|
| Status |
Public on Jun 07, 2018 |
| Title |
A Cdk9-PP1 switch regulates the elongation-termination transition of RNA polymerase II |
| Organism |
Schizosaccharomyces pombe |
| Experiment type |
Other
Genome binding/occupancy profiling by high throughput sequencing
|
| Summary |
The end of the RNA polymerase II (Pol II) transcription cycle is strictly regulated to ensure proper mRNA maturation and prevent interference between neighboring genes. Pol II slowing downstream of the cleavage and polyadenylation signal (CPS) leads to recruitment of cleavage and polyadenylation factors and termination, but how this chain of events is initiated remains unclear. In a chemical-genetic screen we identified protein phosphatase 1 (PP1) isoforms as substrates of human positive transcription elongation factor b (P-TEFb), the cyclin-dependent kinase 9 (Cdk9)-cyclin T1 complex. Here we show that Cdk9 and PP1 govern phosphorylation of the conserved transcription factor Spt5 in the fission yeast Schizosaccharomyces pombe. Cdk9 phosphorylates both Spt5 and a negative regulatory site on the PP1 isoform Dis2. Sites phosphorylated by Cdk9 in the Spt5 carboxy-terminal domain (CTD) are dephosphorylated by Dis2 in vitro, and Cdk9 inhibition in vivo leads to rapid Spt5 dephosphorylation that is retarded by concurrent Dis2 inactivation. Chromatin immunoprecipitation and sequencing (ChIP-seq) analysis indicates that Spt5 is dephosphorylated as transcription complexes traverse the CPS, prior to or concomitant with Pol II pausing. A Dis2-inactivating mutation stabilizes Spt5 phosphorylation (pSpt5) on chromatin, promotes transcription beyond the normal termination zone detected by precision run-on transcription and sequencing (PRO-seq), and is suppressed by ablation of Cdk9 target sites in Spt5. These results support a model whereby the transition of Pol II from elongation to termination is regulated by opposing activities of Cdk9 and Dis2 towards their common substrate Spt5—a bistable switch analogous to a Cdk1-PP1 module that controls exit from mitosis.
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| Overall design |
PRO-seq: Data represent 20 different treatments of various S. pombe strains performed in biological replicates. There are processed files for both replicates and combined replicate data for each experiment.
ChIP-seq: Data represent 6 different ChIP-seq experiments in S. pombe, performed in biological replicates. There are raw and processed files for all biological replicates.
ChIP-seq: Data represent 16 different ChIP-seq experiments in S. pombe, performed in biological replicates. There are raw and processed files for all biological replicates.
PRO-seq: Experiment was done in five different S. pombe strains as biological duplicates. There are processed files for both replicates and combined replicate data for each sample.
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| |
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| Contributor(s) |
Parua PK, Booth GT, Sanso M, Tanny J |
| Citation(s) |
29899453, 32496538 |
| |
| Submission date |
Aug 11, 2017 |
| Last update date |
Jul 25, 2021 |
| Contact name |
PABITRA K PARUA |
| E-mail(s) |
pabitra.parua@einsteinmed.eduy
|
| Phone |
7184304284
|
| Organization name |
Albert Einstein College of Medicine
|
| Department |
Molecular Pharmacology
|
| Lab |
Forch-236
|
| Street address |
1300 Morris Park Avenue, Forchheimer 236, Molecular Pharmacology
|
| City |
Bronx |
| State/province |
NY |
| ZIP/Postal code |
10461 |
| Country |
USA |
| |
|
| Platforms (2) |
| GPL13988 |
Illumina HiSeq 2000 (Schizosaccharomyces pombe) |
| GPL20584 |
Illumina NextSeq 500 (Schizosaccharomyces pombe) |
|
| Samples (52)
|
|
| Relations |
| BioProject |
PRJNA398083 |
| SRA |
