What's all the phos about? Insights into the phosphorylation state of the RNA polymerase II C-terminal domain via mass spectrometry,RSC Chemical Biology

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What's all the phos about? Insights into the phosphorylation state of the RNA polymerase II C-terminal domain via mass spectrometry
RSC Chemical Biology ( IF 4.2 ) Pub Date : 2021-6-3 , DOI: 10.1039/d1cb00083g
Blase M LeBlanc ₁ , R Yvette Moreno ₁ , Edwin E Escobar ₂ , Mukesh Kumar Venkat Ramani ₁ , Jennifer S Brodbelt ₂ , Yan Zhang _{1,

3}

Affiliation

RNA polymerase II (RNAP II) is one of the primary enzymes responsible for expressing protein-encoding genes and some small nuclear RNAs. The enigmatic carboxy-terminal domain (CTD) of RNAP II and its phosphorylation state are critically important in regulating transcription in vivo. Early methods of identifying phosphorylation on the CTD heptad were plagued by issues of low specificity and ambiguous signals. However, advancements in the field of mass spectrometry (MS) have presented the opportunity to gain new insights into well-studied processes as well as explore new frontiers in transcription. By using MS, residues which are modified within the CTD heptad and across repeats are now able to be pinpointed. Likewise, identification of kinase and phosphatase specificity towards residues of the CTD has reached a new level of accuracy. Now, MS is being used to investigate the crosstalk between modified residues of the CTD and may be a critical technique for understanding how phosphorylation plays a role in the new LLPS model of transcription. Herein, we discuss the development of various MS techniques and evaluate their capabilities. By highlighting the pros and cons of each technique, we aim to provide future investigators with a comprehensive overview of how MS can be used to investigate the complexities of RNAP-II mediated transcription.

中文翻译：

所有的 phos 是关于什么的？通过质谱分析 RNA 聚合酶 II C 末端结构域的磷酸化状态

RNA 聚合酶 II (RNAP II) 是负责表达蛋白质编码基因和一些小核 RNA 的主要酶之一。RNAP II 的神秘羧基末端结构域 (CTD) 及其磷酸化状态对于调节体内转录至关重要。早期鉴定 CTD 七肽磷酸化的方法受到特异性低和信号模糊问题的困扰。然而，质谱 (MS) 领域的进步提供了对经过充分研究的过程获得新见解以及探索转录新领域的机会。通过使用 MS，现在能够精确定位 CTD 七肽内和重复序列中修饰的残基。同样，对 CTD 残基的激酶和磷酸酶特异性的鉴定也达到了新的准确度水平。现在，MS 被用来研究 CTD 修饰残基之间的串扰，并且可能是了解磷酸化如何在新的 LLPS 转录模型中发挥作用的关键技术。在这里，我们讨论了各种 MS 技术的发展并评估了它们的能力。通过强调每种技术的优缺点，我们的目标是为未来的研究人员提供如何使用 MS 来研究 RNAP-II 介导的转录的复杂性的全面概述。

更新日期：2021-06-08

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