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The structure and substrate specificity of human Cdk12/Cyclin K

Phosphorylation of the RNA polymerase II C-terminal domain (CTD) by cyclin-dependent kinases is important for productive transcription. Here we determine the crystal structure of Cdk12/CycK and analyse its requirements for substrate recognition. Active Cdk12/CycK is arranged in an open conformation...

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Autores principales: Bösken, Christian A., Farnung, Lucas, Hintermair, Corinna, Merzel Schachter, Miriam, Vogel-Bachmayr, Karin, Blazek, Dalibor, Anand, Kanchan, Fisher, Robert P., Eick, Dirk, Geyer, Matthias
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Pub. Group 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3973122/
https://www.ncbi.nlm.nih.gov/pubmed/24662513
http://dx.doi.org/10.1038/ncomms4505
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author Bösken, Christian A.
Farnung, Lucas
Hintermair, Corinna
Merzel Schachter, Miriam
Vogel-Bachmayr, Karin
Blazek, Dalibor
Anand, Kanchan
Fisher, Robert P.
Eick, Dirk
Geyer, Matthias
author_facet Bösken, Christian A.
Farnung, Lucas
Hintermair, Corinna
Merzel Schachter, Miriam
Vogel-Bachmayr, Karin
Blazek, Dalibor
Anand, Kanchan
Fisher, Robert P.
Eick, Dirk
Geyer, Matthias
author_sort Bösken, Christian A.
collection PubMed
description Phosphorylation of the RNA polymerase II C-terminal domain (CTD) by cyclin-dependent kinases is important for productive transcription. Here we determine the crystal structure of Cdk12/CycK and analyse its requirements for substrate recognition. Active Cdk12/CycK is arranged in an open conformation similar to that of Cdk9/CycT but different from those of cell cycle kinases. Cdk12 contains a C-terminal extension that folds onto the N- and C-terminal lobes thereby contacting the ATP ribose. The interaction is mediated by an HE motif followed by a polybasic cluster that is conserved in transcriptional CDKs. Cdk12/CycK showed the highest activity on a CTD substrate prephosphorylated at position Ser7, whereas the common Lys7 substitution was not recognized. Flavopiridol is most potent towards Cdk12 but was still 10-fold more potent towards Cdk9. T-loop phosphorylation of Cdk12 required coexpression with a Cdk-activating kinase. These results suggest the regulation of Pol II elongation by a relay of transcriptionally active CTD kinases.
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spelling pubmed-39731222014-04-03 The structure and substrate specificity of human Cdk12/Cyclin K Bösken, Christian A. Farnung, Lucas Hintermair, Corinna Merzel Schachter, Miriam Vogel-Bachmayr, Karin Blazek, Dalibor Anand, Kanchan Fisher, Robert P. Eick, Dirk Geyer, Matthias Nat Commun Article Phosphorylation of the RNA polymerase II C-terminal domain (CTD) by cyclin-dependent kinases is important for productive transcription. Here we determine the crystal structure of Cdk12/CycK and analyse its requirements for substrate recognition. Active Cdk12/CycK is arranged in an open conformation similar to that of Cdk9/CycT but different from those of cell cycle kinases. Cdk12 contains a C-terminal extension that folds onto the N- and C-terminal lobes thereby contacting the ATP ribose. The interaction is mediated by an HE motif followed by a polybasic cluster that is conserved in transcriptional CDKs. Cdk12/CycK showed the highest activity on a CTD substrate prephosphorylated at position Ser7, whereas the common Lys7 substitution was not recognized. Flavopiridol is most potent towards Cdk12 but was still 10-fold more potent towards Cdk9. T-loop phosphorylation of Cdk12 required coexpression with a Cdk-activating kinase. These results suggest the regulation of Pol II elongation by a relay of transcriptionally active CTD kinases. Nature Pub. Group 2014-03-24 /pmc/articles/PMC3973122/ /pubmed/24662513 http://dx.doi.org/10.1038/ncomms4505 Text en Copyright © 2014, Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved. http://creativecommons.org/licenses/by-nc-by/3.0/ This work is licensed under a Creative Commons Attribution 3.0 Unported License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/3.0/
spellingShingle Article
Bösken, Christian A.
Farnung, Lucas
Hintermair, Corinna
Merzel Schachter, Miriam
Vogel-Bachmayr, Karin
Blazek, Dalibor
Anand, Kanchan
Fisher, Robert P.
Eick, Dirk
Geyer, Matthias
The structure and substrate specificity of human Cdk12/Cyclin K
title The structure and substrate specificity of human Cdk12/Cyclin K
title_full The structure and substrate specificity of human Cdk12/Cyclin K
title_fullStr The structure and substrate specificity of human Cdk12/Cyclin K
title_full_unstemmed The structure and substrate specificity of human Cdk12/Cyclin K
title_short The structure and substrate specificity of human Cdk12/Cyclin K
title_sort structure and substrate specificity of human cdk12/cyclin k
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3973122/
https://www.ncbi.nlm.nih.gov/pubmed/24662513
http://dx.doi.org/10.1038/ncomms4505
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