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Retromer terminates the generation of cAMP by internalized PTH receptors

Nature Chemical Biology volume 7, pages 278–284 (2011)Cite this article

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Abstract

The generation of cAMP by G protein–coupled receptors (GPCRs) and its termination are currently thought to occur exclusively at the plasma membrane of cells. Under existing models of receptor regulation, this signal is primarily restricted by desensitization of the receptors through their binding to β-arrestins. However, this paradigm is not consistent with recent observations that the parathyroid hormone receptor type 1 (PTHR) continues to stimulate cAMP production even after receptor internalization, as β-arrestins are known to rapidly bind and internalize activated PTHR. Here we show that binding to β-arrestin1 prolongs rather than terminates the generation of cAMP by PTHR, and that cAMP generation correlates with the persistence of arrestin–receptor complexes on endosomes. PTHR signaling is instead turned off by the retromer complex, which regulates the movement of internalized receptor from endosomes to the Golgi apparatus. Thus, binding by the retromer complex regulates the sustained generation of cAMP triggered by an internalized GPCR.

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Figure 1: Time courses of cAMP and calcium responses to PTHR.
Figure 2: Signaling and arrestin mobilization by PTH analogs.
Figure 3: Retromer terminates the PTH-mediated cAMP response.
Figure 4: Modulation of PTHR signaling by retromer.
Figure 5: Retromer and arrestin regulate PTH-mediated cAMP production in bone cells.
Figure 6: Mode of regulation of PTHR signaling by retromer and arrestin.

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Acknowledgements

This work was supported by the US National Institutes of Health award R01DK087688 (to J.-P.V.). We thank J. Bonifacino for plasmids encoding retromer subunits Vps26 and Vps29YFP and L. Traub for plasmids encoding β-arrestin1tom and β-arrestin1[I386A, V387A]tom.

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Authors and Affiliations

  1. Department of Pharmacology and Chemical Biology, Laboratory for GPCR Biology, University of Pittsburgh, School of Medicine, Pittsburgh, Pennsylvania, USA

    Timothy N Feinstein, Vanessa L Wehbi, Juan A Ardura, David S Wheeler & Jean-Pierre Vilardaga

  2. Department of Medicine, Endocrine Unit, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA

    Sebastien Ferrandon, Thomas J Gardella & Jean-Pierre Vilardaga

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  1. Timothy N Feinstein
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Contributions

T.N.F. performed most of the experiments with the support of V.L.W., J.A.A., D.S.W., S.F. and T.J.G.; J.-P.V. designed and supervised the experiments; J.-P.V. and T.N.F. analyzed the data and wrote the manuscript; all authors discussed the results and commented on the manuscript.

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Correspondence to Jean-Pierre Vilardaga.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Methods and Supplementary Figures 1–10 (PDF 1653 kb)

Supplementary Video 1

A three-dimensional reconstruction of a representative endosome labeled with Vps29YFP (blue), GFPPTHR (green) and β-arr1tom (red). (MOV 13562 kb)

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Feinstein, T., Wehbi, V., Ardura, J. et al. Retromer terminates the generation of cAMP by internalized PTH receptors. Nat Chem Biol 7, 278–284 (2011). https://doi.org/10.1038/nchembio.545

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