Template-directed primer extension catalyzed by the Tetrahymena ribozyme.
The Tetrahymena ribozyme has been shown to catalyze an RNA polymerase-like reaction in which an RNA primer is extended by the sequential addition of pN nucleotides derived from GpN dinucleotides, where N = A, C, or U. Here, we show that this reaction is influenced by the presence of a template; bases that can form Watson-Crick base pairs with a template add as much as 25-fold more efficiently than mismatched bases. A mutant enzyme with an altered guanosine binding site can catalyze template-directed primer extension with all four bases when supplied with dinucleotides of the form 2-aminopurine-pN.
Bartel
D P
DP
Department of Molecular Biology, Massachusetts General Hospital, Boston 02114.
Doudna
J A
JA
Usman
N
N
Szostak
J W
JW
D000818:Animals; D001482:Base Composition; D001483:Base Sequence; D001665:Binding Sites; D015226:Dinucleoside Phosphates; D007700:Kinetics; D008969:Molecular Sequence Data; D016337:RNA, Catalytic / Q000378:metabolism*; D013698:Templates, Genetic; D013768:Tetrahymena / Q000235:genetics*
D015226:Dinucleoside Phosphates; D016337:RNA, Catalytic