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Poster Presentations
Day 4, June 25（Wed.）
Room P (Maesato East, Foyer, Ocean Wing)
- 4P-PM-47
Comprehensive Mapping of Nucleosome-Nuclear Protein Interactions Using Crosslinking Mass Spectrometry
(IQB, UTokyo)
oLumi Negishi, Tomoko Ito, Junko Kato, Tomoya Kujirai, Hitoshi Kurumizaka
This research uses nanoLC-MS/MS proteomics to analyze nucleosome-protein complexes, which regulate vital processes like transcription, replication, and DNA repair. Understanding the molecular recognition of these complexes, often dysregulated in diseases like cancer, requires advanced proteomics techniques. The study employs crosslinking mass spectrometry (crosslinking MS) to identify nuclear proteins interacting with nucleosomes and provides structural insights into nucleosome-nuclear protein complexes. Crosslinking MS helps identify proteins near each other, especially in mobile regions, which are difficult to analyze through other methods like X-ray crystallography. We created MBP-tagged nuclear proteins to map interaction factors in nucleosome-nuclear protein complexes. By mixing these proteins with HeLa cell nuclear extracts and performing pull-down assays, we isolated nucleosome-interacting factors. After crosslinking and proteomic analysis, 654 proteins, including histones, were identified. Interaction mapping revealed 105 protein-protein interactions, with 8 mediated by histones. We are optimizing nuclear protein design, crosslinking strategies, and LC-MS/MS conditions to identify more interaction sites. This presentation highlights our findings from crosslinking MS analyses and our mapping approach for nucleosome-protein complexes.