Structural Motifs for CTD Kinase Specificity on RNA Polymerase II during Eukaryotic Transcription
| Autor: | Edwin E. Escobar, Jamie P. Butalewicz, Haoyi Wu, Seema Irani, Joshua E. Mayfield, Meena Tadros, Jennifer S. Brodbelt, Yan Jessie Zhang, Mukesh Kumar Venkat Ramani, Victoria C. Cotham, Rosamaria Y. Moreno |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Transcription Genetic viruses RNA polymerase II 01 natural sciences Biochemistry environment and public health Article Mass Spectrometry Substrate Specificity Serine 03 medical and health sciences Discoidin Domain Receptor 1 Transcription (biology) Humans Amino Acid Sequence Phosphorylation Structural motif biology Sequence Homology Amino Acid 010405 organic chemistry Kinase Chemistry Eukaryotic transcription General Medicine Cyclin-Dependent Kinases 0104 chemical sciences Cell biology enzymes and coenzymes (carbohydrates) 030104 developmental biology biology.protein Molecular Medicine CTD RNA Polymerase II Protein Kinases Protein Processing Post-Translational Cyclin-Dependent Kinase-Activating Kinase |
| Zdroj: | ACS Chem Biol |
| Popis: | The phosphorylation states of RNA polymerase II coordinate the process of eukaryotic transcription by recruitment of transcription regulators. The individual residues of the repetitive heptad of the C-terminal domain (CTD) of the biggest subunit of RNA polymerase II are phosphorylated temporally at different stages of transcription. Intriguingly, despite similar flanking residues, phosphorylation of Ser2 and Ser5 in CTD heptads play dramatically different roles. The mechanism of how the kinases place phosphorylation on the correct serine is not well understood. In this paper, we use biochemical assays, mass spectrometry, molecular modeling, and structural analysis to understand the structural elements determining which serine of the CTD heptad is subject to phosphorylation. We identified three motifs in the activation/P+1 loops differentiating the intrinsic specificity of CTD in various CTD kinases. We characterized the enzyme specificity of the CTD kinases-CDK7 as Ser5-specific, Erk2 with dual specificity for Ser2 and Ser5, and Dyrk1a as a Ser2-specific kinase. We also show that the specificities of kinases are malleable and can be modified by incorporating mutations in their activation/P+1 loops that alter the interactions of the three motifs. Our results provide an important clue to the understanding of post-translational modification of RNA polymerase II temporally during active transcription. |
| Databáze: | OpenAIRE |
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