Synthesis and evaluation of an RNA editing substrate bearing 2′-deoxy-2′-mercaptoadenosine

Prasanna Jayalath, Subhash Pokharel, Eduardo Véliz, Peter A. Beal

Research output: Contribution to journalArticle

5 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)78-88
Number of pages11
JournalNucleosides, Nucleotides and Nucleic Acids
Volume28
Issue number2
DOIs
StatePublished - Feb 2009

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Keywords

  • ADARs
  • Nucleotide binding
  • RNA substrates

ASJC Scopus subject areas

  • Genetics
  • Biochemistry
  • Molecular Medicine

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