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Kuhn L., Majzoub K., Einhorn E., Chicher J., Pompon Julien, Imler J. L., Hammann P., Meignin C. (2017). Definition of a RACK1 interaction network in Drosophila melanogaster using SWATH-MS. G3 : Genes Genomes Genetics, 7 (7), 2249-2258. ISSN 2160-1836

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Lien direct chez l'éditeur doi:10.1534/g3.117.042564

Titre
Definition of a RACK1 interaction network in Drosophila melanogaster using SWATH-MS
Année de publication2017
Type de documentArticle référencé dans le Web of Science WOS:000404991600022
AuteursKuhn L., Majzoub K., Einhorn E., Chicher J., Pompon Julien, Imler J. L., Hammann P., Meignin C.
SourceG3 : Genes Genomes Genetics, 2017, 7 (7), p. 2249-2258. ISSN 2160-1836
RésuméReceptor for Activated protein C kinase 1 (RACK1) is a scaffold protein that has been found in association with several signaling complexes, and with the 40S subunit of the ribosome. Using the model organism Drosophila melanogaster, we recently showed that RACK1 is required at the ribosome for internal ribosome entry site (IRES)-mediated translation of viruses. Here, we report a proteomic characterization of the interactome of RACK1 in Drosophila S2 cells. We carried out Label-Free quantitation using both Data-Dependent and Data-Independent Acquisition (DDA and DIA, respectively) and observed a significant advantage for the Sequential Window Acquisition of all THeoretical fragment-ion spectra (SWATH) method, both in terms of identification of interactants and quantification of low abundance proteins. These data represent the first SWATH spectral library available for Drosophila and will be a useful resource for the community. A total of 52 interacting proteins were identified, including several molecules involved in translation such as structural components of the ribosome, factors regulating translation initiation or elongation, and RNA binding proteins. Among these 52 proteins, 15 were identified as partners by the SWATH strategy only. Interestingly, these 15 proteins are significantly enriched for the functions translation and nucleic acid binding. This enrichment reflects the engagement of RACK1 at the ribosome and highlights the added value of SWATH analysis. A functional screen did not reveal any protein sharing the interesting properties of RACK1, which is required for IRES-dependent translation and not essential for cell viability. Intriguingly however, 10 of the RACK1 partners identified restrict replication of Cricket paralysis virus (CrPV), an IRES-containing virus.
Plan de classementSciences fondamentales / Techniques d'analyse et de recherche [020] ; Entomologie médicale / Parasitologie / Virologie [052]
LocalisationFonds IRD [F B010070331]
Identifiant IRDfdi:010070331
Lien permanenthttp://www.documentation.ird.fr/hor/fdi:010070331

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