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クロマチンプロテオミクスChEPによるインスリン依存的な転写システムの解明
Chromatin is a large complex of DNA and proteins which form chromatin within the nucleus of eukaryotic cells. The primary functions of chromatin are packaging DNA into a more compact shape, controlling gene expression and DNA replication. These biological processes are selectively mediated by the different set of proteins under particular stimuli. Therefore, comprehensive identification of chromatin-associated proteins provides useful information to understand how cells respond to extracellular stimuli via chromatin reorganization. Recently, Kustatscher et al. developed Chromatin Enrichment For Proteomics (ChEP), which is a simple biochemical procedure that enriches interphase chromatin in all its complexity. This approach enables efficient identification and quantification of chromatin proteins.
Here, we investigated how chromatin proteins are orchestrally regulated at a proteome-wide scale by using Fao hepatoma cells stimulated with insulin. We checked dynamics of chromatin proteins after insulin stimulation with a combination of ChEP and SWATH-MS analysis, a data independent acquisition (DIA) method. We identified insulin-regulated chromatin proteins, which showed a significant increase or decrease after insulin stimulation. Of these proteins, splicing factors, such as SRSF1, and SRRM1, decreased transiently after insulin stimulation. These results demonstrate that insulin signaling activates or inhibits not only transcription factors, but also splicing factors to regulate transcription.