Interpretation of the phosphorylation code of RNA polymerase II during eukaryotic transcription

摘要 RNA聚合酶II(CTD)的C末端结构域是真核生物中一种独特的结构，它协调各种 转录因子通过POL II的翻译后修饰。简单的共识序列 CTD(Y1S2P3T4S5P6S7)是连续多次重复的，大部分是保守的。蛋白的磷酸化 该序列中的两个SP基序已被鉴定为对每一轮转录都是必不可少的，但 另外三个残基Y1、T4和S7在转录过程中也有功能暗示。要了解 CTD的这些翻译后修饰对转录调控的分子机制，我们结合 一种结合生化和生物物理研究的新的质谱学方法来研究 这三个位置的修饰状态影响着RNA的翻译后修饰状态 聚合酶II。然后我们将研究改变CTD的修饰模式对 转录，这将使我们了解这一调节转录的基本机制。

ABSTRACT The C-terminal domain of RNA polymerase II (CTD) is a unique structure in eukaryotes that coordinates various transcriptional factors to Pol II through its post-translational modifications. The simple consensus sequence of CTD (Y1S2P3T4S5P6S7) is repeated many times consecutively and is mostly conserved. The phosphorylation of the two SP motifs in the sequence has been identified to be essential for every round of transcription but the other three residues, Y1, T4 and S7, also have functional implications during transcription. To understand the molecular mechanism of these post-translational modifications of CTD on transcription modulation, we combine a novel mass spectrometry methodology with biochemical and biophysical studies to investigate how the identity and modification states of these three positions affect the post-translational modification states of RNA polymerase II. We will then examine the consequences of altering the modification patterns of CTD on transcription, which will allow us to understand this fundamental mechanism that regulates transcription.