PXD038200

PXD038200 is an original dataset announced via ProteomeXchange.

Title	The large GTPase Sey1/atlastin mediates lipid droplet- and FadL-dependent intracellular fatty acid metabolism of Legionella pneumophila
Description	The facultative intracellular bacterium Legionella pneumophila employs the Icm/Dot type IV secretion system (T4SS) to replicate in a unique membrane-bound compartment, the Legionella-containing vacuole (LCV). The ER-resident large fusion GTPase Sey1/atlastin-3 promotes remodeling and expansion of LCVs, and the GTPase is also implicated in the formation of ER-derived lipid droplets (LDs). Here we reveal that LCVs intimately interact with palmitate-induced LDs in Dictyostelium discoideum amoeba. Comparative proteomics of LDs isolated from the D. discoideum parental strain Ax3 or sey1 revealed 192 differentially produced proteins, of which 7 or 22 were exclusively detected in LDs isolated from strain Ax3 or sey1, respectively. Using dually fluorescence-labeled amoeba producing the LCV marker P4C-GFP or AmtA-GFP and the LD marker mCherry-perilipin, we show that Sey1 and the L. pneumophila Icm/Dot T4SS as well as the effector LegG1 promote LCV-LD interactions. In vitro reconstitution of the LCV-LD interactions using purified LCVs and LDs from D. discoideum Ax3 or sey1 revealed that Sey1 and GTP promote this process. The LCV-LD interactions were impaired forsey1-derived LDs, suggesting that Sey1 regulates LD traits. Palmitate promoted the growth of L. pneumophila in D. discoideum Ax3 but not in sey1 and of wild-type but not mutant bacteria lacking a homologue of the E. coli fatty acid transporter FadL. Finally, isotopologue profiling indicated that intracellular L. pneumophila metabolizes 13C-palmitate, and its catabolism was reduced in D. discoideum sey1 and L. pneumophila fadL. Taken together, our results reveal that Sey1 mediates LD- and FadL-dependent fatty acid metabolism of intracellular L. pneumophila
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_08:59:49.206.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Sandra Maass
SpeciesList	scientific name: Dictyostelium discoideum (Slime mold); NCBI TaxID: 44689;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2022-11-17 07:55:00	ID requested
1	2023-06-06 23:04:22	announced
⏵ 2	2023-11-14 08:59:49	announced	2023-11-14: Updated project metadata.

Dataset with its publication pending

submitter keyword: large fusion GTPase, Legionella pneumophila, atlastin, lipid droplet, Legionnaires’ disease, host-pathogen interaction, pathogen vacuole, fatty acid transport,Amoeba, Dictyostelium discoideum

Dörte Becher
contact affiliation	Institute of Microbiology, University of Greifswald, Felix-Hausdorff-Strasse 8, 17489 Greifswald, Germany
contact email	dbecher@uni-greifswald.de
lab head
Sandra Maass
contact affiliation	University of Greifswald, Department for Microbial Proteomics
contact email	sandra.maass@uni-greifswald.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD038200
     1. Label: PRIDE project
     2. Name: The large GTPase Sey1/atlastin mediates lipid droplet- and FadL-dependent intracellular fatty acid metabolism of Legionella pneumophila