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Analog-sensitive cell line identifies cellular substrates of CDK9.
Oncotarget 10, 6934-6943 (2019)
Transcriptional cyclin-dependent kinases regulate all phases of transcription. Cyclin-dependent kinase 9 (CDK9) has been implicated in the regulation of promoter-proximal pausing of RNA polymerase II and more recently in transcription termination. Study of the substrates of CDK9 has mostly been limited to in vitro approaches that lack a quantitative assessment of CDK9 activity. Here we analyzed the cellular phosphoproteome upon inhibition of CDK9 by combining analog-sensitive kinase technology with quantitative phosphoproteomics in Raji B-cells. Our analysis revealed the activity of CDK9 on 1102 phosphosites quantitatively, and we identified 120 potential cellular substrates. Furthermore, a substantial number of CDK9 substrates were described as splicing factors, highlighting the role of CDK9 in transcription-coupled splicing events. Based on comparison to in vitro data, our findings suggest that cellular context fundamentally impacts the activity of CDK9 and specific selection of its substrates.
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Publication type Article: Journal article
Document type Scientific Article
Keywords Cdk9 ; Rna Splicing ; Phosphoproteomics ; Protein Kinase ; Transcription
ISSN (print) / ISBN 1949-2553
Quellenangaben Volume: 10, Issue: 65, Pages: 6934-6943
Publisher Impact Journals LLC
Institute(s) Institute of Functional Epigenetics (IFE)