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Abstract:
Novel RNA-guided cellular functions are paralleled by an increasing number of RNA-binding
proteins (RBPs). Here we present ‘serial RNA interactome capture’ (serIC), a multiple
purification procedure of ultraviolet-crosslinked poly(A)–RNA–protein complexes that
enables global RBP detection with high specificity. We apply serIC to the nuclei of
proliferating K562 cells to obtain the first human nuclear RNA interactome. The domain
composition of the 382 identified nuclear RBPs markedly differs from previous IC
experiments, including few factors without known RNA-binding domains that are in good
agreement with computationally predicted RNA binding. serIC extends the number of
DNA–RNA-binding proteins (DRBPs), and reveals a network of RBPs involved in p53 signalling
and double-strand break repair. serIC is an effective tool to couple global RBP capture with
additional selection or labelling steps for specific detection of highly purified RBPs.
