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

真核转录过程中RNA聚合酶II磷酸化密码的解读

• 批准号: 9751900
• 负责人: Jennifer S. Brodbelt
• 金额: $ 34.7万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9751900
• 关键词: Address; Affect; Amino Acids; Binding; Binding Proteins; Biochemical; Biological; Biological Models; Biology; C-terminal; Cell Death; Chemicals; Code; Consensus; Consensus Sequence; DNA Polymerase II; Defect; Development; Enzymes; Eukaryota; Evaluation; Event; Generations; Genetic Transcription; Gold; In Vitro; Lead; Length; Malignant Neoplasms; Mass Spectrum Analysis; Messenger RNA; Methodology; Methods; Modeling; Modification; Molecular; Organism; Outcome; Pattern; Phenotype; Phosphorylation; Phosphorylation Site; Photons; Positioning Attribute; Post-Translational Modification Site; Post-Translational Protein Processing; Process; Proteins; RNA Polymerase II; Resolution; Role; Saccharomyces cerevisiae; Site; Small Nuclear RNA; Structure; Techniques; Testing; Time; Transcription Initiation; Transcription Process; Transcription Regulatory Protein; Transcriptional Regulation; biophysical analysis; combinatorial; experimental study; in vivo; innovation; inorganic phosphate; novel; reconstruction; recruit; tandem mass spectrometry; transcription factor; ultraviolet

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.

摘要 RNA聚合酶II（CTD）的C末端结构域是真核生物中的独特结构，其协调各种RNA聚合酶II（CTD）。 Pol II通过其翻译后修饰与转录因子结合。简单的共有序列 CTD（Y1S2P3T4S5P6S7）连续重复多次，并且大部分是保守的。的磷酸化 序列中的两个SP基序已被鉴定为对每轮转录都是必需的，但 其他三个残基Y1、T4和S7在转录过程中也具有功能性意义。了解 CTD的这些翻译后修饰对转录调节的分子机制，我们联合收割机 一种新的质谱分析方法与生物化学和生物物理研究，以调查如何身份 这三个位置的修饰状态影响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，这将使我们能够理解这种调节转录的基本机制。