Structure and specific RNA binding of ADAR2 double-stranded RNA binding motifs
Structure and specific RNA binding of ADAR2 double-stranded RNA binding motifs
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DOI：
10.1016/j.str.2005.11.013
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发表时间：
2006-02-01
期刊：
影响因子：
5.7
通讯作者：
Allain, FHT
中科院分区：
文献类型：
--
作者：
Stefl, R;Xu, M;Allain, FHT
作用于RNA(ADAR)位点的腺苷脱氨酶选择性地将RNA转录本中的腺苷修饰为肌苷，从而重新编码基因组信息。ADAR如何选择特定的腺苷部分进行脱氨还知之甚少。在这里，我们报道了大鼠ADAR2的两个双链RNA结合基序(DsRBMs)的核磁共振结构，并对这两个dsRBMs与编码GluR-B R/G位点的71个核苷酸RNA的相互作用进行了核磁共振化学位移微扰研究。我们已经确定了参与复合体形成的蛋白质和RNA表面，使我们能够提供一个基于核磁共振的复合体模型。我们发现dsRBM1识别保守的五元环，而dsRBM2识别编辑位点附近的两个凸起的碱基，表明ADAR2 dsRBM对RNA结构的识别。蛋白质和RNA的体外突变研究进一步支持了我们的结构发现。
Adenosine deaminases that act on RNA (ADARs) siteselectively modify adenosines to inosines within RNA transcripts, thereby recoding genomic information. How ADARs select specific adenosine moieties for deamination is poorly understood. Here, we report NMR structures of the two double-stranded RNA binding motifs (dsRBMs) of rat ADAR2 and an NMR chemical shift perturbation study of the interaction of the two dsRBMs with a 71 nucleotide RNA encoding the R/G site of the GluR-B. We have identified the protein and the RNA surfaces involved in complex formation, allowing us to present an NMR-based model of the complex. We have found that dsRBM1 recognizes a conserved pentaloop, whereas dsRBM2 recognizes two bulged bases adjacent to the editing site, demonstrating RNA structure-dependent recognition by the ADAR2 dsRBMs. In vitro mutagenesis studies with both the protein and the RNA further support our structural findings.