Please use this identifier to cite or link to this item: https://doi.org/10.1534/g3.117.042564
Title: Definition of a RACK1 interaction network in Drosophila melanogaster using SWATH-MS
Authors: Kuhn, L
Majzoub, K
Einhorn, E
Chicher, J
Pompon, J 
Imler, J.-L
Hammann, P
Meignin, C
Keywords: nucleic acid
receptor for activated C kinase 1
RNA binding protein
Drosophila protein
RACK1 protein, Drosophila
receptor for activated C kinase
viral protein
Article
cell viability
controlled study
Drosophila melanogaster
molecule
nonhuman
protein interaction
proteomics
ribosome
translation initiation
animal
biological model
biosynthesis
cell line
Dicistroviridae
Drosophila melanogaster
gene regulatory network
genetics
internal ribosome entry site
metabolism
protein synthesis
Animals
Cell Line
Dicistroviridae
Drosophila melanogaster
Drosophila Proteins
Gene Regulatory Networks
Internal Ribosome Entry Sites
Models, Genetic
Protein Biosynthesis
Receptors for Activated C Kinase
Viral Proteins
Issue Date: 2017
Publisher: Genetics Society of America
Citation: Kuhn, L, Majzoub, K, Einhorn, E, Chicher, J, Pompon, J, 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. ScholarBank@NUS Repository. https://doi.org/10.1534/g3.117.042564
Rights: Attribution 4.0 International
Abstract: Receptor 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. © 2017 Kuhn et al.
Source Title: G3: Genes, Genomes, Genetics
URI: https://scholarbank.nus.edu.sg/handle/10635/183516
ISSN: 2160-1836
DOI: 10.1534/g3.117.042564
Rights: Attribution 4.0 International
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