(A) Schematic depiction of human RNase P RNA and deletion fragments.
(B) Import of full length RNase P RNA into yeast mitochondria expressing PNPASE (PNP) or control (Vec) vectors, as in Figure 4A.
(C) Import of the indicated RNase P RNA fragments.
(D) Import of RNase P RNA fragments RPf3 and RPf4.
(E) Import of human GAPDH mRNA or GAPDH mRNA with control (CR), MRP RNA, or RNase P RNA 20 nt sequences fused to the 5’ end, as shown in panel F.
(F) The secondary structures and sequences of mitochondrial RNA targeting signals in RNase P (RP) and MRP (MRP) RNAs. A random sequence (CR) was used as a control.
(G) Isolated mitochondria from HEK293 cells stably expressing IMS-localized PNPASE-HisPC or TIM23-HisPC (control) dual-tagged proteins were incubated with [32P]-CTP labeled CR-tRNAtrp or RP-tRNAtrp, followed by UV-cross linking, tag-IP, separation by SDS-PAGE, and autoradiography. See also Figure S6.