四合体组I内的反向自我拼接:对拼接的方向性和内转位的含义
1Howard Hughes Medical Institute, Department of Chemistry and Biochemistry, University of Colorado, Boulder 80309-0215.
Cell
|April 21, 1989
概括
泰特拉海门 rRNA 组 I 内子.
科学领域:
- 分子生物学分子生物学
- 在RNA分离过程中.
- 生物化学 生物化学
背景情况:
- I 组的内子是自我拼接的催化RNA.
- 甲基 rRNA I 组内是研究 RNA 拼接机制的模型系统.
研究的目的:
- 为了研究 Tetrahymena rRNA 组 I 内子拼接 in vitro 的可逆性.
- 确定影响RNA拼接方向性的因素.
- 探索内部集成对转换的影响.
主要方法:
- 在体外拼接试验中,使用短的寡核酸作为结合的外子基质.
- 在不同的条件下 (RNA度,,温度,瓜诺辛存在) 用线性内子化结合的外因子RNA.
- 对外子序列要求和二次结构在拼接中的作用的分析.
主要成果:
- 在体外 (in vitro) 中,四胺rRNA组I内拼接是完全可逆的.
- 由于高RNA度,,温度和瓜诺辛的缺乏,逆转更容易发生.
- 与内部导向序列配对的5'外显子序列是必不可少的;3'外显子序列不是.
- 与非结构化基板相比,内部集成到结构化基板中显著减少.
- 这种Tetrahymena内可以整合到β-环球蛋白转录中.
结论:
- 在外子基质中的RNA二次结构可能会推动前进方向的拼接.
- 拼接的可逆性对理解内部流动性和转移机制有影响.
- I 组内置转换可能涉及到集成到不同的RNA目标.
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