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Identification of cellular CDK9 substrates using an analog-sensitive cell line

ABSTRACT: Analog-sensitive kinase technology allows for specific inhibition of a given kinase. Here we utilize an analog-sensitive CDK9 cell line to determine the impact of CDK9 inhibition on the phospho-proteome of human cells. Specifically, we aim to identify substrates of CDK9 in a quantitative way using stable isotope labeling by amino acids in cell culture (SILAC) followed by LC-MS/MS.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): B Cell

SUBMITTER: Ignasi Forne

LAB HEAD: Prof. Dr. Dirk Eick

PROVIDER: PXD014825 | Pride | 2019-11-11

REPOSITORIES: Pride

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Analog-sensitive cell line identifies cellular substrates of CDK9.

Decker Tim-Michael TM Forné Ignasi I Straub Tobias T Elsaman Hesham H Ma Guoli G Shah Nilay N Imhof Axel A Eick Dirk D

Oncotarget 20191210 65

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 <i>in vitro</i> approaches that lack a quantitative assessment of CDK9 activity. Here we analyzed the cellular phosphoproteome upon inhibition of CDK9 by combining analog-sensitive kinase techn ...[more]

PMID: 31857848

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