegans RGS proteins EGL-10 and EAT-16, which selectively inhibit GOA-1 (G(alpha)(o)) and EGL-30 (G(alpha)(q)), respectively. Using transgenes in C. elegans, we expressed EGL-10, EAT-16, their subregions, or EGL-10/EAT-16 chimeras. The chimeras showed that the GGL/RGS region of either protein can act on either GOA-1 or EGL-30 and that a key factor determining G(alpha) target selectivity is the manner in which the N-terminal and GGL/RGS regions are linked. We also found that coexpressing N-terminal and GGL/RGS fragments of EGL-10 gave full EGL-10 activity, whereas either fragment alone gave little activity. Biochemical analysis showed that coexpressing the two fragments caused both to increase in abundance and also caused the GGL/RGS fragment to move to the membrane, where the N-terminal fragment is localized. By coimmunoprecipitation, we found that the N-terminal fragment complexes with the C-terminal fragment and its associated Gbeta subunit, GPB-2. We conclude that the N-terminal region directs inhibition of G(alpha) signaling by forming a complex with the GGL/RGS region and affecting its stability, membrane localization, and G(alpha) target specificity.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Animals, Genetically Modified
  • Blotting, Western
  • Caenorhabditis elegans / chemistry*
  • Caenorhabditis elegans Proteins*
  • Cell Membrane / metabolism
  • Chromosomes / ultrastructure
  • Epitopes
  • GTP-Binding Protein Regulators*
  • Helminth Proteins / chemistry*
  • Heterotrimeric GTP-Binding Proteins / chemistry*
  • Heterotrimeric GTP-Binding Proteins / metabolism
  • Immunoblotting
  • Models, Biological
  • Molecular Sequence Data
  • Mutation
  • Plasmids / metabolism
  • Promoter Regions, Genetic
  • Protein Binding
  • Protein Structure, Tertiary
  • Proteins / metabolism
  • RGS Proteins / chemistry*
  • Sequence Homology, Amino Acid
  • Signal Transduction
  • Time Factors
  • Transgenes

Substances

  • Caenorhabditis elegans Proteins
  • EAT-16 protein, C elegans
  • EGL-10 protein, C elegans
  • Epitopes
  • GTP-Binding Protein Regulators
  • Helminth Proteins
  • Proteins
  • RGS Proteins
  • Heterotrimeric GTP-Binding Proteins