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Spontaneous receptor-independent heterotrimeric G-protein signalling in an RGS mutant

Nature Cell Biology volume 5, pages 231–235 (2003)Cite this article

Abstract

Tripartite G-protein-coupled receptors (GPCRs) represent one of the largest groups of signal transducers, transmitting signals from hormones, neuropeptides, odorants, food and light1,2. Ligand-bound receptors catalyse GDP/GTP exchange on the G-protein α-subunit (Gα), leading to α-GTP separation from the βγ subunits and pathway activation. Activating mutations in the receptors or G proteins underlie many human diseases, including some cancers, dwarfism and premature puberty. Regulators of G-protein signalling (RGS proteins) are known to modulate the level and duration of ligand-induced signalling by accelerating the intrinsic GTPase activity of the Gα subunit, and thus reformation of the inactive GDP-bound Gα3,4,5. Here we find that even in the absence of receptor, mutation of the RGS family member Sst2 (refs 6–9) permits spontaneous activation of the G-protein-coupled mating pathway in Saccharomyces cerevisiae at levels normally seen only in the presence of ligand. Our work demonstrates the occurence of spontaneous tripartite G-protein signalling in vivo and identifies a requirement for RGS proteins in preventing such receptor-independent activation.

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Figure 1: sst2Δ mating morphologies are not the result of mating-type switching.
Figure 2: sst2Δ morphologies require the mating pathway, but not receptor activation.
Figure 3: Transcriptional induction in sst2Δ cells at levels seen in mating wild-type cells.
Figure 4: sst2Δ mating morphologies are the result of Gα activation.

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Acknowledgements

We thank J. Thorner for experimental advice, S. Avery for pSVA12 and 13, H. Dohlman for the Sst2 antibody and pGPA1G302S, I. Herskowitz for the HMLa HMRa strain, S. Reed and D. Stone for pGPA1G322E and R, C. Beh and J. Rine for the FUS1:GFP plasmid, J. Dohmen for pPW66R, and H. Nolla of the UC Berkeley Cancer Research Laboratory Flow Cytometry Facility for help with FACS. We thank C. Boone, H. Bourne, J. Rine, J. Thorner, M. Albers, K. Karbstein, K. Kozminski, G. Manning and the Drubin Laboratory for insightful discussions and comments on the manuscript. This work was supported by NIH grants GM20291 to D.E.S. and GM50399 to D.G.D.

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  1. Department of Molecular and Cell Biology, University of California Berkeley, 401 Barker Hall, Berkeley, 94720-3202, CA, USA

    Daria E. Siekhaus & David G. Drubin

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Correspondence to David G. Drubin.

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Figure S1 Calibration of FACS data. (PDF 274 kb)

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Siekhaus, D., Drubin, D. Spontaneous receptor-independent heterotrimeric G-protein signalling in an RGS mutant. Nat Cell Biol 5, 231–235 (2003). https://doi.org/10.1038/ncb941

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