Probing adenosine-to-inosine editing reactions using RNA-containing nucleoside analogs.

Abstract:

:Advances in chemical synthesis and characterization of nucleic acids allows for atom-specific modification of complex RNAs, such as present in RNA editing substrates. By preparing substrates for ADARs by chemical synthesis, it is possible to subtly alter the structure of the edited nucleotide. Evaluating the effect these changes have on the rate of enzyme-catalyzed deamination reveals features of the editing reaction and guides the design of inhibitors. We describe the synthesis of select nucleoside analog phosphoramidites and their incorporation into RNAs that mimic known editing sites by solid phase synthesis, and analyze the interaction of these synthetic RNAs with ADARs using deamination kinetics and quantitative gel mobility shift assays.

journal_name

Methods Enzymol

journal_title

Methods in enzymology

authors

Maydanovych O,Easterwood LM,Cui T,Véliz EA,Pokharel S,Beal PA

doi

10.1016/S0076-6879(07)24017-0

subject

Has Abstract

pub_date

2007-01-01 00:00:00

pages

369-86

eissn

0076-6879

issn

1557-7988

pii

S0076-6879(07)24017-0

journal_volume

424

pub_type

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