Detection of Concordance between Transcriptional Levels of GPCRs and Receptor-Activity-Modifying Proteins

Shahar Barbash; Torbjörn Persson; Emily Lorenzen; Manija A Kazmi; Thomas Huber; Thomas P Sakmar

doi:10.1016/j.isci.2018.12.024

Detection of Concordance between Transcriptional Levels of GPCRs and Receptor-Activity-Modifying Proteins

iScience. 2019 Jan 25:11:366-374. doi: 10.1016/j.isci.2018.12.024. Epub 2018 Dec 31.

Authors

Shahar Barbash¹, Torbjörn Persson², Emily Lorenzen¹, Manija A Kazmi¹, Thomas Huber³, Thomas P Sakmar⁴

Affiliations

¹ Laboratory of Chemical Biology and Signal Transduction, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.
² Laboratory of Chemical Biology and Signal Transduction, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA; Department of Neurobiology, Care Sciences and Society, Division for Neurogeriatrics, Center for Alzheimer Research, Karolinska Institutet, 141 57 Huddinge, Sweden.
³ Laboratory of Chemical Biology and Signal Transduction, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA. Electronic address: hubert@rockefeller.edu.
⁴ Laboratory of Chemical Biology and Signal Transduction, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA; Department of Neurobiology, Care Sciences and Society, Division for Neurogeriatrics, Center for Alzheimer Research, Karolinska Institutet, 141 57 Huddinge, Sweden. Electronic address: sakmar@rockefeller.edu.

Abstract

A recent phylogenetic analysis showed global co-evolution of G protein-coupled receptors (GPCRs) and receptor-activity-modifying proteins (RAMPs) suggesting global interactions between these two protein families. Experimental validation of these findings is challenging because in humans whereas there are only three genes encoding RAMPs, there are about 800 genes encoding GPCRs. Here, we report an experimental approach to evaluate GPCR-RAMP interactions. As a proof-of-concept experiment, we over-expressed RAMP2 in HEK293T cells and evaluated the effect on the transcriptional levels of 14 representative GPCRs that were selected based on the earlier phylogenetic analysis. We utilized a multiplexed error-correcting fluorescence in situ hybridization (MERFISH) method to detect message levels for individual GPCRs in single cells. The MERFISH results showed changes in GPCR message levels with RAMP2 over-expression in a concordant pattern that was predicted by the earlier phylogenetic analysis. These results provide additional evidence that GPCR-RAMP interactions are more widespread than previously appreciated and that these interactions have functional consequences.

Keywords: Biological Sciences; Cell Biology; Molecular Biology.