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Serial interactome capture of the human cell nucleus
Thomas Conrad1, Anne-Susann Albrecht1,2, Veronica Rodrigues de Melo Costa1,3
1Max Planck Institute for Molecular Genetics, Otto Warburg Laboratories, 14195 Berlin, Germany.
Nature Communications
|April 5, 2016
Summary
We developed serial RNA interactome capture (serIC) to identify nuclear RNA-binding proteins (RBPs). This method revealed a novel network of RBPs involved in DNA repair and p53 signaling.
Area of Science:
- Molecular Biology
- Genomics
- Cell Biology
Background:
- RNA-binding proteins (RBPs) play crucial roles in cellular functions.
- Understanding the full spectrum of RBPs is essential for comprehending gene regulation.
Purpose of the Study:
- To introduce a novel method, serial RNA interactome capture (serIC), for high-specificity global detection of RBPs.
- To identify the human nuclear RNA interactome in proliferating K562 cells.
Main Methods:
- Serial RNA interactome capture (serIC) involves multiple purification steps of UV-crosslinked poly(A)-RNA-protein complexes.
- Application of serIC to nuclei of proliferating K562 cells.
Main Results:
- Identified 382 nuclear RBPs, forming the first human nuclear RNA interactome.
- Observed distinct domain composition of identified RBPs compared to previous methods.
- Discovered novel DNA-RNA-binding proteins (DRBPs) and a network of RBPs implicated in p53 signaling and double-strand break repair.
Conclusions:
- serIC is an effective tool for global RBP detection with high specificity.
- The identified nuclear RBP network provides new insights into DNA repair and p53 signaling pathways.
- serIC facilitates further specific detection of purified RBPs through additional selection or labeling.
