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

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 端结构域是真核生物中独特的结构，可协调各种 通过翻译后修饰将转录因子转化为 Pol II。简单的共识序列 CTD (Y1S2P3T4S5P6S7) 连续重复多次并且大部分是保守的。磷酸化 序列中的两个 SP 基序已被确定为每轮转录所必需的，但 其他三个残基 Y1、T4 和 S7 在转录过程中也具有功能意义。要了解 CTD的这些翻译后修饰对转录调节的分子机制，我们结合 一种结合生物化学和生物物理研究的新型质谱方法，用于研究身份如何 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。然后我们将检查改变 CTD 修改模式的后果 转录，这将使我们能够了解调节转录的基本机制。

项目成果

Electrophoretic Mobility Shift Assay of in vitro Phosphorylated RNA Polymerase II Carboxyl-terminal Domain Substrates.

体外磷酸化 RNA 聚合酶 II 羧基末端结构域底物的电泳迁移率变化测定。

• 发表时间: 2020-06-20
• 影响因子: 0.8
• 作者: Mayfield, Joshua E;Irani, Seema;Zhang, Yan
• 通讯作者: Zhang, Yan

Simplicity is the Ultimate Sophistication-Crosstalk of Post-translational Modifications on the RNA Polymerase II.

简单性是最终的复杂性——RNA 聚合酶 II 翻译后修饰的串扰。

• 发表时间: 2021-07-09
• 影响因子: 5.6
• 作者: Venkat Ramani, Mukesh Kumar;Yang, Wanjie;Irani, Seema;Zhang, Yan
• 通讯作者: Zhang, Yan

Enzyme-mediated depletion of serum l-Met abrogates prostate cancer growth via multiple mechanisms without evidence of systemic toxicity.

酶介导的血清 L-Met 消耗可通过多种机制消除前列腺癌的生长，且没有全身毒性的证据。

• 发表时间: 2020-06-09
• 影响因子: 11.1
• 作者: Lu, Wei;Saha, Achinto;Yan, Wupeng;Garrison, Kendra;Lamb, Candice;Pandey, Renu;Irani, Seema;Lodi, Alessia;Lu, Xiyuan;Tiziani, Stefano;Zhang, Yan Jessie;Georgiou, George;DiGiovanni, John;Stone, Everett
• 通讯作者: Stone, Everett

Crystal Structure of the Ergothioneine Sulfoxide Synthase from Candidatus Chloracidobacterium thermophilum and Structure-Guided Engineering To Modulate Its Substrate Selectivity.

嗜热氯酸杆菌麦角硫因亚硫酸合酶的晶体结构和结构引导工程调节其底物选择性。

• DOI: 10.1021/acscatal.9b02054
• 发表时间: 2019-07-02
• 期刊: ACS catalysis
• 影响因子: 12.9
• 作者: Nathchar Naowarojna;S. Irani;Ronghai Cheng;W. Hu;L. Zhang;X. Li;Jia;Y. Zhang;Pinghua Liu
• 通讯作者: Pinghua Liu

Structural determinants for accurate dephosphorylation of RNA polymerase II by its cognate C-terminal domain (CTD) phosphatase during eukaryotic transcription.

在真核转录过程中，RNA 聚合酶 II 的同源 C 末端结构域 (CTD) 磷酸酶准确去磷酸化的结构决定因素。

• 发表时间: 2019
• 作者: Irani, Seema;Sipe, Sarah N;Yang, Wanjie;Burkholder, Nathaniel T;Lin, Brian;Sim, Kelly;Matthews, Wendy L;Brodbelt, Jennifer S;Zhang, Yan
• 通讯作者: Zhang, Yan