Chemical Genetics Reveals an RGS/G-protein Role in the Action of a Compound

Overview

Journal PLoS Genet

Specialty Genetics

Date 2006 May 10

PMID 16683034

Citations 16

Authors

Kevin FitzGerald

Svetlana Tertyshnikova

Lisa Moore

Lynn Bjerke

Ben Burley

Jian Cao

Pamela Carroll

Robert Choy

Steve Doberstein

Yves Dubaquie

Yvonne Franke

Jenny Kopczynski

Hendrik Korswagen

Stanley R Krystek

Nicholas J Lodge

Ronald Plasterk

John Starrett

Terry Stouch

George Thalody

Honey Wayne

Alexander van der Linden

Yongmei Zhang

Stephen G Walker

Mark Cockett

Judi Wardwell-Swanson

Petra Ross-Macdonald

Rachel M Kindt

Affiliations

Soon will be listed here.

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.

Citing Articles

New insights into the pharmacological inhibition of SRF activity: Key inhibitory targets and mechanisms.

Wong D, Qiu H Vascul Pharmacol. 2024; 157():107443.

PMID: 39586415 PMC: 11648470. DOI: 10.1016/j.vph.2024.107443.

RGS2 and female common diseases: a guard of women's health.

Xu Q, Yao M, Tang C J Transl Med. 2023; 21(1):583.

PMID: 37649067 PMC: 10469436. DOI: 10.1186/s12967-023-04462-3.

Antiaging agents: safe interventions to slow aging and healthy life span extension.

Liu J Nat Prod Bioprospect. 2022; 12(1):18.

PMID: 35534591 PMC: 9086005. DOI: 10.1007/s13659-022-00339-y.

Acute and 28-days subacute toxicity studies of Gαq-RGS2 signaling inhibitor.

Beladiya J, Mehta A Lab Anim Res. 2021; 37(1):17.

PMID: 34311782 PMC: 8314442. DOI: 10.1186/s42826-021-00093-1.

Regulator of G-Protein Signaling (RGS) Protein Modulation of Opioid Receptor Signaling as a Potential Target for Pain Management.

Senese N, Kandasamy R, Kochan K, Traynor J Front Mol Neurosci. 2020; 13:5.

PMID: 32038168 PMC: 6992652. DOI: 10.3389/fnmol.2020.00005.

References

Lazareno S, Birdsall N . Pharmacological characterization of acetylcholine-stimulated [35S]-GTP gamma S binding mediated by human muscarinic m1-m4 receptors: antagonist studies. Br J Pharmacol. 1993; 109(4):1120-7. PMC: 2175752. DOI: 10.1111/j.1476-5381.1993.tb13738.x. View

Wall M, Coleman D, Lee E, Posner B, GILMAN A, Sprang S . The structure of the G protein heterotrimer Gi alpha 1 beta 1 gamma 2. Cell. 1995; 83(6):1047-58. DOI: 10.1016/0092-8674(95)90220-1. View

Wess J . G-protein-coupled receptors: molecular mechanisms involved in receptor activation and selectivity of G-protein recognition. FASEB J. 1997; 11(5):346-54. View

Chen W, Patikoglou G, Koelle M, Sternberg P . Antagonism between G(o)alpha and G(q)alpha in Caenorhabditis elegans: the RGS protein EAT-16 is necessary for G(o)alpha signaling and regulates G(q)alpha activity. Genes Dev. 1999; 13(14):1780-93. PMC: 316886. DOI: 10.1101/gad.13.14.1780. View

Kaslow H, Lim L, Moss J, Lesikar D . Structure-activity analysis of the activation of pertussis toxin. Biochemistry. 1987; 26(1):123-7. DOI: 10.1021/bi00375a018. View

Malkowicz S, Wein A, Elbadawi A, Van Arsdalen K, Ruggieri M, Levin R . Acute biochemical and functional alterations in the partially obstructed rabbit urinary bladder. J Urol. 1986; 136(6):1324-9. DOI: 10.1016/s0022-5347(17)45331-6. View

