Allosteric ligands control the activation of a class C GPCR heterodimer by acting at the transmembrane interface

  1. Lei Liu
  2. Zhiran Fan
  3. Xavier Rovira  Is a corresponding author
  4. Li Xue
  5. Salomé Roux
  6. Isabelle Brabet
  7. Mingxia Xin
  8. Jean-Philippe Pin  Is a corresponding author
  9. Philippe Rondard  Is a corresponding author
  10. Jianfeng Liu  Is a corresponding author
  1. Huazhong University of Science and Technology, China
  2. Spanish National Research Council, Spain
  3. University of Montpellier, CNRS, INSERM, France

Abstract

G protein-coupled receptors (GPCRs) are among the most promising drug targets. They often form homo- and heterodimers with allosteric cross-talk between receptor entities, which contributes to fine tuning of transmembrane signaling. Specifically controlling the activity of GPCR dimers with ligands is a good approach to clarify their physiological roles and to validate them as drug targets. Here, we examined the mode of action of positive allosteric modulators (PAMs) that bind at the interface of the transmembrane domains of the heterodimeric GABAB receptor. Our site-directed mutagenesis results show that mutations of this interface impact the function of the three PAM tested. The data support the inference that they act at the active interface between both transmembrane domains, the binding site involving residues of the TM6s of the GABAB1 and the GABAB2 subunit. Importantly, the agonist activity of these PAMs involves a key region in the central core of the GABAB2 transmembrane domain, which also controls the constitutive activity of the GABAB receptor. This region corresponds to the sodium ion binding site in class A GPCRs that controls the basal state of the receptors. Overall, these data reveal the possibility of developing allosteric compounds able to specifically modulate the activity of GPCR homo- and heterodimers by acting at their transmembrane interface.

Data availability

Figure 2- Source Data 1 contain the numerical data used to generate the figures;Figure 3 - Source Data 1 contain the numerical data used to generate the figures;Figure 4 - Source Data 1 contain the numerical data used to generate the figures;Figure 5 - Source Data 1 contain the numerical data used to generate the figures.

Article and author information

Author details

  1. Lei Liu

    Cellular Signaling Laboratory, International Research Center for Sensory Biology and Technology of MOST, Huazhong University of Science and Technology, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9824-9570
  2. Zhiran Fan

    Cellular Signaling Laboratory, International Research Center for Sensory Biology and Technology of MOST, Huazhong University of Science and Technology, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9543-1211
  3. Xavier Rovira

    MCS, Laboratory of Medicinal Chemistry, Institute for Advanced Chemistry of Catalonia (IQAC-CSIC), Spanish National Research Council, Barcelona, Spain
    For correspondence
    xavier.rovira@cid.csic.es
    Competing interests
    The authors declare that no competing interests exist.
  4. Li Xue

    Cellular Signaling Laboratory, International Research Center for Sensory Biology and Technology of MOST, Huazhong University of Science and Technology, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
  5. Salomé Roux

    Institut de Génomique Fonctionnelle, University of Montpellier, CNRS, INSERM, Montpellier, France
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6106-4863
  6. Isabelle Brabet

    Institut de Génomique Fonctionnelle, University of Montpellier, CNRS, INSERM, Montpellier, France
    Competing interests
    The authors declare that no competing interests exist.
  7. Mingxia Xin

    Cellular Signaling Laboratory, International Research Center for Sensory Biology and Technology of MOST, Huazhong University of Science and Technology, Wuhan, China
    Competing interests
    The authors declare that no competing interests exist.
  8. Jean-Philippe Pin

    Institut de Génomique Fonctionnelle, University of Montpellier, CNRS, INSERM, Montpellier, France
    For correspondence
    jean-philippe.pin@igf.cnrs.fr
    Competing interests
    The authors declare that no competing interests exist.
  9. Philippe Rondard

    Institut de Génomique Fonctionnelle, University of Montpellier, CNRS, INSERM, Montpellier, France
    For correspondence
    philippe.rondard@igf.cnrs.fr
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1134-2738
  10. Jianfeng Liu

    Cellular Signaling Laboratory, International Research Center for Sensory Biology and Technology of MOST, Huazhong University of Science and Technology, Wuhan, China
    For correspondence
    jfliu@mail.hust.edu.cn
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0284-8377

