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Synthetic substrate analogs for the RNA-editing adenosine deaminase ADAR-2.

机译:RNA编辑腺苷脱氨酶ADAR-2的合成底物类似物。

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摘要

We have synthesized structural analogs of a natural RNA editing substrate and compared editing reactions of these substrates by recombinant ADAR-2, an RNA-editing adenosine deaminase. Deamination rates were shown to be sensitive to structural changes at the 2[prime]-carbon of the edited adenosine. Methylation of the 2[prime]-OH caused a large decrease in deamination rate, whereas 2[prime]-deoxyadenosine and 2[prime]-deoxy-2[prime]-fluoroadenosine were deaminated at a rate similar to adenosine. In addition, a duplex containing as few as 19 bp of the stem structure adjacent to the R/G editing site of the GluR-B pre-mRNA supports deamination of the R/G adenosine by ADAR-2. This identification and initial characterization of synthetic RNA editing substrate analogs further defines structural elements in the RNA that are important for the deamination reaction and sets the stage for additional detailed structural, thermodynamic and kinetic studies of the ADAR-2 reaction.
机译:我们已经合成了天然RNA编辑底物的结构类似物,并通过重组ADAR-2(一种RNA编辑腺苷脱氨酶)比较了这些底物的编辑反应。结果表明,脱氨速率对编辑后的腺苷的2′-碳处的结构变化敏感。 2 [prime] -OH的甲基化导致脱氨速率大大降低,而2 [prime]-脱氧腺苷和2 [prime]-脱氧-2 [prime]-氟代腺苷的脱氨速率与腺苷相似。另外,与GluR-B pre-mRNA的R / G编辑位点相邻的茎结构少至19 bp的双链体支持ADAR-2使R / G腺苷脱氨。合成RNA编辑底物类似物的这种鉴定和初步表征进一步定义了RNA中对脱氨反应重要的结构元素,并为ADAR-2反应的其他详细结构,热力学和动力学研究奠定了基础。

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