title: The structure and substrate specificity of human Cdk12/Cyclin K.
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.
authors.affiliation: 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.
authors.forename: Christian A
authors.initials: CA
authors.lastname: Bösken
authors.affiliation: Department of Physical Biochemistry, Max Planck Institute of Molecular Physiology, Otto-Hahn-Strasse 11, Dortmund 44227, Germany.
authors.forename: Lucas
authors.initials: L
authors.lastname: Farnung
authors.affiliation: Department of Molecular Epigenetics, Helmholtz Center Munich, Center for Integrated Protein Science (CIPSM), Marchioninistrasse 25, München 81377, Germany.
authors.forename: Corinna
authors.initials: C
authors.lastname: Hintermair
authors.affiliation: Department of Structural and Chemical Biology, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
authors.forename: Miriam
authors.initials: M
authors.lastname: Merzel Schachter
authors.affiliation: Department of Physical Biochemistry, Max Planck Institute of Molecular Physiology, Otto-Hahn-Strasse 11, Dortmund 44227, Germany.
authors.forename: Karin
authors.initials: K
authors.lastname: Vogel-Bachmayr
authors.affiliation: Central European Institute of Technology (CEITEC), Masaryk University, Brno 62500, Czech Republic.
authors.forename: Dalibor
authors.initials: D
authors.lastname: Blazek
authors.affiliation: Group Physical Biochemistry, Center of Advanced European Studies and Research, Ludwig-Erhard-Allee 2, Bonn 53175, Germany.
authors.forename: Kanchan
authors.initials: K
authors.lastname: Anand
authors.affiliation: Department of Structural and Chemical Biology, Icahn School of Medicine at Mount Sinai, New York, New York 10029, USA.
authors.forename: Robert P
authors.initials: RP
authors.lastname: Fisher
authors.affiliation: Department of Molecular Epigenetics, Helmholtz Center Munich, Center for Integrated Protein Science (CIPSM), Marchioninistrasse 25, München 81377, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Eick
authors.affiliation: 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.
authors.forename: Matthias
authors.initials: M
authors.lastname: Geyer
mesh_terms: D015153:Blotting, Western; D003460:Crystallization; D018844:Cyclin-Dependent Kinases / Q000737:chemistry* / Q000378:metabolism; D016213:Cyclins / Q000737:chemistry* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D008958:Models, Molecular; D046912:Multiprotein Complexes / Q000737:chemistry* / Q000378:metabolism; D011487:Protein Conformation; D013379:Substrate Specificity
chemical_list: C113028:CCNK protein, human; D016213:Cyclins; D046912:Multiprotein Complexes; C447339:CDK12 protein, human; D018844:Cyclin-Dependent Kinases