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In Vitro RNA-Protein Binding Assays to Identify RNA Helicases for U snRNA Nuclear Export
1Graduate School of Frontier Biosciences, The University of Osaka, Suita, Japan. taniguchi.ichiro.fbs@osaka-u.ac.jp.
Abstract:
RNA helicases play a critical role in RNA biogenesis by modulating RNA-protein interactions in an ATP-dependent manner. Although many RNA helicases function by unwinding RNA secondary structures or displacing proteins from RNA, a subset of RNA helicases facilitates the loading of their interacting proteins onto RNA. Here, I present an in vitro method for analyzing the formation of the uridine-rich small nuclear RNA (U snRNA) export complex, for which the involvement of RNA helicases was previously unknown. This method can be used to determine whether complex formation is ATP-dependent and identify RNA helicases that facilitate the loading of the U snRNA export factor onto RNA. This approach can identify RNA helicases involved in protein loading onto various RNA-protein complexes.