eLife (May 2023)

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

  • Dario Hüsler,
  • Pia Stauffer,
  • Bernhard Keller,
  • Desirée Böck,
  • Thomas Steiner,
  • Anne Ostrzinski,
  • Simone Vormittag,
  • Bianca Striednig,
  • A Leoni Swart,
  • François Letourneur,
  • Sandra Maaß,
  • Dörte Becher,
  • Wolfgang Eisenreich,
  • Martin Pilhofer,
  • Hubert Hilbi

DOI
https://doi.org/10.7554/eLife.85142
Journal volume & issue
Vol. 12

Abstract

Read online

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.

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