Identification of novel RNA Polymerase II CTD interaction sites on the mRNA Capping Enzyme
| Autor: | Andrei V. Pisliakov, Rajaei Almohammed, Marcus G Bage, Victoria H. Cowling |
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| Rok vydání: | 2020 |
| Předmět: |
0303 health sciences
Messenger RNA 010304 chemical physics Polyadenylation biology Chemistry fungi genetic processes RNA polymerase II environment and public health 01 natural sciences Cell biology enzymes and coenzymes (carbohydrates) 03 medical and health sciences Transcription (biology) 0103 physical sciences RNA splicing health occupations biology.protein Phosphorylation CTD Nuclear export signal 030304 developmental biology |
| DOI: | 10.1101/2020.02.25.964700 |
| Popis: | 1AbstractRecruitment of the mRNA Capping Enzyme (CE/RNGTT) to the site of transcription is essential for the formation of the 5’ mRNA cap, which in turn ensures efficient transcription, splicing, polyadenylation, nuclear export and translation of mRNA in eukaryotic cells. The CE is recruited and activated by the Serine-5 phosphorylated carboxyl-terminal domain (CTD) of RNA polymerase II. Through the use of molecular dynamics simulations and enhanced sampling techniques, we provide a systematic and detailed characterisation of the human CE-CTD interface, describing the effect of the CTD phosphorylation state, length and orientation on this interaction. Our computational analyses identify novel CTD interaction sites on the human CE surface and quantify their relative contributions to CTD binding. We also identify differences in the CTD binding conformation when phosphorylated at either the Serine-2 or Serine-5 positions, thus providing insights into how the CE reads the CTD code. The computational findings are then validated by binding and activity assays. These novel CTD interaction sites are compared with cocrystal structures of the CE-CTD complex in different eukaryotic taxa, leading to the conclusion that this interface is considerably more conserved than previous structures have indicated. |
| Databáze: | OpenAIRE |
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