Activity-based, bioorthogonal imaging of phospholipase D reveals spatiotemporal dynamics of GPCR-Gq signaling. Issue 1 (20th January 2022) - Liang, Dongjun
Summary: Canonically, G-protein-coupled receptor (GPCR) signaling is transient and confined to the plasma membrane (PM). Deviating from this paradigm, the parathyroid hormone receptor (PTHR1) stimulates sustained Gs signaling at endosomes. In addition to Gs, PTHR1 activates Gq signaling; yet, in contrast to the PTHR1-Gs pathway, the spatiotemporal dynamics of the Gq branch of PTHR1 signaling and its relationship to Gs signaling remain largely ill defined. Recognizing that a downstream consequence of Gq signaling is the activation of phospholipase D (PLD) enzymes, we leverage activity-based, bioorthogonal imaging tools for PLD signaling to visualize and quantify the Gq branch of PTHR1 signaling. We establish that PTHR1-Gq signaling is short lived, exclusively at the PM, and antagonized by PTHR1 endocytosis. Our data support a model wherein Gq and Gs compete for ligand-bound receptors at the PM and more broadly highlight the utility of bioorthogonal tools for imaging PLDs as probes to visualize GPCR-Gq signaling. Graphical abstract: Highlights: RT-IMPACT is an activity-based method to image phospholipase D signaling RT-IMPACT specifically visualizes Gq signaling downstream of PTHR1 activation PTHR1 signals via Gq transiently and at the plasma membrane, in contrast to Gs Bioorthogonal tools for imaging PLDs are useful probes to image GPCR-Gq signaling Abstract : The parathyroid hormone receptor (PTHR1) signals via both Gs and Gq, with the former occurring, non-classically, on endosomes. Here, Liang et al. apply a bioorthogonal method for visualizing phospholipase D signaling to reveal that PTHR1-Gq signaling is transient and localized exclusively at the plasma membrane.