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.

抽象的 最大的RNA聚合酶II亚基的C末端结构域（CTD）是独特的CTD序列 （Y1S2P3T4S5P6S7）重复多次，并且在真核生物中大多是保守的。该域协调 通过翻译后修改将转录因子招募到Pol II，其损失 削弱高效的转录过程，并导致细胞死亡。准确的磷酸化状态 激酶和磷酸酶在CTD七含量重复序列中的不同残留物中对于精确募集至关重要 蛋白质介导转录过程。我们的实验室利用我们在蛋白质化学方面的丰富经验 通过研究活性和特异性，了解转录过程中磷酸化的精确模式 这些激酶和磷酸酶。我们试图了解CTD的翻译后修改 在生物事件期间发生了变化，以及如何反映在转录结果中。我们使用 多学科方法，包括结构生物学，生物化学，质谱法和 全局转录组分析研究了RNA不同修饰状态的分子机制 聚合酶II协调真核转录。 配备了有关磷酸酶功能和活性的广泛知识，我们进一步探讨了如何 SCP1的化学抑制作用，一种属于超家族的人磷酸酶，阻碍了一些生长 胶质母细胞瘤细胞。我们利用基于结构的抑制剂设计的经验来识别共价和非 - 靶向这种独特的磷酸酶的共价抑制剂。