Nanobodies combined with DNA-PAINT super-resolution reveal a staggered titin nano-architecture in flight muscles

  1. Florian Schueder
  2. Pierre Mangeol
  3. Eunice HoYee Chan
  4. Renate Rees
  5. Jürgen Schünemann
  6. Ralf Jungmann  Is a corresponding author
  7. Dirk Görlich  Is a corresponding author
  8. Frank Schnorrer  Is a corresponding author
  1. Max Planck Institute of Biochemistry, Germany
  2. Aix Marseille University, CNRS, IDBM, France
  3. Max Planck Institute for Multidisciplinary Sciences, Germany
  4. Ludwig Maximilian University, Germany

Abstract

Sarcomeres are the force producing units of all striated muscles. Their nanoarchitecture critically depends on the large titin protein, which in vertebrates spans from the sarcomeric Z-disc to the M-band and hence links actin and myosin filaments stably together. This ensures sarcomeric integrity and determines the length of vertebrate sarcomeres. However, the instructive role of titins for sarcomeric architecture outside of vertebrates is not as well understood. Here, we used a series of nanobodies, the Drosophila titin nanobody toolbox, recognising specific domains of the two Drosophila titin homologs Sallimus and Projectin to determine their precise location in intact flight muscles. By combining nanobodies with DNA-PAINT super-resolution microscopy, we found that, similar to vertebrate titin, Sallimus bridges across the flight muscle I-band, whereas Projectin is located at the beginning of the A-band. Interestingly, the ends of both proteins overlap at the I-band/A-band border, revealing a staggered organisation of the two Drosophila titin homologs. This architecture may help to stably anchor Sallimus at the myosin filament and hence ensure efficient force transduction during flight.

Data availability

All source data for all figures are provided.Figure 3 - Source Data 1Figure 4 - Source Data 1Figure 5 - Source Data 1Figure 6 - Source Data 1All new code has been uploaded to a public database and the link is provided.

The following data sets were generated
    1. Pierre Mangeol
    (2022) titin_PAINT
    Github, PierreMangeol/titin_PAINT.

Article and author information

Author details

  1. Florian Schueder

    Max Planck Institute of Biochemistry, Martinsried, Germany
    Competing interests
    The authors declare that no competing interests exist.
  2. Pierre Mangeol

    Turing Centre for Living Systems, Aix Marseille University, CNRS, IDBM, Marseille, France
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8305-7322
  3. Eunice HoYee Chan

    Turing Centre for Living Systems, Aix Marseille University, CNRS, IDBM, Marseille, France
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3162-3609
  4. Renate Rees

    Department of Cellular Logistics, Max Planck Institute for Multidisciplinary Sciences, Göttingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  5. Jürgen Schünemann

    Department of Cellular Logistics, Max Planck Institute for Multidisciplinary Sciences, Göttingen, Germany
    Competing interests
    The authors declare that no competing interests exist.
  6. Ralf Jungmann

    Ludwig Maximilian University, Munich, Germany
    For correspondence
    jungmann@biochem.mpg.de
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4607-3312
  7. Dirk Görlich

    Department of Cellular Logistics, Max Planck Institute for Multidisciplinary Sciences, Göttingen, Germany
    For correspondence
    goerlich@mpinat.mpg.de
    Competing interests
    The authors declare that no competing interests exist.
  8. Frank Schnorrer

    Turing Centre for Living Systems, Aix Marseille University, CNRS, IDBM, Marseille, France
    For correspondence
    frank.schnorrer@univ-amu.fr
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9518-7263

Funding

Centre National de la Recherche Scientifique

  • Frank Schnorrer

Human Frontier Science Program (RGP0052/2018)

  • Frank Schnorrer

Agence Nationale de la Recherche (ANR-10-INBS-04-01)

  • Frank Schnorrer

Agence Nationale de la Recherche (ANR-16-CONV-0001,Turing Centre for Living Systems)

  • Frank Schnorrer

Aix-Marseille Université

  • Pierre Mangeol

Max-Planck-Gesellschaft

  • Dirk Görlich

Max-Planck-Gesellschaft

  • Ralf Jungmann

European Research Council (ERC-2019-SyG 856118)

  • Dirk Görlich

European Research Council (ERC-2019-SyG 856118)

  • Frank Schnorrer

European Research Council (ERC-2015-StG 680241)

  • Ralf Jungmann

Aix-Marseille Université (ANR-11-IDEX-0001-02)

  • Frank Schnorrer

Agence Nationale de la Recherche (ACHN MUSCLE-FORCES)

  • Frank Schnorrer

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

Reviewing Editor

  1. Michel Labouesse, UMR7622, Institut de Biologie Paris-Seine, Sorbonne Université, France

Publication history

  1. Received: April 7, 2022
  2. Preprint posted: April 15, 2022 (view preprint)
  3. Accepted: November 22, 2022
  4. Accepted Manuscript published: January 16, 2023 (version 1)

Copyright

© 2023, Schueder 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. Florian Schueder
  2. Pierre Mangeol
  3. Eunice HoYee Chan
  4. Renate Rees
  5. Jürgen Schünemann
  6. Ralf Jungmann
  7. Dirk Görlich
  8. Frank Schnorrer
(2023)
Nanobodies combined with DNA-PAINT super-resolution reveal a staggered titin nano-architecture in flight muscles
eLife 12:e79344.
https://doi.org/10.7554/eLife.79344

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