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: 10158496
• 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 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10158496
• 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; advanced analytics; biophysical analysis; combinatorial; experimental study; in vivo; innovation; inorganic phosphate; novel; reconstruction; recruit; tandem mass spectrometry; transcription factor; ultraviolet; 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; advanced analytics; 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.

抽象的 RNA聚合酶II（CTD）的C末端结构域是真核生物中的独特结构，可以协调各种 通过其翻译后修饰向POL II转录因素。简单的共识序列 CTD（Y1S2P3T4S5P6S7）连续多次重复，并且大多是保守的。磷酸化 序列中的两个SP基序已被确定为每一轮转录至关重要，但是 在转录过程中，其他三个残基Y1，T4和S7也具有功能含义。理解 CTD在转录调制时的这些翻译后修饰的分子机制，我们结合了 一种新型的质谱法，并具有生化和生物物理研究，以研究身份如何 这三个位置的修改状态影响RNA的翻译后修饰状态 聚合酶II。然后，我们将检查改变CTD修改模式的后果 转录，这将使我们能够理解调节转录的基本机制。