The large GTPase Sey1/atlastin mediates lipid droplet- and FadL-dependent intracellular fatty acid metabolism of Legionella pneumophila
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...
| Main Authors: | , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2023-05-01
|
| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/85142 |
| _version_ | 1797810045657808896 |
|---|---|
| author | 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 |
| author_facet | 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 |
| author_sort | Dario Hüsler |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-13T07:02:12Z |
| format | Article |
| id | doaj.art-657f53154fbe40e1be705fd5ac2662de |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-03-13T07:02:12Z |
| publishDate | 2023-05-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-657f53154fbe40e1be705fd5ac2662de2023-06-06T16:16:20ZengeLife Sciences Publications LtdeLife2050-084X2023-05-011210.7554/eLife.85142The large GTPase Sey1/atlastin mediates lipid droplet- and FadL-dependent intracellular fatty acid metabolism of Legionella pneumophilaDario Hüsler0Pia Stauffer1https://orcid.org/0009-0008-5742-8362Bernhard Keller2Desirée Böck3Thomas Steiner4Anne Ostrzinski5Simone Vormittag6Bianca Striednig7https://orcid.org/0000-0001-7575-8965A Leoni Swart8François Letourneur9https://orcid.org/0000-0003-2232-6127Sandra Maaß10Dörte Becher11https://orcid.org/0000-0002-9630-5735Wolfgang Eisenreich12Martin Pilhofer13Hubert Hilbi14https://orcid.org/0000-0002-5462-9301Institute of Medical Microbiology, University of Zürich, Zürich, SwitzerlandInstitute of Medical Microbiology, University of Zürich, Zürich, SwitzerlandInstitute of Medical Microbiology, University of Zürich, Zürich, SwitzerlandInstitute of Molecular Biology and Biophysics, ETH Zürich, Zürich, SwitzerlandBavarian NMR Center - Structural Membrane Biochemistry, School of Natural Sciences, Technical University of Munich, Garching, GermanyInstitute of Microbiology, University of Greifswald, Greifswald, GermanyInstitute of Medical Microbiology, University of Zürich, Zürich, SwitzerlandInstitute of Medical Microbiology, University of Zürich, Zürich, SwitzerlandInstitute of Medical Microbiology, University of Zürich, Zürich, SwitzerlandUMR5294, LPHI, CNRS, INSERM, University of Montpellier, Montpellier, FranceInstitute of Microbiology, University of Greifswald, Greifswald, GermanyInstitute of Microbiology, University of Greifswald, Greifswald, GermanyBavarian NMR Center - Structural Membrane Biochemistry, School of Natural Sciences, Technical University of Munich, Garching, GermanyInstitute of Molecular Biology and Biophysics, ETH Zürich, Zürich, SwitzerlandInstitute of Medical Microbiology, University of Zürich, Zürich, SwitzerlandThe 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.https://elifesciences.org/articles/85142host-pathogen interactionintracellular bacterialarge GTPaseLegionellalipid dropletpathogen vacuole |
| spellingShingle | 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 The large GTPase Sey1/atlastin mediates lipid droplet- and FadL-dependent intracellular fatty acid metabolism of Legionella pneumophila eLife host-pathogen interaction intracellular bacteria large GTPase Legionella lipid droplet pathogen vacuole |
| title | The large GTPase Sey1/atlastin mediates lipid droplet- and FadL-dependent intracellular fatty acid metabolism of Legionella pneumophila |
| title_full | The large GTPase Sey1/atlastin mediates lipid droplet- and FadL-dependent intracellular fatty acid metabolism of Legionella pneumophila |
| title_fullStr | The large GTPase Sey1/atlastin mediates lipid droplet- and FadL-dependent intracellular fatty acid metabolism of Legionella pneumophila |
| title_full_unstemmed | The large GTPase Sey1/atlastin mediates lipid droplet- and FadL-dependent intracellular fatty acid metabolism of Legionella pneumophila |
| title_short | The large GTPase Sey1/atlastin mediates lipid droplet- and FadL-dependent intracellular fatty acid metabolism of Legionella pneumophila |
| title_sort | large gtpase sey1 atlastin mediates lipid droplet and fadl dependent intracellular fatty acid metabolism of legionella pneumophila |
| topic | host-pathogen interaction intracellular bacteria large GTPase Legionella lipid droplet pathogen vacuole |
| url | https://elifesciences.org/articles/85142 |
| work_keys_str_mv | AT dariohusler thelargegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT piastauffer thelargegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT bernhardkeller thelargegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT desireebock thelargegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT thomassteiner thelargegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT anneostrzinski thelargegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT simonevormittag thelargegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT biancastriednig thelargegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT aleoniswart thelargegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT francoisletourneur thelargegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT sandramaaß thelargegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT dortebecher thelargegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT wolfgangeisenreich thelargegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT martinpilhofer thelargegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT huberthilbi thelargegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT dariohusler largegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT piastauffer largegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT bernhardkeller largegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT desireebock largegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT thomassteiner largegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT anneostrzinski largegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT simonevormittag largegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT biancastriednig largegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT aleoniswart largegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT francoisletourneur largegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT sandramaaß largegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT dortebecher largegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT wolfgangeisenreich largegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT martinpilhofer largegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila AT huberthilbi largegtpasesey1atlastinmediateslipiddropletandfadldependentintracellularfattyacidmetabolismoflegionellapneumophila |