Funding

Ministry of Science and Technology of the People's Republic of China (2018YFA0507003)

  • Jianfeng Liu

Agence Nationale de la Recherche (ANR-09-PIRI-0011)

  • Philippe Rondard

Fondation pour la recherche médicale FRM (FRM team: DEQ20170326522)

  • Jean-Philippe Pin

Spanish Ministry of Economy (SAF2015-74132-JIN)

  • Xavier Rovira

National Natural Science Foundation of China (81720108031)

  • Jianfeng Liu

National Natural Science Foundation of China (81872945)

  • Jianfeng Liu

National Natural Science Foundation of China (31721002)

  • Jianfeng Liu

National Natural Science Foundation of China (31420103909)

  • Jianfeng Liu

Ministry of Education of the People's Republic of China (B08029)

  • Jianfeng Liu

Centre National de la Recherche Scientifique (PICS n{degree sign}07030)

  • Philippe Rondard

Centre National de la Recherche Scientifique (PRC n{degree sign}1403)

  • Philippe Rondard

Institut National de la Santé et de la Recherche Médicale (IRP Brain Signal)

  • Philippe Rondard

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

© 2021, Liu et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 2,191
    views
  • 443
    downloads
  • 24
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Lei Liu
  2. Zhiran Fan
  3. Xavier Rovira
  4. Li Xue
  5. Salomé Roux
  6. Isabelle Brabet
  7. Mingxia Xin
  8. Jean-Philippe Pin
  9. Philippe Rondard
  10. Jianfeng Liu
(2021)
Allosteric ligands control the activation of a class C GPCR heterodimer by acting at the transmembrane interface
eLife 10:e70188.
https://doi.org/10.7554/eLife.70188

Share this article

https://doi.org/10.7554/eLife.70188

Further reading

    1. Biochemistry and Chemical Biology
    Nelson García-Vázquez, Tania J González-Robles ... Michele Pagano
    Research Article

    In healthy cells, cyclin D1 is expressed during the G1 phase of the cell cycle, where it activates CDK4 and CDK6. Its dysregulation is a well-established oncogenic driver in numerous human cancers. The cancer-related function of cyclin D1 has been primarily studied by focusing on the phosphorylation of the retinoblastoma (RB) gene product. Here, using an integrative approach combining bioinformatic analyses and biochemical experiments, we show that GTSE1 (G-Two and S phases expressed protein 1), a protein positively regulating cell cycle progression, is a previously unrecognized substrate of cyclin D1–CDK4/6 in tumor cells overexpressing cyclin D1 during G1 and subsequent phases. The phosphorylation of GTSE1 mediated by cyclin D1–CDK4/6 inhibits GTSE1 degradation, leading to high levels of GTSE1 across all cell cycle phases. Functionally, the phosphorylation of GTSE1 promotes cellular proliferation and is associated with poor prognosis within a pan-cancer cohort. Our findings provide insights into cyclin D1’s role in cell cycle control and oncogenesis beyond RB phosphorylation.

    1. Biochemistry and Chemical Biology
    2. Microbiology and Infectious Disease
    Mai Nguyen, Elda Bauda ... Cecile Morlot
    Research Article

    Teichoic acids (TA) are linear phospho-saccharidic polymers and important constituents of the cell envelope of Gram-positive bacteria, either bound to the peptidoglycan as wall teichoic acids (WTA) or to the membrane as lipoteichoic acids (LTA). The composition of TA varies greatly but the presence of both WTA and LTA is highly conserved, hinting at an underlying fundamental function that is distinct from their specific roles in diverse organisms. We report the observation of a periplasmic space in Streptococcus pneumoniae by cryo-electron microscopy of vitreous sections. The thickness and appearance of this region change upon deletion of genes involved in the attachment of TA, supporting their role in the maintenance of a periplasmic space in Gram-positive bacteria as a possible universal function. Consequences of these mutations were further examined by super-resolved microscopy, following metabolic labeling and fluorophore coupling by click chemistry. This novel labeling method also enabled in-gel analysis of cell fractions. With this approach, we were able to titrate the actual amount of TA per cell and to determine the ratio of WTA to LTA. In addition, we followed the change of TA length during growth phases, and discovered that a mutant devoid of LTA accumulates the membrane-bound polymerized TA precursor.