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The Effect of Cytidine on the Structure and Function of an RNA Ligase RibozymeA cytidine-free ribozyme with RNA ligase activity was obtained by in vitro evolution, starting from a pool of random- sequence RNAs that contained only guanosine, adenosine, and uridine. This ribozyme contains 74 nt and catalyzes formation of a 3',5' -phosphodiester linkage with a catalytic rate of 0.016/min. The RNA adopts a simple secondary structure based on a three-way junction motif, with ligation occurring at the end of a stem region located several nucleotides away from the junction. Cytidine was introduced to the cytidine-free ribozyme in a combinatorial fashion and additional rounds of in vitro evolution were carried out to allow the molecule to adapt to this added component. The resulting cytidine-containing ribozyme formed a 3',5' linkage with a catalytic rate of 0.32/min. The improved rate of the cytidine-containing ribozyme was the result of 12 mutations, including seven added cytidines, that remodeled the internal bulge loops located adjacent to the three-way junction and stabilized the peripheral stem regions.
Document ID
20030066911
Acquisition Source
Headquarters
Document Type
Reprint (Version printed in journal)
Authors
Rogers, Jeff
(Scripps Research Inst. La Jolla, CA, United States)
Joyce, Gerald F.
(Scripps Research Inst. La Jolla, CA, United States)
Date Acquired
August 21, 2013
Publication Date
January 1, 2001
Publication Information
Publication: RNA
Publisher: Cambridge Univ. Press
Volume: 7
Subject Category
Exobiology
Funding Number(s)
CONTRACT_GRANT: NAG5-3647
CONTRACT_GRANT: NAG5-4546
Distribution Limits
Public
Copyright
Other

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