Chemical genetics reveals an RGS/G-protein role in the action of a compound

PLoS Genet. 2006 Apr;2(4):e57. doi: 10.1371/journal.pgen.0020057. Epub 2006 Apr 21.

Abstract

We report here on a chemical genetic screen designed to address the mechanism of action of a small molecule. Small molecules that were active in models of urinary incontinence were tested on the nematode Caenorhabditis elegans, and the resulting phenotypes were used as readouts in a genetic screen to identify possible molecular targets. The mutations giving resistance to compound were found to affect members of the RGS protein/G-protein complex. Studies in mammalian systems confirmed that the small molecules inhibit muscarinic G-protein coupled receptor (GPCR) signaling involving G-alphaq (G-protein alpha subunit). Our studies suggest that the small molecules act at the level of the RGS/G-alphaq signaling complex, and define new mutations in both RGS and G-alphaq, including a unique hypo-adapation allele of G-alphaq. These findings suggest that therapeutics targeted to downstream components of GPCR signaling may be effective for treatment of diseases involving inappropriate receptor activation.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Caenorhabditis elegans / drug effects
  • Caenorhabditis elegans / genetics*
  • Caenorhabditis elegans Proteins / genetics*
  • Caenorhabditis elegans Proteins / metabolism
  • Calcium / metabolism
  • Cell Line
  • Drug Evaluation, Preclinical
  • Female
  • GTP-Binding Protein alpha Subunits / genetics
  • GTP-Binding Protein alpha Subunits / metabolism*
  • Humans
  • RGS Proteins / genetics
  • RGS Proteins / metabolism*
  • Radioligand Assay
  • Rats
  • Rats, Sprague-Dawley
  • Receptors, G-Protein-Coupled / genetics
  • Receptors, G-Protein-Coupled / metabolism*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism
  • Triazoles / pharmacology*
  • Urinary Bladder / drug effects
  • Urinary Bladder / metabolism

Substances

  • Caenorhabditis elegans Proteins
  • GTP-Binding Protein alpha Subunits
  • RGS Proteins
  • Receptors, G-Protein-Coupled
  • Saccharomyces cerevisiae Proteins
  • Triazoles
  • Calcium