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CDK12 controls G1/S progression by regulating RNAPII processivity at core DNA replication genes.

Anil Paul Chirackal Manavalan1, Kveta Pilarova1, Michael Kluge2

  • 1Central European Institute of Technology (CEITEC), Masaryk University, Brno, Czech Republic.

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|July 27, 2019
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Summary

Cyclin-dependent kinase 12 (CDK12) activity is crucial for DNA replication gene transcription and cell cycle progression. CDK12 inhibition causes RNA polymerase II defects, impacting DNA repair and genome stability in cancer.

Keywords:
CDK12CTD Ser2 phosphorylationG1/Spremature termination and polyadenylationtandem duplications

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Area of Science:

  • Molecular Biology
  • Cancer Biology
  • Genetics

Background:

  • Cyclin-dependent kinase 12 (CDK12) is implicated in RNA polymerase II (RNAPII) elongation and is frequently altered in cancers.
  • Previous studies suggest CDK12 depletion affects homologous recombination (HR) DNA repair gene expression, but its broader roles remain unclear.

Purpose of the Study:

  • To investigate the function of CDK12 kinase activity in gene transcription and cellular processes using a chemical genetic approach.
  • To elucidate the impact of CDK12 inhibition on RNAPII dynamics and identify its target genes and pathways.

Main Methods:

  • Utilized a chemical genetic strategy with an analog-sensitive CDK12 inhibitor.
  • Performed RNA sequencing (RNA-seq) and chromatin immunoprecipitation sequencing (ChIP-seq) to analyze gene expression and RNAPII occupancy.
  • Assessed RNAPII processivity and phosphorylation status (RNAPII-Ser2).

Main Results:

  • CDK12 kinase activity is essential for the transcription of core DNA replication genes, driving G1/S phase progression.
  • CDK12 inhibition leads to RNAPII processivity defects, marked by reduced signal at the 3' ends of long, poly(A)-rich genes.
  • While global RNAPII-Ser2 phosphorylation is unaffected, specific CDK12-dependent genes show altered P-Ser2 peak localization correlating with transcription loss.

Conclusions:

  • CDK12 catalytic activity provides a novel link between transcriptional regulation and cell cycle control.
  • CDK12 inactivation may contribute to cancer genome instability through impaired DNA replication and HR DNA repair pathways.