当前位置：
X-MOL 学术
›
Biochemistry
›
论文详情
Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
Selective Recognition of RNA Substrates by ADAR Deaminase Domains
Biochemistry ( IF 2.9 ) Pub Date : 2018-02-19 00:00:00 , DOI: 10.1021/acs.biochem.7b01100 Yuru Wang 1 , SeHee Park 1 , Peter A. Beal 1
Biochemistry ( IF 2.9 ) Pub Date : 2018-02-19 00:00:00 , DOI: 10.1021/acs.biochem.7b01100 Yuru Wang 1 , SeHee Park 1 , Peter A. Beal 1
Affiliation
|
Adenosine deamination is one of the most prevalent post-transcriptional modifications in mRNA and is catalyzed by ADAR1 and ADAR2 in humans. ADAR1 and ADAR2 have different substrate selectivity, which is believed to mainly originate from the proteins’ deaminase domains (hADAR1d and hADAR2d, respectively). RNA-seq of the Saccharomyces cerevisiae transcriptome subjected to ADAR-catalyzed RNA editing identified substrates with common secondary structure features preferentially edited by hADAR1d over hADAR2d. The relatively small size and efficient reaction of one of these substrates suggested it could be useful for further study of the hADAR1d reaction. Indeed, a short hairpin stem from the S. cerevisiae HER1 mRNA was efficiently deaminated by hADAR1d and used to generate an hADAR1d-specific fluorescent reporter of editing activity. Using substrates preferred by either hADAR1d or hADAR2d in vitro, we found that a chimeric protein bearing an RNA-binding loop from hADAR2d grafted onto hADAR1d showed ADAR2-like selectivity. Finally, a high-throughput mutagenesis analysis (Sat-FACS-Seq) of conserved residues in an RNA-binding loop of hADAR1d revealed essential amino acids for function, advancing our understanding of RNA recognition by this domain.
中文翻译：
通过ADAR脱氨酶域对RNA底物的选择性识别
腺苷脱氨是mRNA中最普遍的转录后修饰之一，在人类中被ADAR1和ADAR2催化。ADAR1和ADAR2具有不同的底物选择性，据信这主要源自蛋白质的脱氨酶结构域（分别为hADAR1d和hADAR2d）。啤酒酵母转录组的RNA-seq接受ADAR催化的RNA编辑后，鉴定出具有常见二级结构特征的底物，其优先被hADAR1d编辑而不是hADAR2d。这些底物之一的相对较小的尺寸和有效的反应表明，它可用于进一步研究hADAR1d反应。确实，啤酒酵母中有一个短发夹HER1 mRNA被hADAR1d有效脱氨基，并用于产生具有编辑活性的hADAR1d特异性荧光报告基因。使用体外受hADAR1d或hADAR2d偏爱的底物，我们发现带有从hADAR2d移植到hADAR1d的RNA结合环的嵌合蛋白显示出ADAR2样的选择性。最后，对hADAR1d RNA结合环中保守残基的高通量诱变分析（Sat-FACS-Seq）显示了功能所需的氨基酸，从而使我们进一步了解了该结构域对RNA的认识。
更新日期：2018-02-19
中文翻译：
通过ADAR脱氨酶域对RNA底物的选择性识别
腺苷脱氨是mRNA中最普遍的转录后修饰之一，在人类中被ADAR1和ADAR2催化。ADAR1和ADAR2具有不同的底物选择性，据信这主要源自蛋白质的脱氨酶结构域（分别为hADAR1d和hADAR2d）。啤酒酵母转录组的RNA-seq接受ADAR催化的RNA编辑后，鉴定出具有常见二级结构特征的底物，其优先被hADAR1d编辑而不是hADAR2d。这些底物之一的相对较小的尺寸和有效的反应表明，它可用于进一步研究hADAR1d反应。确实，啤酒酵母中有一个短发夹HER1 mRNA被hADAR1d有效脱氨基，并用于产生具有编辑活性的hADAR1d特异性荧光报告基因。使用体外受hADAR1d或hADAR2d偏爱的底物，我们发现带有从hADAR2d移植到hADAR1d的RNA结合环的嵌合蛋白显示出ADAR2样的选择性。最后，对hADAR1d RNA结合环中保守残基的高通量诱变分析（Sat-FACS-Seq）显示了功能所需的氨基酸，从而使我们进一步了解了该结构域对RNA的认识。
京公网安备 11010802027423号