G12/13 is activated by acute tethered agonist exposure in the adhesion GPCR ADGRL3

Nat Chem Biol. 2020 Dec;16(12):1343-1350. doi: 10.1038/s41589-020-0617-7. Epub 2020 Aug 10.

Abstract

The adhesion G-protein-coupled receptor (GPCR) latrophilin 3 (ADGRL3) has been associated with increased risk of attention deficit hyperactivity disorder (ADHD) and substance use in human genetic studies. Knockdown in multiple species leads to hyperlocomotion and altered dopamine signaling. Thus, ADGRL3 is a potential target for treatment of neuropsychiatric disorders that involve dopamine dysfunction, but its basic signaling properties are poorly understood. Identification of adhesion GPCR signaling partners has been limited by a lack of tools to acutely activate these receptors in living cells. Here, we design a novel acute activation strategy to characterize ADGRL3 signaling by engineering a receptor construct in which we could trigger acute activation enzymatically. Using this assay, we found that ADGRL3 signals through G12/G13 and Gq, with G12/13 the most robustly activated. Gα12/13 is a new player in ADGRL3 biology, opening up unexplored roles for ADGRL3 in the brain. Our methodological advancements should be broadly useful in adhesion GPCR research.

Publication types

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

MeSH terms

  • Activating Transcription Factor 6 / agonists
  • Activating Transcription Factor 6 / chemistry
  • Activating Transcription Factor 6 / genetics
  • Activating Transcription Factor 6 / metabolism*
  • Animals
  • Arrestin / chemistry
  • Arrestin / genetics
  • Arrestin / metabolism
  • CRISPR-Cas Systems
  • Cell Engineering
  • GTP-Binding Protein alpha Subunits, G12-G13 / chemistry
  • GTP-Binding Protein alpha Subunits, G12-G13 / genetics
  • GTP-Binding Protein alpha Subunits, G12-G13 / metabolism*
  • GTP-Binding Protein alpha Subunits, Gq-G11 / chemistry
  • GTP-Binding Protein alpha Subunits, Gq-G11 / genetics
  • GTP-Binding Protein alpha Subunits, Gq-G11 / metabolism
  • Gene Expression
  • HEK293 Cells
  • Humans
  • Kinetics
  • Mice
  • Mitogen-Activated Protein Kinase 1 / chemistry
  • Mitogen-Activated Protein Kinase 1 / genetics
  • Mitogen-Activated Protein Kinase 1 / metabolism
  • Mitogen-Activated Protein Kinase 3 / chemistry
  • Mitogen-Activated Protein Kinase 3 / genetics
  • Mitogen-Activated Protein Kinase 3 / metabolism
  • Peptides / chemistry
  • Peptides / metabolism*
  • Peptides / pharmacology
  • Protein Binding
  • Receptors, G-Protein-Coupled / chemistry
  • Receptors, G-Protein-Coupled / genetics
  • Receptors, G-Protein-Coupled / metabolism*
  • Receptors, Peptide / chemistry
  • Receptors, Peptide / genetics
  • Receptors, Peptide / metabolism*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Signal Transduction

Substances

  • ADGRL3 protein, human
  • ATF6B protein, human
  • Activating Transcription Factor 6
  • Arrestin
  • Peptides
  • Receptors, G-Protein-Coupled
  • Receptors, Peptide
  • Recombinant Proteins
  • MAPK1 protein, human
  • MAPK3 protein, human
  • Mitogen-Activated Protein Kinase 1
  • Mitogen-Activated Protein Kinase 3
  • GTP-Binding Protein alpha Subunits, G12-G13
  • GTP-Binding Protein alpha Subunits, Gq-G11