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Published on: July 22, 2014
Synthesis and evaluation of an RNA editing substrate bearing 2'-deoxy-2'-mercaptoadenosine
Prasanna Jayalath1, Subhash Pokharel, Eduardo Véliz
1Department of Chemistry, University of California Davis, California 95616, USA.
Abstract:
The RNA-editing adenosine deaminases (ADARs) catalyze deamination of adenosine to inosine in double stranded structure found in various RNA substrates, including mRNAs. Here we describe the synthesis of a phosphoramidite of 2'-deoxy-2'-mercaptoadenosine and its incorporation into an ADAR substrate. Surprisingly, no deamination product was observed with this substrate indicating replacing the 2'-OH with a 2'-SH at the editing site is highly inhibitory. Modeling of nucleotide binding into the active site suggests the side chain of T375 of human ADAR2 to be in proximity of the 2'-substituent. Mutation of this residue to cysteine caused a greater that 100-fold reduction in deamination rate with the 2'-OH substrate.
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