Substrate recognition by RNase P and by the catalytic M1 RNA: identification of possible contact points in pre-tRNAs.
Modified bases were introduced into pre-tRNAs during in vitro RNA synthesis or by chemical modification. These RNAs were used as substrates for the catalytic M1 RNA and the RNase P holoenzyme from Schizosaccharomyces pombe. The synthetic approach permitted the insertion of 100% m7GTP into pre-tRNAs and this resulted in complete inhibition of the specific 5' processing reactions. Partially modified RNAs were obtained by chemical modifications of purines and uridines in the pre-tRNAs. This allowed detailed analyses of specific bases excluded in the products. With pre-tRNA(Ser) and initiator pre-tRNA(Met), strong effects were observed in the T arm and weaker effects in the anticodon stem. Only minor base exclusions were detected in the acceptor stem of pre-tRNA(Ser) and in the D arm of pre-tRNA(Met).
Kahle
D
D
Institute for General Microbiology, Christian-Albrechts-Universität, Kiel, FRG.
Wehmeyer
U
U
Krupp
G
G
D001483:Base Sequence; D001665:Binding Sites; D004722:Endoribonucleases / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012322:RNA Precursors / Q000032:analysis*; D012323:RNA Processing, Post-Transcriptional; D012343:RNA, Transfer / Q000032:analysis*; D043262:Ribonuclease P; D004718:Saccharomycetales / Q000235:genetics*; D012568:Schizosaccharomyces / Q000201:enzymology / Q000235:genetics*; D013379:Substrate Specificity
D012322:RNA Precursors; D012343:RNA, Transfer; D004722:Endoribonucleases; D043262:Ribonuclease P