期刊
NUCLEIC ACIDS RESEARCH
卷 37, 期 9, 页码 3074-3082出版社
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkp177
关键词
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资金
- W. M. Keck Foundation
- National Science Foundation [MCB 0543741]
New classes of RNA enzymes or ribozymes have been obtained by in vitro evolution and selection of RNA molecules. Incorporation of modified nucleotides into the RNA sequence has been proposed to enhance function. DA22 is a modified RNA containing 5-(4-pyridylmethyl) carboxamide uridines, which has been selected for its ability to promote a DielsAlder cycloaddition reaction. Here, we show that DATR96, the most active member of the DA22 RNA sequence family, which was selected with pyridyl-modified nucleotides, accelerates a cycloaddition reaction between anthracene and maleimide derivatives with high turnover. These widely used reactants were not used in the original selection for DA22 and yet here they provide the first demonstration of DATR96 as a true multiple-turnover catalyst. In addition, the absence of a structural or essential kinetic role for Cu-2, as initially postulated, and nonsequence-specific hydrophobic interactions with the anthracene substrate have led to a reevaluation of the pyridine modifications role. These findings broaden the catalytic repertoire of the DA22 family of pyridyl-modified RNAs and suggest a key role for the hydrophobic effect in the catalytic mechanism.
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