Deciphering the phosphorylation pattern of RNA polymerase II for eukaryotic transcription

破译真核转录中 RNA 聚合酶 II 的磷酸化模式

• 批准号: 10552217
• 负责人: Yan Jessie Zhang
• 金额: $ 48.92万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-06 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10552217
• 关键词: Biochemistry; Biological; Biology; C-terminal; Cell Death; Cells; Chemicals; DNA Polymerase II; Defect; Development; Eukaryota; Event; Genetic Transcription; Glioblastoma; Growth; Human; Knowledge; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Methodology; Methods; Modification; Molecular; Outcome; Pattern; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Post-Translational Protein Processing; Process; Protein Chemistry; Proteins; RNA Polymerase II; RNA polymerase II largest subunit; Research; Specificity; Transcription Process; design; experience; inhibitor; multidisciplinary; recruit; structural biology; transcription factor; transcriptome

ABSTRACT The C-terminal domain (CTD) of the largest RNA polymerase II subunit is a unique CTD sequence (Y1S2P3T4S5P6S7) repeated many times and is mostly conserved in eukaryotes. This domain coordinates the recruitment of transcriptional factors to Pol II through its post-translational modifications, the loss of which cripples the highly efficient transcription process and causes the cell to die. The accurate phosphorylation state of different residues in the CTD heptad repeats by kinase and phosphatases is crucial to precisely recruiting proteins to mediate the transcription process. Our lab utilizes our extensive experience in protein chemistry to understand the precise pattern of phosphorylation during transcription by investigating the activity and specificity of these kinases and phosphatases. We seek to understand how the post-translational modifications of CTD are altered during biological events and how such changes are reflected in the outcome of transcription. We use multi-disciplinary methods including structural biology, biochemistry, mass spectrometry methodology, and global transcriptome analysis to investigate the molecular mechanism of how different modification states of RNA polymerase II coordinate the eukaryotic transcription. Equipped with extensive knowledge about the phosphatase function and activity, we further explore how the chemical inhibition of SCP1, a human phosphatase belonging to HAD superfamily, thwarts some of the growth glioblastoma cells. We utilized our experience in structural-based inhibitor design to identify covalent and non- covalent inhibitors targeting this unique phosphatase.

摘要 最大的RNA聚合酶II亚基的C-末端结构域（CTD）是唯一的CTD序列 （Y1 S2 P3 T4 S5 P6 S7）重复多次，在真核生物中大部分保守。此域协调 通过其翻译后修饰将转录因子募集至Pol II， 削弱了高效的转录过程并导致细胞死亡。准确的磷酸化状态 通过激酶和磷酸酶对CTD七肽重复序列中的不同残基的识别对于精确招募至关重要 蛋白质来介导转录过程。我们的实验室利用我们丰富的蛋白质化学经验， 通过研究磷酸化的活性和特异性， 这些激酶和磷酸酶。我们试图了解CTD的翻译后修饰是如何在 在生物学事件中改变，以及这些变化如何反映在转录的结果中。我们使用 多学科方法，包括结构生物学、生物化学、质谱方法学， 全局转录组分析以研究RNA不同修饰状态的分子机制 聚合酶II协调真核转录。 配备了广泛的知识，磷酸酶的功能和活性，我们进一步探讨如何 SCP 1是一种属于HAD超家族的人类磷酸酶，它的化学抑制作用阻碍了某些生长 胶质瘤细胞我们利用我们在基于结构的抑制剂设计方面的经验来鉴定共价和非共价的抑制剂。 针对这种独特的磷酸酶的共价抑制剂。