CDK12 catalytic activity is rate-limiting for RNAPII processivity on core DNA replication genes and G1/S progression (ChIP-Seq)

In this study we characterized the importance of CDK12-kinase activity in cell cycle regulation, using CDK12 anolog-sentive cells. Inhibition of analog-sensitive CDK12 reveals its catalytic activity is necessary for optimal G1/S progression. Mechanistically, CDK12 regulates transcription of core DNA replication genes and affects timely assembly of pre-replication DNA complex on chromatin. We have performed 3'-end RNA-sequencing after CDK12 inhibition and identified that the expression of core DNA replication genes were affected. To investigate further, we carried out nuclear RNA-seq coupled with ChIP-seq, and demonstrated that CDK12 regulates RNAPII processivity of core DNA replication genes and optimal G1/S progression. Overall design: After treatment with or without 5 µM 3-MB-PP1 for 4.5 h in serum synchronised AS CDK12 HCT116 cells, ChIP-seq was performed in three biological replicates with RNAPII, P-Ser2 RNAPII and P-Ser5 RNAPII antibodies. Total number of samples analyzed were twenty, including two no antibody control samples