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Related Experiment Video

Updated: Jan 5, 2026

Identification of Cyclin-dependent Kinase 1 Specific Phosphorylation Sites by an In Vitro Kinase Assay
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CDK12 Activity-Dependent Phosphorylation Events in Human Cells.

Bartlomiej Bartkowiak1, Christopher M Yan2, Erik J Soderblom3

  • 1Department of Biochemistry, Duke Med. Ctr., Durham, NC 27710, USA. bartjbartkowiak@gmail.com.

Biomolecules
|October 27, 2019
PubMed
Summary
This summary is machine-generated.

The cyclin-dependent kinase 12 (CDK12) enzyme phosphorylates more than just the RPB1 CTD. This study identified novel CDK12 substrates involved in transcription, RNA processing, and other cellular events.

Keywords:
1-NM-PP1CTDRNA polymerase IIRNA processingTPRXPCanalog-sensitive CTD kinasemESC undifferentiated statemRNA nuclear exportnuclear poretranscription

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Cell Biology

Background:

  • Cyclin-dependent kinase 12 (CDK12/CyclinK) is known to phosphorylate the C-terminal domain (CTD) of RPB1.
  • Its broader substrate specificity and cellular functions remain incompletely understood.

Purpose of the Study:

  • To investigate whether CDK12 phosphorylates substrates beyond the RPB1 CTD.
  • To identify novel CDK12 activity-dependent phosphorylation events in human cells.
  • To explore the roles of CDK12 in various cellular processes.

Main Methods:

  • Utilized a CDK12 analog-sensitive HeLa cell line.
  • Performed phospho-proteomic analysis before and after selective CDK12 inhibition using 1-NM-PP1.
  • Quantified changes in phosphopeptide abundance.

Main Results:

  • Identified 5,644 distinct phosphopeptides; 50 showed >2-fold decrease upon CDK12 inhibition.
  • Affected phosphopeptides were not derived from RPB1.
  • Identified proteins involved in transcription, RNA processing, mRNA nuclear export, cell differentiation, and mitosis.

Conclusions:

  • CDK12 regulates phosphorylation events beyond the RPB1 CTD.
  • CDK12 plays roles in diverse cellular processes including transcription, RNA processing, mRNA export, differentiation, and mitosis.
  • Some novel CDK12 substrates are direct targets, while others may be indirectly affected.