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| Status |
Public on Jun 16, 2016 |
| Title |
Pol II and CTD Phosphorylation ChIP-chip of ATP-analog sensitive Kin28 budding yeast |
| Platform organism |
Saccharomyces cerevisiae |
| Sample organisms |
Saccharomyces cerevisiae; Saccharomyces cerevisiae BY4741 |
| Experiment type |
Genome binding/occupancy profiling by genome tiling array
|
| Summary |
During transcription initiation, the TFIIH-kinase Kin28/Cdk7 marks RNA polymerase II (Pol II) by phosphorylating the C-terminal domain (CTD) of its largest subunit. Here we describe a structure-guided chemical approach to covalently and specifically inactivate Kin28 kinase activity in vivo. This method of irreversible inactivation recapitulates both the lethal phenotype and the key molecular signatures that result from genetically disrupting Kin28 function in vivo. Inactivating Kin28 impacts promoter release to differing degrees and reveals a “checkpoint” during the transition to productive elongation. While promoter-proximal pausing is not observed in budding yeast, inhibition of Kin28 attenuates elongation-licensing signals, resulting in Pol II accumulation at the +2 nucleosome and reduced transition to productive elongation. Furthermore, upon inhibition, global stabilization of mRNA masks different degrees of reduction in nascent transcription. This study resolves long-standing controversies on the role of Kin28 in transcription and provides a rational approach to irreversibly inhibit other kinases in vivo.
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| Overall design |
Rpb3-TAP, Ser2P, Ser5P, and Ser7P ChIP of irreversibly sensitive kin28is strain treated with inhibitors 1-NAPP-1 and CMK or solvent control (DMSO)
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| Contributor(s) |
Ansari AZ, Rodríguez-Molina JB |
| Citation(s) |
27477907 |
| |
| Submission date |
Jun 15, 2016 |
| Last update date |
Sep 20, 2016 |
| Contact name |
Aseem Ansari |
| Organization name |
University of Wisconsin-Madison
|
| Department |
Biochemistry and The Genome Center
|
| Lab |
Ansari
|
| Street address |
440 Henry Mall
|
| City |
Madison |
| State/province |
WI |
| ZIP/Postal code |
53706 |
| Country |
USA |
| |
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| Platforms (2) |
| GPL22008 |
S. cerevisiae 120807_SCer_Dwolbach_CGH |
| GPL22009 |
S. cerevisiae 2006-10-12_Ansari_tiling_51mer |
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| Samples (20)
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| This SubSeries is part of SuperSeries: |
| GSE83417 |
Pol II and CTD Phosphorylation of ATP-analog sensitive Kin28 budding yeast |
|
| Relations |
| BioProject |
PRJNA325831 |
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