在体内和体外,通过一种不寻常的RNA元素激活c-src神经元特异拼接
1Whitehead Institute for Biomedical Research, Nine Cambridge Center, Massachusetts 02142.
Cell
|May 29, 1992
概括
一个保存的RNA序列控制了小鼠c-src N1外基的神经元特异拼接. 这种激活器序列对于神经元细胞中正确的基因表达至关重要.
科学领域:
- 分子生物学分子生物学
- 在RNA分离过程中.
- 基因规则 基因规则
背景情况:
- 小鼠的c-src基因经历了神经元特定的替代拼接.
- 了解替代拼接的调节机制对于细胞类型特定的基因表达至关重要.
研究的目的:
- 识别和描述参与小鼠c-src N1外因子神经元特异拼接的调节序列.
- 为了研究一个保存的积极作用的RNA序列在调节SRC拼接模式中的作用.
主要方法:
- 在体外剪接测试使用HeLa和Weri-1细胞提取物.
- RNA操纵以改变内子长度并评估拼接依赖性.
- 使用含有激活序列的合成RNA进行抑制试验.
主要成果:
- 一个保存的积极作用的RNA序列,位于N1外因子捐赠部位附近的内子中,对于N1外因子的神经元特异拼接至关重要.
- 缩短下游的内子赋予了构成性外子4拼接对这种激活器序列的依赖.
- 在体外复制表明,HeLa细胞跳过N1外因子,而Weri-1细胞包括它,两种模式都取决于激活剂.
- 激活器序列的合成RNA片段抑制了这两种拼接模式,这表明绑定因素的参与.
结论:
- 已识别的RNA序列作为神经元特异性c-src N1外因子含入的关键调节器.
- 该机制涉及蛋白质因子与激活器RNA的特定序列结合.
- 这种调节元件在产生c-src基因的神经元转录多样性方面发挥着关键作用.
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