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非正规的二次结构通过降低三级折叠的热性成本来稳定线粒体tRNA (((Ser (((UCN))
Anthony M Mustoe, Xin Liu, Paul J Lin
1∥Departments of Biochemistry and Chemistry, Duke University School of Medicine, Durham, North Carolina 27710, United States.
Journal of the American Chemical Society
|February 24, 2015
概括
线粒体tRNA (mt-tRNA) 的拓约束抵消了失去的三级相互作用,稳定了它的结构. 破坏这些约束的致病突变使mt-tRNA不稳定,将其与聋和等疾病联系起来.
科学领域:
- 分子生物学分子生物学
- 生物物理学的生物物理.
- 遗传学 遗传学 是一个
背景情况:
- 哺乳动物线粒体tRNAs (mt-tRNAs) 和pyrrolysine tRNAs (tRNA(Pyl)) 采用复杂的3D结构,尽管异常的二次结构破坏了典型的tRNA三级相互作用.
- 这些非正规的tRNA保持稳定的机制以及突变对它们功能的影响仍然在很大程度上是未知的.
研究的目的:
- 为了研究非正规的tRNA如何补偿中断的三级相互作用.
- 阐明病原性7472insC突变在人类mt-tRNA中的分子机制及其与神经系统疾病的关联.
主要方法:
- 利用TOPRNA粗粒度模拟来模拟RNA折叠和拓约束.
- 进行紫外线化实验,以实验验证关于tRNA稳定性和折叠自由能量的模拟预测.
主要成果:
- 野生类型mt-tRNA的拓约束增加降低了折叠成本,补偿了失去的三级相互作用.
- 病原性7472insC突变被证明会破坏拓约束,使mt-tRNA ((Ser) 不稳定大约0.6kcal/mol.
- 实验数据证实,突变降低了融温度,增加了折叠的自由能量,与模拟结果一致.
结论:
- 拓约束对于非正规tRNA的稳定性至关重要,为RNA折叠提供了定量框架.
- 7472insC突变代表了第一个由拓约束的破坏引起的确定的致病突变,为疾病机制提供了洞察力.
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