The S-palmitoylome and DHHC-PAT interactome of Drosophila melanogaster S2R+ cells indicate a high degree of conservation to mammalian palmitoylomes

• Verfasst von: Porcellato, Elena [VerfasserIn]
• Verfasst von: González Sánchez, Juan Carlos [VerfasserIn]
• Verfasst von: Ahlmann-Eltze, Constantin [VerfasserIn]
• Verfasst von: Elsakka, Mahmoud Ali [VerfasserIn]
• Verfasst von: Shapira, Itamar [VerfasserIn]
• Verfasst von: Fritsch, Jürgen [VerfasserIn]
• Verfasst von: Navarro, Juan Antonio [VerfasserIn]
• Verfasst von: Anders, Simon [VerfasserIn]
• Verfasst von: Russell, Robert B. [VerfasserIn]
• Verfasst von: Wieland, Felix T. [VerfasserIn]
• Verfasst von: Metzendorf, Christoph [VerfasserIn]
• Titel: The S-palmitoylome and DHHC-PAT interactome of Drosophila melanogaster S2R+ cells indicate a high degree of conservation to mammalian palmitoylomes
• Verf.angabe: Elena Porcellato, Juan Carlos González-Sánchez, Constantin Ahlmann-Eltze, Mahmoud Ali Elsakka, Itamar Shapira, Jürgen Fritsch, Juan Antonio Navarro, Simon Anders, Robert B. Russell, Felix T. Wieland, Christoph Metzendorf
• E-Jahr: 2022
• Jahr: August 12, 2022
• Umfang: 24 S.
• Fussnoten: Gesehen am 04.04.2023
• Titel Quelle: Enthalten in: PLOS ONE
• Ort Quelle: San Francisco, California, US : PLOS, 2006
• Jahr Quelle: 2022
• Band/Heft Quelle: 17(2022), 8 vom: Aug., Artikel-ID e0261543, Seite 1-24
• ISSN Quelle: 1932-6203
• DOI: doi:10.1371/journal.pone.0261543
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: Biotinylation
• Sach-SW: Chaperone proteins
• Sach-SW: Drosophila melanogaster
• Sach-SW: Larvae
• Sach-SW: Mammals
• Sach-SW: Membrane proteins
• Sach-SW: Palmitoylation
• Sach-SW: Protein interactions
• K10plus-PPN: 1841176923

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.