Spatial encoding of cyclic AMP signaling specificity by GPCR endocytosis

Nat Chem Biol. 2014 Dec;10(12):1061-5. doi: 10.1038/nchembio.1665. Epub 2014 Nov 2.


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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenylyl Cyclases / genetics*
  • Adenylyl Cyclases / metabolism
  • Adrenergic beta-2 Receptor Antagonists / pharmacology
  • Cell Membrane / drug effects
  • Cell Membrane / metabolism*
  • Cyclic AMP / metabolism*
  • Dynamins / antagonists & inhibitors
  • Dynamins / genetics
  • Dynamins / metabolism
  • Endocytosis
  • Endosomes / drug effects
  • Endosomes / metabolism*
  • Gene Expression Regulation
  • HEK293 Cells
  • Humans
  • Hydrazones / pharmacology
  • Naphthols / pharmacology
  • Oligonucleotide Array Sequence Analysis
  • Phosphorylation
  • Protein Transport
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • Receptors, Adrenergic, beta-2 / genetics*
  • Receptors, Adrenergic, beta-2 / metabolism
  • Signal Transduction
  • Transcription, Genetic


  • ADRB2 protein, human
  • Adrenergic beta-2 Receptor Antagonists
  • Hydrazones
  • Naphthols
  • RNA, Small Interfering
  • Receptors, Adrenergic, beta-2
  • dyngo-4a
  • Cyclic AMP
  • Dynamins
  • Adenylyl Cyclases