Predicting sites of ADAR editing in double-stranded RNA.

Predicting sites of ADAR editing in double-stranded RNA.
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DOI:
10.1038/ncomms1324
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发表时间:
2011
影响因子:
16.6
通讯作者:
Bass, Brenda L.
Bass, Brenda L.
中科院分区:
综合性期刊1区
文献类型:
--
作者:
Eggington, Julie M.;Greene, Tom;Bass, Brenda L.

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ADAR(作用于 RNA 的腺苷脱氨酶)编辑酶以编码和非编码双链 RNA (dsRNA) 为目标,对神经元功能至关重要。早期研究表明 ADAR 优先靶向具有某些 5' 和 3' 邻居的腺苷。在这里,我们使用当前的桑格测序方案来进行更准确和定量的分析。在与人 ADAR1 或 ADAR2 或其单独的催化结构域反应后,我们对~800 bp dsRNA 中的编辑位点进行了定量。这些大型数据集表明,邻居偏好主要由催化结构域决定,但 ADAR2 的 dsRNA 结合基序有助于 3' 邻居偏好。对于所有蛋白质,5'最近邻居影响最大,但相邻碱基也会影响编辑位点的选择。我们开发了算法来预测任何序列的 dsRNA 中的编辑位点,并提供基于网络的应用程序。与包含错配、凸起和环的生物底物相比,该算法对完全碱基配对的 dsRNA 的预测能力阐明了结构对编辑特异性的贡献。 ADAR 酶编辑双链 RNA,将腺苷转化为肌苷,对于神经元功能至关重要。埃金顿等人。使用 Sanger 测序方案量化 RNA 中的编辑位点,并使用所得数据开发算法来预测 RNA 编辑位点。
ADAR (adenosine deaminase that acts on RNA) editing enzymes target coding and noncoding double-stranded RNA (dsRNA) and are essential for neuronal function. Early studies showed that ADARs preferentially target adenosines with certain 5′ and 3′ neighbours. Here we use current Sanger sequencing protocols to perform a more accurate and quantitative analysis. We quantified editing sites in an ∼800-bp dsRNA after reaction with human ADAR1 or ADAR2, or their catalytic domains alone. These large data sets revealed that neighbour preferences are mostly dictated by the catalytic domain, but ADAR2's dsRNA-binding motifs contribute to 3′ neighbour preferences. For all proteins, the 5′ nearest neighbour was most influential, but adjacent bases also affected editing site choice. We developed algorithms to predict editing sites in dsRNA of any sequence, and provide a web-based application. The predictive power of the algorithm on fully base-paired dsRNA, compared with biological substrates containing mismatches, bulges and loops, elucidates structural contributions to editing specificity. ADAR enzymes edit double-stranded RNA, converting adenosines to inosines, and are essential for neuronal function. Eggington et al. quantify edit sites in RNA using a Sanger sequencing protocol and use the resulting data to develop algorithms to predict RNA edit sites.
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