Comprehensive in vivo identification of the c-Myc mRNA protein interactome using HyPR-MS

使用 HyPR-MS 全面体内鉴定 c-Myc mRNA 蛋白质相互作用组

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作者:Michele Spiniello #, Maisie I Steinbrink #, Anthony J Cesnik, Rachel M Miller, Mark Scalf, Michael R Shortreed, Lloyd M Smith

Abstract

Proteins bind mRNA through their entire life cycle from transcription to degradation. We analyzed c-Myc mRNA protein interactors in vivo using the HyPR-MS method to capture the crosslinked mRNA by hybridization and then analyzed the bound proteins using mass spectrometry proteomics. Using HyPR-MS, 229 c-Myc mRNA-binding proteins were identified, confirming previously proposed interactors, suggesting new interactors, and providing information related to the roles and pathways known to involve c-Myc. We performed structural and functional analysis of these proteins and validated our findings with a combination of RIP-qPCR experiments, in vitro results released in past studies, publicly available RIP- and eCLIP-seq data, and results from software tools for predicting RNA-protein interactions.

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