Proteomic Analysis of the EWS-Fli-1 Interactome Reveals the Role of the Lysosome in EWS-Fli-1 Turnover.
Ewing sarcoma is a cancer of bone and soft tissue in children that is characterized by a chromosomal translocation involving EWS and an Ets family transcription factor, most commonly Fli-1. EWS-Fli-1 fusion accounts for 85% of cases. The growth and survival of Ewing sarcoma cells are critically dependent on EWS-Fli-1. ... A large body of evidence has established that EWS-Fli-1 functions as a DNA-binding transcription factor that regulates the expression of a number of genes important for cell proliferation and transformation. However, little is known about the biochemical properties of the EWS-Fli-1 protein. We undertook a series of proteomic analyses to dissect the EWS-Fli-1 interactome. Employing a proximity-dependent biotinylation technique, BioID, we identified cation-independent mannose 6-phosphate receptor (CIMPR) as a protein located in the vicinity of EWS-Fli-1 within a cell. CIMPR is a cargo that mediates the delivery of lysosomal hydrolases from the trans-Golgi network to the endosome, which are subsequently transferred to the lysosomes. Further molecular cell biological analyses uncovered a role for lysosomes in the turnover of the EWS-Fli-1 protein. We demonstrate that an mTORC1 active-site inhibitor, torin 1, which stimulates the TFEB-lysosome pathway, can induce the degradation of EWS-Fli-1, suggesting a potential therapeutic approach to target EWS-Fli-1 for degradation.
Mesh Terms:
Biotinylation, Catalytic Domain, Cell Line, Tumor, Cell Proliferation, Cell Transformation, Neoplastic, Gene Expression Regulation, Neoplastic, HEK293 Cells, HeLa Cells, Humans, Lysosomes, Multiprotein Complexes, Oncogene Proteins, Fusion, Proteome, Proteomics, Proto-Oncogene Protein c-fli-1, RNA-Binding Protein EWS, Sarcoma, Ewing, TOR Serine-Threonine Kinases, Tandem Mass Spectrometry, Transcription Factors, trans-Golgi Network
Biotinylation, Catalytic Domain, Cell Line, Tumor, Cell Proliferation, Cell Transformation, Neoplastic, Gene Expression Regulation, Neoplastic, HEK293 Cells, HeLa Cells, Humans, Lysosomes, Multiprotein Complexes, Oncogene Proteins, Fusion, Proteome, Proteomics, Proto-Oncogene Protein c-fli-1, RNA-Binding Protein EWS, Sarcoma, Ewing, TOR Serine-Threonine Kinases, Tandem Mass Spectrometry, Transcription Factors, trans-Golgi Network
J. Proteome Res.
Date: Aug. 01, 2014
PubMed ID: 24999758
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