Escherichia coli DbpA is an RNA helicase that requires hairpin 92 of 23S rRNA

Escherichia coli DbpA is a member of the DEAD/H family of proteins which ha s been shown to have robust ATPase activity only in the presence of a speci fic region of 23S rRNA. A series of bimolecular RNA substrates were designe d based on this activating region of rRNA and used to demonstrate that DbpA is also a non-processive, sequence-specific RNA helicase. The high affinit y of DbpA for the RNA substrates allowed both single and multiple turnover helicase assays to be performed. Helicase activity of DbpA is dependent on the presence of ATP or dATP, the sequence of the loop of hairpin 92 of 23S rRNA and the position of the substrate helix with respect to hairpin 92. Th is work indicates that certain RNA helicases require particular RNA structu res in order for optimal unwinding activity to be observed.