一个与DYW1相关的核糖酶活性在体外被RIP/MORF蛋白抑制
Robert D Boyd1, Michael L Hayes2
1Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA.
Scientific reports
|July 3, 2023
概括
植物RNA编辑因子DYW1在体外表现出核糖酶活性,其被其他编辑因子抑制. 这表明,原生编辑复杂物在体内调节和限制DYW1的核酶活性.
科学领域:
- 植物分子生物学 植物分子生物学
- RNA生物化学与RNA生物化学
- 酶学 是一种酶学.
背景情况:
- 在植物中,有机细胞C-to-URNA编辑依赖于核编码的蛋白质复合体.
- DYW-deaminases是C-to-URNA编辑中的关键酶,但有些在体外表现出不典型的核糖酶活性.
- 这种核糖核酶活动对于去除毒素并非必不可少,并且可能对mRNA编辑有害.
研究的目的:
- 为了研究植物DYW1编辑因子的核糖酶活性.
- 确定离子在这种活动中的作用.
- 评估其他RNA编辑因子对DYW1核酶活性的影响.
主要方法:
- 使用固定金属亲属性染色学净化复合的阿拉比多普西斯塔利亚纳DYW1 (rAtDYW1).
- 在各种条件下,测试 rAtDYW1 在光标记的 RNA 寡核酸上对核糖酶活性.
- 检查化剂和其他纯化重组编辑因子 (RIP/MORF蛋白,AtCRR4,AtORRM1,AtOZ1) 对rAtDYW1活性的影响.
主要成果:
- 纯化rAtDYW1在体外证明了RNA裂变 (水解).
- 分裂活动对化剂敏感,这表明依赖的机制.
- 与RIP/MORF蛋白共同化降低了AtDYW1的核糖酶活性.
- 虽然AtCRR4,AtORRM1和AtOZ1没有强烈抑制缺乏特定cis元素的RNA的活性,但AtCRR4抑制了具有相关cis元素的RNA的活性.
结论:
- 观察到的RNA裂变与重组AtDYW1.1内在相关.
- 离子在rAtDYW1.1.的核酶活性中起作用.
- 其他RNA编辑因子的存在,特别是RIP/MORF蛋白和AtCRR4 (与其cis元素),可以抑制DYW1.1的核糖酶活性.
- 这些发现表明,原生编辑复杂物调节DYW1的核酶活性,在体内潜在地防止非目标RNA降解.
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