Abstract
We have discovered an artificial DNA enzyme that mimics the first step of RNA splicing. In vitro selection was used to identify DNA enzymes that ligate RNA. One of the new DNA enzymes carries out splicing-related catalysis by specifically recognizing an unpaired internal adenosine and facilitating attack of its 2′-hydroxyl onto a 5′-triphosphate. This reaction forms 2′,5′-branched RNA and is analogous to the first step of in vivo RNA splicing, in which a ribozyme cleaves itself with formation of a branched intermediate. Unlike a natural ribozyme, the new DNA enzyme has no 2′-hydroxyl groups to aid in the catalytic mechanism. Our finding has two important implications. First, branch-site adenosine reactivity seems to be mechanistically favored by nucleic acid enzymes. Second, hydroxyl groups are not obligatory components of nucleic acid enzymes that carry out biologically related catalysis.
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Acknowledgements
This research was supported by the Burroughs Wellcome Fund (New Investigator Award in the Basic Pharmacological Sciences), the March of Dimes Birth Defects Foundation (Basil O'Connor Starter Scholar Research Award), the US National Institutes of Health, the Petroleum Research Fund administered by the American Chemical Society, and the University of Illinois at Urbana-Champaign Department of Chemistry (all to S.K.S.). S.K.S. is the recipient of a fellowship from The David and Lucile Packard Foundation. We thank members of the Silverman lab for discussions, H. Imker for technical assistance with the background reaction rate experiments, Y. Wang for testing lariat RNA formation with 7S11 and P. Hergenrother and J. Cottell for comments on the manuscript.
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Coppins, R., Silverman, S. A DNA enzyme that mimics the first step of RNA splicing. Nat Struct Mol Biol 11, 270–274 (2004). https://doi.org/10.1038/nsmb727
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DOI: https://doi.org/10.1038/nsmb727
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