通过CDK9依赖的SPT5的三方酸化来协调RNA聚合酶II转录周期的控制
bioRxiv : the preprint server for biology
|August 30, 2024
概括
通过CDK9对SPT5蛋白的酸化调节了转录. 这种三方酸化控制RNA聚合酶II的暂停,延长和RNA处理,确保细胞活力.
科学领域:
- 分子生物学分子生物学
- 基因表达规范 基因表达规范
- 生物化学 生物化学
背景情况:
- RNA聚合酶II (RNAPII) 的转录周期是通过可逆蛋白酸化动态调节的.
- 延长因子SPT5具有关键的酸化位点,这些位点是循环林依赖激酶9 (CDK9) 的目标.
- 这些位点位于KOWx-4/5链接器,碳氧终端重复1 (CTR1) 和碳氧终端重复2 (CTR2) 中.
研究的目的:
- 研究SPT5KOWx-4/5链接器,CTR1和CTR2酸化在调节转录中的协调作用.
- 确定这些酸化事件如何影响RNAPII暂停,延长速度和RNA处理.
- 阐明SPT5酸化对细胞增殖和活力的影响.
主要方法:
- 在特定的SPT5区域预防或模仿酸化的局部定向突变发生 (KOWx-4/5链接器,CTR1,CTR2).
- 使用生物化学分析分析RNAPII暂停,延长率和新生的转录.
- 评估RNA处理,包括拼接和终止,以及稳定状态mRNA水平.
- 在SPT5突变后评估细胞增殖和活力.
主要成果:
- 阻止CTR1或CTR2酸化的突变增加了暂停,而突变两种CTR减弱了这种效应.
- 单个CTR突变对延长速度和转录有不同的影响,在组合CTR突变时观察到部分逆转.
- 两种CTR同时发生突变导致异常拼接,失调终结,mRNA水平降低,细胞增殖严重受损.
结论:
- 三方SPT5酸化作为一个关键的调节器,协调暂停释放和调节RNAPII延长率.
- 这种酸化确保了高效准确的转录,包括RNA处理和终结.
- 不调节SPT5酸化显著影响细胞活力,突出其在维持细胞功能的重要作用.
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