Two helices plus a linker: a small model substrate for eukaryotic RNase P.
Using precursor tRNA molecules to study RNA-protein interactions, we have identified an RNA motif recognized by eukaryotic RNase P (EC 3.1.26.5). Analysis of circularly permuted precursors indicates that interruptions in the sugar-phosphate backbone are not tolerated in the acceptor stem, in the T stem-loop, or between residues A-9 and G-10. Prokaryotic RNase P will function with a minihelix consisting of the acceptor stem connected directly to the T stem-loop. Eukaryotic RNase P cannot use such a minimal substrate unless a linker sequence is added in the gap where the D stem and anticodon stem-loop were deleted.
Carrara
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Istituto di Biologia Cellulare, Consiglio Nazionale delle Ricerche, Rome, Italy.
Calandra
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P
Fruscoloni
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P
Tocchini-Valentini
G P
GP
D000818:Animals; D001483:Base Sequence; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004722:Endoribonucleases / Q000378:metabolism*; D007700:Kinetics; D008961:Models, Structural; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009690:Nucleic Acid Conformation; D012322:RNA Precursors / Q000737:chemistry* / Q000378:metabolism*; D016337:RNA, Catalytic / Q000378:metabolism*; D012360:RNA, Transfer, Phe / Q000737:chemistry* / Q000378:metabolism*; D043262:Ribonuclease P; D013379:Substrate Specificity; D013698:Templates, Genetic; D014158:Transcription, Genetic; D014764:Viral Proteins; D014982:Xenopus laevis
D012322:RNA Precursors; D016337:RNA, Catalytic; D012360:RNA, Transfer, Phe; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; D004722:Endoribonucleases; D043262:Ribonuclease P