The Large GTPase Sey1/atlastin Mediates Lipid Droplet- and FadL-dependent Intracellular Fatty Acid Metabolism of

Overview

Journal Elife

Specialty Biology

Date 2023 May 9

PMID 37158597

Authors

Dario Husler

Pia Stauffer

Bernhard Keller

Desiree Bock

Thomas Steiner

Anne Ostrzinski

Simone Vormittag

Bianca Striednig

A Leoni Swart

Francois Letourneur

Sandra Maass

Dorte Becher

Wolfgang Eisenreich

Martin Pilhofer

Hubert Hilbi

Affiliations

Soon will be listed here.

Abstract

The amoeba-resistant bacterium causes Legionnaires' disease and employs a type IV secretion system (T4SS) to replicate in the unique, ER-associated -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 . Dually fluorescence-labeled producing LCV and LD markers revealed that Sey1 as well as the T4SS and the Ran GTPase activator LegG1 promote LCV-LD interactions. In vitro reconstitution using purified LCVs and LDs from parental or Δ mutant indicated that Sey1 and GTP promote this process. Sey1 and the 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 .

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References

Hu J, Rapoport T . Fusion of the endoplasmic reticulum by membrane-bound GTPases. Semin Cell Dev Biol. 2016; 60:105-111. DOI: 10.1016/j.semcdb.2016.06.001. View

Du X, Barisch C, Paschke P, Herrfurth C, Bertinetti O, Pawolleck N . Dictyostelium lipid droplets host novel proteins. Eukaryot Cell. 2013; 12(11):1517-29. PMC: 3837934. DOI: 10.1128/EC.00182-13. View

Sherwood R, Roy C . Autophagy Evasion and Endoplasmic Reticulum Subversion: The Yin and Yang of Legionella Intracellular Infection. Annu Rev Microbiol. 2016; 70:413-33. DOI: 10.1146/annurev-micro-102215-095557. View

Voeltz G, Prinz W, Shibata Y, Rist J, Rapoport T . A class of membrane proteins shaping the tubular endoplasmic reticulum. Cell. 2006; 124(3):573-86. DOI: 10.1016/j.cell.2005.11.047. View

Finsel I, Ragaz C, Hoffmann C, Harrison C, Weber S, van Rahden V . The Legionella effector RidL inhibits retrograde trafficking to promote intracellular replication. Cell Host Microbe. 2013; 14(1):38-50. DOI: 10.1016/j.chom.2013.06.001. View

Weber S, Hilbi H . Live cell imaging of phosphoinositide dynamics during Legionella infection. Methods Mol Biol. 2014; 1197:153-67. DOI: 10.1007/978-1-4939-1261-2_9. View

Byrnes L, Singh A, Szeto K, Benvin N, ODonnell J, Zipfel W . Structural basis for conformational switching and GTP loading of the large G protein atlastin. EMBO J. 2013; 32(3):369-84. PMC: 3567502. DOI: 10.1038/emboj.2012.353. View

Recuero-Checa M, Sharma M, Lau C, Watkins P, Gaydos C, Dean D . Chlamydia trachomatis growth and development requires the activity of host Long-chain Acyl-CoA Synthetases (ACSLs). Sci Rep. 2016; 6:23148. PMC: 4796813. DOI: 10.1038/srep23148. View

Marko M, Hsieh C, Schalek R, Frank J, Mannella C . Focused-ion-beam thinning of frozen-hydrated biological specimens for cryo-electron microscopy. Nat Methods. 2007; 4(3):215-7. DOI: 10.1038/nmeth1014. View

10.

Manske C, Schell U, Hilbi H . Metabolism of myo-Inositol by Legionella pneumophila Promotes Infection of Amoebae and Macrophages. Appl Environ Microbiol. 2016; 82(16):5000-14. PMC: 4968532. DOI: 10.1128/AEM.01018-16. View

11.

Anwar K, Klemm R, Condon A, Severin K, Zhang M, Ghirlando R . The dynamin-like GTPase Sey1p mediates homotypic ER fusion in S. cerevisiae. J Cell Biol. 2012; 197(2):209-17. PMC: 3328390. DOI: 10.1083/jcb.201111115. View

12.

Swart A, Gomez-Valero L, Buchrieser C, Hilbi H . Evolution and function of bacterial RCC1 repeat effectors. Cell Microbiol. 2020; 22(10):e13246. DOI: 10.1111/cmi.13246. View

13.

Barisch C, Soldati T . Breaking fat! How mycobacteria and other intracellular pathogens manipulate host lipid droplets. Biochimie. 2017; 141:54-61. DOI: 10.1016/j.biochi.2017.06.001. View

14.

Bosch M, Sweet M, Parton R, Pol A . Lipid droplets and the host-pathogen dynamic: FATal attraction?. J Cell Biol. 2021; 220(8). PMC: 8240858. DOI: 10.1083/jcb.202104005. View

15.

Schell U, Simon S, Sahr T, Hager D, Albers M, Kessler A . The α-hydroxyketone LAI-1 regulates motility, Lqs-dependent phosphorylation signalling and gene expression of Legionella pneumophila. Mol Microbiol. 2015; 99(4):778-93. DOI: 10.1111/mmi.13265. View

16.

Wilfling F, Thiam A, Olarte M, Wang J, Beck R, Gould T . Arf1/COPI machinery acts directly on lipid droplets and enables their connection to the ER for protein targeting. Elife. 2014; 3:e01607. PMC: 3913038. DOI: 10.7554/eLife.01607. View

17.

Nixon-Abell J, Obara C, Weigel A, Li D, Legant W, Xu C . Increased spatiotemporal resolution reveals highly dynamic dense tubular matrices in the peripheral ER. Science. 2016; 354(6311). PMC: 6528812. DOI: 10.1126/science.aaf3928. View

18.

Kory N, Farese Jr R, Walther T . Targeting Fat: Mechanisms of Protein Localization to Lipid Droplets. Trends Cell Biol. 2016; 26(7):535-546. PMC: 4976449. DOI: 10.1016/j.tcb.2016.02.007. View

19.

Bender J, Rydzewski K, Broich M, Schunder E, Heuner K, Flieger A . Phospholipase PlaB of Legionella pneumophila represents a novel lipase family: protein residues essential for lipolytic activity, substrate specificity, and hemolysis. J Biol Chem. 2009; 284(40):27185-94. PMC: 2785646. DOI: 10.1074/jbc.M109.026021. View

20.

Hackstadt T, Rockey D, Heinzen R, Scidmore M . Chlamydia trachomatis interrupts an exocytic pathway to acquire endogenously synthesized sphingomyelin in transit from the Golgi apparatus to the plasma membrane. EMBO J. 1996; 15(5):964-77. PMC: 449991. View