Porcellato, Elena and Gonzalez-Sanchez, Juan Carlos and Ahlmann-Eltze, Constantin and Elsakka, Mahmoud Ali and Shapira, Itamar and Fritsch, Juergen and Navarro, Juan Antonio and Anders, Simon and Russell, Robert B. and Wieland, Felix T. and Metzendorf, Christoph (2022) The S-palmitoylome and DHHC-PAT interactome of Drosophila melanogaster S2R+cells indicate a high degree of conservation to mammalian palmitoylomes. PLOS ONE, 17 (8). ISSN 1932-6203,
Full text not available from this repository. (Request a copy)Abstract
Protein S-palmitoylation, the addition of a long-chain fatty acid to target proteins, is among the most frequent reversible protein modifications in Metazoa, affecting subcellular protein localization, trafficking and protein-protein interactions. S-palmitoylated proteins are abundant in the neuronal system and are associated with neuronal diseases and cancer. Despite the importance of this post-translational modification, it has not been thoroughly studied in the model organism Drosophila melanogaster. Here we present the palmitoylome of Drosophila S2R+ cells, comprising 198 proteins, an estimated 3.5% of expressed genes in these cells. Comparison of orthologs between mammals and Drosophila suggests that S-palmitoylated proteins are more conserved between these distant phyla than non-S-palmitoylated proteins. To identify putative client proteins and interaction partners of the DHHC family of protein acyl-transferases (PATs) we established DHHC-BioID, a proximity biotinylation-based method. In S2R+ cells, ectopic expression of the DHHC-PAT dHip14-BioID in combination with Snap24 or an interaction-deficient Snap24-mutant as a negative control, resulted in biotinylation of Snap24 but not the Snap24-mutant. DHHC-BioID in S2R+ cells using 10 different DHHC-PATs as bait identified 520 putative DHHC-PAT interaction partners of which 48 were S-palmitoylated and are therefore putative DHHC-PAT client proteins. Comparison of putative client protein/DHHC-PAT combinations indicates that CG8314, CG5196, CG5880 and Patsas have a preference for transmembrane proteins, while S-palmitoylated proteins with the Hip14-interaction motif are most enriched by DHHC-BioID variants of approximated and dHip14. Finally, we show that BioID is active in larval and adult Drosophila and that dHip14-BioID rescues dHip14 mutant flies, indicating that DHHC-BioID is non-toxic. In summary we provide the first systematic analysis of a Drosophila palmitoylome. We show that DHHC-BioID is sensitive and specific enough to identify DHHC-PAT client proteins and provide DHHC-PAT assignment for ca. 25% of the S2R+ cell palmitoylome, providing a valuable resource. In addition, we establish DHHC-BioID as a useful concept for the identification of tissue-specific DHHC-PAT interactomes in Drosophila.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | PROTEIN PALMITOYLATION; ZDHHC FAMILY; EXPRESSION; IDENTIFICATION; RECEPTOR; LOCALIZATION; THIOESTERASE; INHIBITION; ACYLATION; REQUIRES; |
| Subjects: | 500 Science > 590 Zoological sciences |
| Divisions: | Medicine > Abteilung für Krankenhaushygiene und Infektiologie Biology, Preclinical Medicine > Institut für Zoologie > Entwicklungsbiologie (Prof. Dr. Stephan Schneuwly) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 20 Feb 2024 16:12 |
| Last Modified: | 20 Feb 2024 16:12 |
| URI: | https://pred.uni-regensburg.de/id/eprint/57737 |
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