Measuring Helicase Inhibition of the DEAD-Box Protein Dbp2 by Yra1
Despite the highly conserved helicase core, individual DEAD-box proteins are specialized in diverse RNA metabolic processes. One mechanism that determines DEAD-box protein specificity is enzymatic regulation by other protein cofactors. In this chapter, we describe a protocol for purifying the Saccharomyces cerevisiae DEAD-box RNA helicase Dbp2 and RNA-binding protein Yra1 and subsequent analysis of helicase regulation. The experiments described here can be adapted to other RNA helicases and their purified cofactor(s).
Key wordsDEAD-box RNA Helicase Unwinding Annealing Duplex Yeast
We thank members of the Tran lab for constructive criticism and detailed analysis of experimental methods. This work was supported by NIH grant R01GM097332 to E.J.T. and by P30 CA023168.
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