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

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.

SUBMITTER: Spiniello M

PROVIDER: S-EPMC6800478 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature

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

Spiniello Michele M Steinbrink Maisie I MI Cesnik Anthony J AJ Miller Rachel M RM Scalf Mark M Shortreed Michael R MR Smith Lloyd M LM

RNA (New York, N.Y.) 20190711 10

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-M ...[more]

PMID: 31296583

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