Purified protein was mixed with [32]P labeled pre-miRNA. Binding was monitored by gel shift.
Interaction in vitro; bait produced as a recombinant fusion protein in baculovirus-infected insect cell system; prey produced and labeled by in vitro transcription.
Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey produced and labeled by in vitro transcription.
Interaction in vitro; RNA produced and labeled by in vitro transcription; protein produced as a recombinant fusion protein in an S2 cell system.
Binding affinity is greatest with at least 14nt of single-stranded RNA at loop end of substrate, at most 22nt of double-stranded RNA and a 2nt overhang at stem end. Binding affinity unaffected by addition of ATP.
