The structure and substrate specificity of human Cdk12/Cyclin K

  • Nat Commun. 2014 Mar 24;5:3505. doi: 10.1038/ncomms4505.
Christian A Bösken  1 ,  Lucas Farnung  2 ,  Corinna Hintermair  3 ,  Miriam Merzel Schachter  4 ,  Karin Vogel-Bachmayr  2 ,  Dalibor Blazek  5 ,  Kanchan Anand  6 ,  Robert P Fisher  4 ,  Dirk Eick  3 ,  Matthias Geyer  1
Affiliations
  • 1. 1] Group Physical Biochemistry, Center of Advanced European Studies and Research, Ludwig-Erhard-Allee 2, Bonn 53175, Germany [2] Department of Physical Biochemistry, Max Planck Institute of Molecular Physiology, Otto-Hahn-Strasse 11, Dortmund 44227, Germany.
  • 2. Department of Physical Biochemistry, Max Planck Institute of Molecular Physiology, Otto-Hahn-Strasse 11, Dortmund 44227, Germany.
  • 3. Department of Molecular Epigenetics, Helmholtz Center Munich, Center for Integrated Protein Science (CIPSM), Marchioninistrasse 25, München 81377, Germany.
  • 4. Department of Structural and Chemical Biology, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
  • 5. Central European Institute of Technology (CEITEC), Masaryk University, Brno 62500, Czech Republic.
  • 6. Group Physical Biochemistry, Center of Advanced European Studies and Research, Ludwig-Erhard-Allee 2, Bonn 53175, Germany.
PMID: 24662513 DOI: 10.1038/ncomms4505
Abstract

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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