The large GTPase Sey1/atlastin mediates lipid droplet- and FadL-dependent intracellular fatty acid metabolism of Legionella pneumophila

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Abstract

The amoeba-resistant bacterium Legionella pneumophila causes Legionnaires' disease and employs a type IV secretion system (T4SS) to replicate in the unique, ER-associated Legionella-containing vacuole (LCV). The large fusion GTPase Sey1/atlastin is implicated in ER dynamics, ER-derived lipid droplet (LD) formation, and LCV maturation. Here we employ cryo-electron tomography, confocal microscopy, proteomics, and isotopologue profiling to analyze LCV-LDs interactions in the genetically tractable amoeba Dictyostelium discoideum. Dually fluorescence-labeled D. discoideum producing LCV and LD markers revealed that Sey1 as well as the L. pneumophila T4SS and the Ran GTPase activator LegG1 promote LCV-LDs interactions. In vitro reconstitution using purified LCVs and LDs from parental or Dsey1 mutant D. discoideum indicated that Sey1 and GTP promote this process. Sey1 and the L. pneumophila fatty acid transporter FadL are implicated in palmitate catabolism and palmitate-dependent intracellular growth. Taken together, our results reveal that Sey1 and LegG1 mediate LD- and FadL-dependent fatty acid metabolism of intracellular L. pneumophila.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files.The MS proteomics data discussed in this publication have been deposited to the ProteomeXchange Consortium via the PRIDE (Perez-Riverol et al., 2019) partner repository with the dataset identifier PXD038200

The following data sets were generated

1. Hüsler D
2. Stauffer P
3. Keller B
4. Böck D
5. Steiner T
6. Ostrzinski A
7. Striednig B
8. Swart AL
9. Letourneur F
10. Maaß S
11. Becher D
12. Eisenreich W
13. Pilhofer M
14. Hilbi H
(2022) The large GTPase Sey1/atlastin mediates lipid droplet- and FadL-dependent intracellular fatty acid metabolism of Legionella pneumophila
PRIDE, PXD038200.

https://www.ebi.ac.uk/pride/archive/projects/PXD038200

Article and author information

Author details

Dario Hüsler

Institute of Medical Microbiology, University of Zurich, Zürich, Switzerland

Competing interests
The authors declare that no competing interests exist.
Pia Stauffer

Institute of Medical Microbiology, University of Zurich, Zürich, Switzerland

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The authors declare that no competing interests exist.
Bernhard Keller

Institute of Medical Microbiology, University of Zurich, Zürich, Switzerland

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The authors declare that no competing interests exist.
Desirée Böck

Institute of Molecular Biology and Biophysics, ETH Zurich, Zürich, Switzerland

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The authors declare that no competing interests exist.
Thomas Steiner

Bavarian NMR Center - Structural Membrane Biochemistry, Technical University of Munich, Garching, Germany

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The authors declare that no competing interests exist.
Anne Ostrzinski

Institute of Microbiology, University of Greifswald, Greifswald, Germany

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The authors declare that no competing interests exist.
Simone Vormittag

Institute of Medical Microbiology, University of Zurich, Zürich, Switzerland

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The authors declare that no competing interests exist.
Bianca Striednig

Institute of Medical Microbiology, University of Zurich, Zürich, Switzerland

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The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0001-7575-8965
A Leoni Swart

Institute of Medical Microbiology, University of Zurich, Zürich, Switzerland

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The authors declare that no competing interests exist.
François Letourneur

INSERM, University of Montpellier, Montpellier, France

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The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0003-2232-6127
Sandra Maaß

Institute of Microbiology, University of Greifswald, Greifswald, Germany

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The authors declare that no competing interests exist.
Dörte Becher

Institute for Microbiology, University of Greifswald, Greifswald, Germany

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The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-9630-5735
Wolfgang Eisenreich

Bavarian NMR Center - Structural Membrane Biochemistry, Technical University of Munich, Garching, Germany

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The authors declare that no competing interests exist.
Martin Pilhofer

Institute of Molecular Biology and Biophysics, ETH Zurich, Zürich, Switzerland

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The authors declare that no competing interests exist.
Hubert Hilbi

Institute of Medical Microbiology, University of Zurich, Zürich, Switzerland

For correspondence
hilbi@imm.uzh.ch

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The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-5462-9301

Funding

Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (31003A_175557)

Hubert Hilbi

Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (310030_207826)

Hubert Hilbi

NOMIS Stiftung

Martin Pilhofer

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

Copyright

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.