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

Author:

Hüsler Dario1,Stauffer Pia1ORCID,Keller Bernhard1,Böck Desirée2,Steiner Thomas3,Ostrzinski Anne4,Vormittag Simone1,Striednig Bianca1ORCID,Swart A Leoni1,Letourneur François5ORCID,Maaß Sandra4,Becher Dörte4ORCID,Eisenreich Wolfgang3,Pilhofer Martin2,Hilbi Hubert1ORCID

Affiliation:

1. Institute of Medical Microbiology, University of Zürich

2. Institute of Molecular Biology and Biophysics, ETH Zürich

3. Bavarian NMR Center - Structural Membrane Biochemistry, School of Natural Sciences, Technical University of Munich

4. Institute of Microbiology, University of Greifswald

5. UMR5294, LPHI, CNRS, INSERM, University of Montpellier

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-LD 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-LD interactions. In vitro reconstitution using purified LCVs and LDs from parental or Δsey1 mutant D. discoideum indicated that Sey1 and GTP promote this process. Sey1 and the L. pneumophila fatty acid transporter FadL were 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.

Funder

Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung

NOMIS Stiftung

Publisher

eLife Sciences Publications, Ltd

Subject

General Immunology and Microbiology,General Biochemistry, Genetics and Molecular Biology,General Medicine,General Neuroscience

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