选择性聚合酶的识别使得在RNA中检测到N(6) - - 甲基氨酸
Emily M Harcourt1, Thomas Ehrenschwender, Pedro J Batista
1Department of Chemistry, Stanford University , Stanford, California 94305, United States.
Journal of the American Chemical Society
|December 17, 2013
概括
研究人员发现了一种新型酶,可以精确地定位N(6) - 甲基氨酸 (m(6) A) RNA修饰. 这一突破确定了m(6) A在哺乳动物28S rRNA中的核酸4190,推动了RNA研究和人类健康研究.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 表观遗传学 在表观遗传学中,表观遗传学是指表观遗传学.
背景情况:
- 甲基氨酸 (M6A) 是真核生物中最常见的mRNA修饰.
- m(6) A在各种生物过程和人类健康中起着至关重要的作用.
- 精确地定位RNA序列中的m(6) A修饰仍然是一个重大的技术挑战.
研究的目的:
- 开发一种方法来精确地定位和量化RNA中的m(6) A修饰.
- 确定一种能够区分改性和未改性腺残留物的特定酶.
- 在哺乳动物28S核糖体RNA (rRNA) 中确定m(6) A的确切位置.
主要方法:
- 从*Thermus thermophilus*获得的具有逆转录酶活性的聚合酶的查和鉴定.
- 酶选择性的特征,用于在m(6)A.A.的时间内将提米丁与未经修改的腺素相对应.
- 选择性聚合酶与非选择性聚合酶一起用于合成和细胞RNA中的m(6) A映射的应用.
主要成果:
- 已确定的*Thermus thermophilus*聚合酶对未经修改的腺具有多达18倍的选择性.
- 该酶可以通过对暂停带的分析在合成RNA中定位和量化m(6) A.
- 该研究成功地确定了哺乳动物28S rRNA中m(6) A的精确位置,位于核酸4190.0.
结论:
- 已经建立了一种新的酶方法,用于精确地绘制m(6) A修饰的图表.
- 这种方法克服了在RNA分子内识别特定的m(6) A位置方面的先前限制.
- 在哺乳动物28S rRNA中的核酸4190中确定m(6) A为RNA调节和功能提供了关键的见解.
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