Structure of Mycobacterium tuberculosis Cya, an evolutionary ancestor of the mammalian membrane adenylyl cyclases

  1. Ved Mehta
  2. Basavraj Khanppnavar
  3. Dina Schuster
  4. Ilayda Kantarci
  5. Irene Vercellino
  6. Angela Kosturanova
  7. Tarun Iype
  8. Sasa Stefanic
  9. Paola Picotti
  10. Volodymyr M. Korkhov  Is a corresponding author
  1. Paul Scherrer Institute, Switzerland
  2. ETH Zurich, Switzerland
  3. University of Zurich, Switzerland

Abstract

Mycobacterium tuberculosis adenylyl cyclase (AC) Rv1625c / Cya is an evolutionary ancestor of the mammalian membrane ACs and a model system for studies of their structure and function. Although the vital role of ACs in cellular signaling is well established, the function of their transmembrane (TM) regions remains unknown. Here we describe the cryo-EM structure of Cya bound to a stabilizing nanobody at 3.6 Å resolution. The TM helices 1-5 form a structurally conserved domain that facilitates the assembly of the helical and catalytic domains. The TM region contains discrete pockets accessible from the extracellular and cytosolic side of the membrane. Neutralization of the negatively charged extracellular pocket Ex1 destabilizes the cytosolic helical domain and reduces the catalytic activity of the enzyme. The TM domain acts as a functional component of Cya, guiding the assembly of the catalytic domain and providing the means for direct regulation of catalytic activity in response to extracellular ligands.

Data availability

The atomic coordinates and structure factors have been deposited in the Protein Data Bank (7YZ9, 7YZI, 7YZK); the density maps have been deposited in the Electron Microscopy Data Bank (EMD-14388, EMD-14389). The mass spectrometry data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD033826. All other data are available in the main text or the supplementary materials.

The following data sets were generated

Article and author information

Author details

  1. Ved Mehta

    Laboratory of Biomolecular Research, Paul Scherrer Institute, Villigen, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  2. Basavraj Khanppnavar

    Laboratory of Biomolecular Research, Paul Scherrer Institute, Villigen, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  3. Dina Schuster

    Institute of Molecular Biology and Biophysics, ETH Zurich, Zurich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6611-8237
  4. Ilayda Kantarci

    Laboratory of Biomolecular Research, Paul Scherrer Institute, Villigen, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  5. Irene Vercellino

    Laboratory of Biomolecular Research, Paul Scherrer Institute, Villigen, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  6. Angela Kosturanova

    Laboratory of Biomolecular Research, Paul Scherrer Institute, Villigen, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  7. Tarun Iype

    Laboratory of Biomolecular Research, Paul Scherrer Institute, Villigen, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  8. Sasa Stefanic

    Institute of Parasitology, University of Zurich, Zurich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7367-1831
  9. Paola Picotti

    Institute of Molecular Systems Biology, ETH Zurich, Zurich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  10. Volodymyr M. Korkhov

    Laboratory of Biomolecular Research, Paul Scherrer Institute, Villigen, Switzerland
    For correspondence
    volodymyr.korkhov@psi.ch
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0962-9433

Funding

Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (150665)

  • Volodymyr M. Korkhov

Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (176992)

  • Volodymyr M. Korkhov

Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (184951)

  • Volodymyr M. Korkhov

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

Reviewing Editor

  1. Andrew P Carter, MRC Laboratory of Molecular Biology, United Kingdom

Publication history

  1. Preprint posted: December 1, 2021 (view preprint)
  2. Received: January 13, 2022
  3. Accepted: August 17, 2022
  4. Accepted Manuscript published: August 18, 2022 (version 1)
  5. Accepted Manuscript updated: August 22, 2022 (version 2)
  6. Version of Record published: August 31, 2022 (version 3)

Copyright

© 2022, Mehta 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.

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  1. Ved Mehta
  2. Basavraj Khanppnavar
  3. Dina Schuster
  4. Ilayda Kantarci
  5. Irene Vercellino
  6. Angela Kosturanova
  7. Tarun Iype
  8. Sasa Stefanic
  9. Paola Picotti
  10. Volodymyr M. Korkhov
(2022)
Structure of Mycobacterium tuberculosis Cya, an evolutionary ancestor of the mammalian membrane adenylyl cyclases
eLife 11:e77032.
https://doi.org/10.7554/eLife.77032
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