Stone D, Reed S . G protein mutations that alter the pheromone response in Saccharomyces cerevisiae. Mol Cell Biol. 1990; 10(9):4439-46. PMC: 361029. DOI: 10.1128/mcb.10.9.4439-4446.1990. View

Penn R, Pronin A, Benovic J . Regulation of G protein-coupled receptor kinases. Trends Cardiovasc Med. 2001; 10(2):81-9. DOI: 10.1016/s1050-1738(00)00053-0. View

Minassian V, Drutz H, Al-Badr A . Urinary incontinence as a worldwide problem. Int J Gynaecol Obstet. 2003; 82(3):327-38. DOI: 10.1016/s0020-7292(03)00220-0. View

10.

Blinder D, Jenness D . Regulation of postreceptor signaling in the pheromone response pathway of Saccharomyces cerevisiae. Mol Cell Biol. 1989; 9(9):3720-6. PMC: 362432. DOI: 10.1128/mcb.9.9.3720-3726.1989. View

11.

Khawaja X, Liang J, Saugstad J, Jones P, Harnish S, Conn P . Immunohistochemical distribution of RGS7 protein and cellular selectivity in colocalizing with Galphaq proteins in the adult rat brain. J Neurochem. 1999; 72(1):174-84. DOI: 10.1046/j.1471-4159.1999.0720174.x. View

12.

Zhou J, Toth P, Miller R . Direct interactions between the heterotrimeric G protein subunit G beta 5 and the G protein gamma subunit-like domain-containing regulator of G protein signaling 11: gain of function of cyan fluorescent protein-tagged G gamma 3. J Pharmacol Exp Ther. 2003; 305(2):460-6. DOI: 10.1124/jpet.102.048637. View

13.

Klockner U, Trieschmann U, Isenberg G . Pharmacological modulation of calcium and potassium channels in isolated vascular smooth muscle cells. Arzneimittelforschung. 1989; 39(1A):120-6. View

14.

Patikoglou G, Koelle M . 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; 277(49):47004-13. DOI: 10.1074/jbc.M208186200. View

15.

Leavitt L, Macaluso C, Kim K, Martin N, Dumont M . Dominant negative mutations in the alpha-factor receptor, a G protein-coupled receptor encoded by the STE2 gene of the yeast Saccharomyces cerevisiae. Mol Gen Genet. 1999; 261(6):917-32. DOI: 10.1007/s004380051039. View

16.

Brundage L, Avery L, Katz A, Kim U, Mendel J, Sternberg P . Mutations in a C. elegans Gqalpha gene disrupt movement, egg laying, and viability. Neuron. 1996; 16(5):999-1009. PMC: 4444781. DOI: 10.1016/s0896-6273(00)80123-3. View

17.

DiBello P, Garrison T, Apanovitch D, Hoffman G, Shuey D, Mason K . Selective uncoupling of RGS action by a single point mutation in the G protein alpha-subunit. J Biol Chem. 1998; 273(10):5780-4. DOI: 10.1074/jbc.273.10.5780. View

18.

Li E, Meldrum E, Stratton H, Stone D . Substitutions in the pheromone-responsive Gbeta protein of Saccharomyces cerevisiae confer a defect in recovery from pheromone treatment. Genetics. 1998; 148(3):947-61. PMC: 1460049. DOI: 10.1093/genetics/148.3.947. View

19.

De Backer M, Gommeren W, Moereels H, Nobels G, Van Gompel P, Leysen J . Genomic cloning, heterologous expression and pharmacological characterization of a human histamine H1 receptor. Biochem Biophys Res Commun. 1993; 197(3):1601-8. DOI: 10.1006/bbrc.1993.2662. View

20.

Kwok P . High-throughput genotyping assay approaches. Pharmacogenomics. 2001; 1(1):95-100. DOI: 10.1517/14622416.1.1.95. View