Proteomics

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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]

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