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通过冷-EM捕获可巴拉明 рибо开关的异质适配体
Jienyu Ding1, Justin C Deme2, Jason R Stagno1
1Protein-Nucleic Acid Interaction Section, Center for Structural Biology, Center for Cancer Research, National Cancer Institute, Frederick, MD 21702, USA.
Nucleic acids research
|August 3, 2023
概括
连接物结合稳定了腺赛尔科巴胺 рибо开关,降低了其结构灵活性. 这项研究确定了冷-EM结构,揭示了对RNA功能至关重要的联结体诱导的变化.
科学领域:
- 结构生物学 结构生物学
- 分子生物学分子生物学
- 生物化学 生物化学
背景情况:
- 对于高分辨率的结构确定,特别是对于大型灵活的RNA来说,RNA结构异质性带来了挑战.
- 已知adenosylcobalamin riboswitch表现出结构灵活性,受Mg2+度和连接体结合的影响.
研究的目的:
- 为了阐明基诱导的形状变化的结构基础在adenosylcobalamin riboswitch.
- 为了确定高分辨率的冷电子显微镜结构的riboswitch的apo和全息状态.
主要方法:
- 使用冷电子显微镜 (cryo-EM) 确定了带丝切换器的结构.
- 进行了骨干比较和对Apo和Holo状态的分析.
主要成果:
- 确定了一个apo和四个全息冷EM结构,全息结构分辨率为3.0-3.5 Å.
- 联体结合显著降低了形状灵活性.
- 在P6延伸和结合口袋中观察到显著的结构差异,结合点6/3采用"S"到"U"形状的形状,以适应连接体.
- 结合口袋的形成取决于Mg2+度,在1mM时部分形成,在10mM时完全形成Mg2+.
结论:
- 连接物结合会诱导腺赛尔科巴胺 рибо开关中的稳定形状变化.
- P6延伸在连接体结合和选择性中起作用.
- 了解这些结构动态,可以深入了解 рибо开关调节机制.
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