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Spatial encoding of cyclic AMP signaling specificity by GPCR endocytosis

Nature Chemical Biology volume 10, pages 1061–1065 (2014)Cite this article

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Abstract

G protein–coupled receptors (GPCRs) are well known to signal via cyclic AMP (cAMP) production at the plasma membrane, but it is now clear that various GPCRs also signal after internalization. Apart from its temporal impact through prolonging the cellular response, we wondered whether the endosome-initiated signal encodes any discrete spatial information. Using the β2-adrenoceptor (β2-AR) as a model, we show that endocytosis is required for the full repertoire of downstream cAMP-dependent transcriptional control. Next, we describe an orthogonal optogenetic approach to definitively establish that the location of cAMP production is indeed the critical variable determining the transcriptional response. Finally, our results suggest that this spatial encoding scheme helps cells functionally discriminate chemically distinct β2-AR ligands according to differences in their ability to promote receptor endocytosis. These findings reveal a discrete principle for achieving cellular signaling specificity based on endosome-mediated spatial encoding of intracellular second messenger production and ′location-aware′ downstream transcriptional control.

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Figure 1: Endocytosis of β2-AR is required for cAMP accumulation and transcriptional response.
Figure 2: The β2-AR transcriptional response does not require receptor recycling.
Figure 3: Transcriptional signaling is not monotonically related to net cAMP levels.
Figure 4: Localized cAMP production causes distinct transcriptional responses.

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Acknowledgements

We thank P. Brown for generously providing access to microarray facilities; P. Hegemann, H. Stenmark and T. Meyer for plasmids; D. Udwari and T. Oertner for initially suggesting the use of bPAC; and H. Bourne, B. Cheyette, R. Irannejad, B. Lobingier, A. Marley and D. Riordan for valuable discussion. These studies were supported by the National Institute on Drug Abuse of the US National Institutes of Health (DA010711 and DA012864 to M.v.Z.). N.G.T. is supported by the American Heart Association.

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Authors and Affiliations

  1. Department of Psychiatry, University of California–San Francisco, San Francisco, California, USA

    Nikoleta G Tsvetanova & Mark von Zastrow

  2. Department of Cellular and Molecular Pharmacology, University of California–San Francisco, San Francisco, California, USA

    Mark von Zastrow

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  1. Nikoleta G Tsvetanova
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Contributions

N.G.T. performed the experiments and analyzed the data. N.G.T. and M.v.Z. designed the study, interpreted the results and wrote the manuscript.

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Correspondence to Mark von Zastrow.

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The authors declare no competing financial interests.

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Tsvetanova, N., von Zastrow, M. Spatial encoding of cyclic AMP signaling specificity by GPCR endocytosis. Nat Chem Biol 10, 1061–1065 (2014). https://doi.org/10.1038/nchembio.1665

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