Selection of guide sequences that direct efficient cleavage of mRNA by human ribonuclease P.
Any RNA, when in a complex with another oligoribonucleotide known as an external guide sequence (EGS), can become a substrate for ribonuclease P. Simulation of evolution in vitro was used to select EGSs that bind tightly to a target substrate messenger RNA and that increase the efficiency of cleavage of the target by human ribonuclease P to a level equal to that achieved with natural substrates. The most efficient EGSs form transfer RNA precursor-like structures with the target RNA, in which the analog of the anticodon stem has been disrupted, an indication that selection for the optimal substrate for ribonuclease P yields an RNA structure different from that of present-day transfer RNA precursors.
Yuan
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Y
Department of Biology, Yale University, New Haven, CT 06520.
Altman
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S
D000926:Anticodon / Q000737:chemistry / Q000378:metabolism; D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D004722:Endoribonucleases / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009843:Oligoribonucleotides / Q000737:chemistry / Q000378:metabolism*; D012322:RNA Precursors / Q000737:chemistry / Q000378:metabolism; D016337:RNA, Catalytic / Q000378:metabolism*; D012333:RNA, Messenger / Q000737:chemistry / Q000378:metabolism*; D012343:RNA, Transfer / Q000737:chemistry / Q000378:metabolism; D043262:Ribonuclease P; D013816:Thermodynamics; D058727:RNA, Small Untranslated
D000926:Anticodon; D009843:Oligoribonucleotides; D012322:RNA Precursors; D016337:RNA, Catalytic; D012333:RNA, Messenger; D012343:RNA, Transfer; D015500:Chloramphenicol O-Acetyltransferase; D004722:Endoribonucleases; C468368:RPP14 protein, human; D043262:Ribonuclease P; D008274:Magnesium; D058727:RNA, Small Untranslated