Thiolated uridine substrates and templates improve the rate and fidelity of ribozyme-catalyzed RNA copying.
Ribozyme-catalyzed RNA polymerization is inefficient and error prone. Here we demonstrate that two alternative bases, 2-thio-uridine (s(2)U) and 2-thio-ribo-thymidine (s(2)T), improve the rate and fidelity of ribozyme catalyzed nucleotide addition as NTP substrates and as template bases. We also demonstrate the functionality of s(2)U and s(2)T-containing ribozymes.
Howard Hughes Medical Institute, Department of Molecular Biology and Center for Computational and Integrative Biology, Massachusetts General Hospital, 185 Cambridge Street, Boston, Massachusetts 02114, USA and Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford St., Cambridge, Massachusetts 02138, USA.
Noam
N
Prywes
Howard Hughes Medical Institute, Department of Molecular Biology and Center for Computational and Integrative Biology, Massachusetts General Hospital, 185 Cambridge Street, Boston, Massachusetts 02114, USA and Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, University of Oxford, Oxford OX3 9DS, UK.
Yale S
YS
Michaels
Howard Hughes Medical Institute, Department of Molecular Biology and Center for Computational and Integrative Biology, Massachusetts General Hospital, 185 Cambridge Street, Boston, Massachusetts 02114, USA and Department of Genetics, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, Massachusetts 02115, USA. szostak@molbio.mgh.harvard.edu.
Ayan
A
Pal
Howard Hughes Medical Institute, Department of Molecular Biology and Center for Computational and Integrative Biology, Massachusetts General Hospital, 185 Cambridge Street, Boston, Massachusetts 02114, USA and Department of Genetics, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, Massachusetts 02115, USA. szostak@molbio.mgh.harvard.edu.
Seung Soo
SS
Oh
Howard Hughes Medical Institute, Department of Molecular Biology and Center for Computational and Integrative Biology, Massachusetts General Hospital, 185 Cambridge Street, Boston, Massachusetts 02114, USA and Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford St., Cambridge, Massachusetts 02138, USA and Department of Genetics, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, Massachusetts 02115, USA. szostak@molbio.mgh.harvard.edu.
Jack W
JW
Szostak
D055162:Biocatalysis; D007700:Kinetics; D012313:RNA / Q000737:chemistry / Q000235:genetics*; D016337:RNA, Catalytic / Q000737:chemistry / Q000235:genetics*; D048348:Reverse Transcription; D013891:Thiouridine / Q000031:analogs & derivatives* / Q000737:chemistry; D014158:Transcription, Genetic
C105273:2-thiouridine; D016337:RNA, Catalytic; D013891:Thiouridine; C022856:2-thioribothymidine; D012313:RNA