Reconstitution of RNase P activity from inactive RNA and protein.

Abstract

RNase P preparations from Escherichia coli can be separated into RNA and protein by chromatography, in buffers containing 7 M urea, on Sephadex G-200, DEAE-Sephadex or CM-Sephadex columns. Neither RNA nor protein components alone exhibit any RNase activity. RNase P activity can be reconstituted by mixing separated RNA and protein components in buffer containing 7 M urea followed by dialysis of this mixture to remove the urea. Of several purified RNA tried, only M2 RNA, the RNA species found in purified RNase P, is active in the reconstitution experiments.