SRP115327 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE102590_7772_7157_43149_HVLYCBGXY_pombe_18C_Cdk9as_DMSO_COMBINED_GGCTAC_R1_pombe_SpikeNormed_minus.bw |
9.3 Mb |
(ftp)(http) |
BW |
| GSE102590_7772_7157_43149_HVLYCBGXY_pombe_18C_Cdk9as_DMSO_COMBINED_GGCTAC_R1_pombe_SpikeNormed_plus.bw |
9.1 Mb |
(ftp)(http) |
BW |
| GSE102590_7772_7157_43150_HVLYCBGXY_pombe_18C_Cdk9as_3MBPP1_COMBINED_CTTGTA_R1_pombe_SpikeNormed_minus.bw |
8.2 Mb |
(ftp)(http) |
BW |
| GSE102590_7772_7157_43150_HVLYCBGXY_pombe_18C_Cdk9as_3MBPP1_COMBINED_CTTGTA_R1_pombe_SpikeNormed_plus.bw |
8.2 Mb |
(ftp)(http) |
BW |
| GSE102590_7772_7157_43151_HVLYCBGXY_pombe_18C_dis2ts_DMSO_COMBINED_ATTCCT_R1_pombe_SpikeNormed_minus.bw |
5.7 Mb |
(ftp)(http) |
BW |
| GSE102590_7772_7157_43151_HVLYCBGXY_pombe_18C_dis2ts_DMSO_COMBINED_ATTCCT_R1_pombe_SpikeNormed_plus.bw |
5.6 Mb |
(ftp)(http) |
BW |
| GSE102590_7772_7157_43152_HVLYCBGXY_pombe_18C_dis2ts_3MBPP1_COMBINED_CAAAAG_R1_pombe_SpikeNormed_minus.bw |
5.4 Mb |
(ftp)(http) |
BW |
| GSE102590_7772_7157_43152_HVLYCBGXY_pombe_18C_dis2ts_3MBPP1_COMBINED_CAAAAG_R1_pombe_SpikeNormed_plus.bw |
5.3 Mb |
(ftp)(http) |
BW |
| GSE102590_7772_7157_43153_HVLYCBGXY_pombe_30C_dis2ts_DMSO_COMBINED_CAACTA_R1_pombe_SpikeNormed_minus.bw |
6.6 Mb |
(ftp)(http) |
BW |
| GSE102590_7772_7157_43153_HVLYCBGXY_pombe_30C_dis2ts_DMSO_COMBINED_CAACTA_R1_pombe_SpikeNormed_plus.bw |
6.5 Mb |
(ftp)(http) |
BW |
| GSE102590_7772_7157_43154_HVLYCBGXY_pombe_30C_dis2ts_3MBPP1_COMBINED_CACCGG_R1_pombe_SpikeNormed_minus.bw |
5.5 Mb |
(ftp)(http) |
BW |
| GSE102590_7772_7157_43154_HVLYCBGXY_pombe_30C_dis2ts_3MBPP1_COMBINED_CACCGG_R1_pombe_SpikeNormed_plus.bw |
5.4 Mb |
(ftp)(http) |
BW |
| GSE102590_7772_7157_43155_HVLYCBGXY_pombe_18C_Cdk9as_dis2ts_DMSO_COMBINED_CACGAT_R1_pombe_SpikeNormed_minus.bw |
5.7 Mb |
(ftp)(http) |
BW |
| GSE102590_7772_7157_43155_HVLYCBGXY_pombe_18C_Cdk9as_dis2ts_DMSO_COMBINED_CACGAT_R1_pombe_SpikeNormed_plus.bw |
5.6 Mb |
(ftp)(http) |
BW |
| GSE102590_7772_7157_43156_HVLYCBGXY_pombe_18C_Cdk9as_dis2ts_3MBPP1_COMBINED_CACTCA_R1_pombe_SpikeNormed_minus.bw |
6.5 Mb |
(ftp)(http) |
BW |
| GSE102590_7772_7157_43156_HVLYCBGXY_pombe_18C_Cdk9as_dis2ts_3MBPP1_COMBINED_CACTCA_R1_pombe_SpikeNormed_plus.bw |
6.5 Mb |
(ftp)(http) |
BW |
| GSE102590_7772_7157_43157_HVLYCBGXY_pombe_30C_Cdk9as_dis2ts_DMSO_COMBINED_CAGGCG_R1_pombe_SpikeNormed_minus.bw |
5.5 Mb |
(ftp)(http) |
BW |
| GSE102590_7772_7157_43157_HVLYCBGXY_pombe_30C_Cdk9as_dis2ts_DMSO_COMBINED_CAGGCG_R1_pombe_SpikeNormed_plus.bw |
5.4 Mb |
(ftp)(http) |
BW |
| GSE102590_7772_7157_43158_HVLYCBGXY_pombe_30C_Cdk9as_dis2ts_3MBPP1_COMBINED_CATGGC_R1_pombe_SpikeNormed_minus.bw |
5.4 Mb |
(ftp)(http) |
BW |
| GSE102590_7772_7157_43158_HVLYCBGXY_pombe_30C_Cdk9as_dis2ts_3MBPP1_COMBINED_CATGGC_R1_pombe_SpikeNormed_plus.bw |
5.3 Mb |
(ftp)(http) |
BW |
| GSE102590_ChIPSeq_Spt5_Myc_Combined.bedgraph.gz |
93.3 Mb |
(ftp)(http) |
BEDGRAPH |
| GSE102590_ChIPSeq_pSpt5_Combined.bedgraph.gz |
109.8 Mb |
(ftp)(http) |
BEDGRAPH |
| GSE102590_Concatenate_pp_chipseq_MACS2_dis2cs_IgG.bigwig |
42.0 Mb |
(ftp)(http) |
BIGWIG |
| GSE102590_Concatenate_pp_chipseq_MACS2_dis2cs_myc.bigwig |
78.4 Mb |
(ftp)(http) |
BIGWIG |
| GSE102590_Concatenate_pp_chipseq_MACS2_dis2cs_pSpt5.bigwig |
85.9 Mb |
(ftp)(http) |
BIGWIG |
| GSE102590_Concatenate_pp_chipseq_MACS2_wt_IgG.bigwig |
42.6 Mb |
(ftp)(http) |
BIGWIG |
| GSE102590_Concatenate_pp_chipseq_MACS2_wt_myc.bigwig |
80.4 Mb |
(ftp)(http) |
BIGWIG |
| GSE102590_Concatenate_pp_chipseq_MACS2_wt_pSpt5.bigwig |
84.7 Mb |
(ftp)(http) |
BIGWIG |
| GSE102590_RAW.tar |
1.1 Gb |
(http)(custom) |
TAR (of BEDGRAPH, BIGWIG, BW) |
| GSE102590_ePROseq_Pombe_Fisher_WT_COMBINED_pombe_spikeNormed_minus.bw |
2.8 Mb |
(ftp)(http) |
BW |
| GSE102590_ePROseq_Pombe_Fisher_WT_COMBINED_pombe_spikeNormed_plus.bw |
2.8 Mb |
(ftp)(http) |
BW |
| GSE102590_ePROseq_Pombe_Fisher_dis2Del_COMBINED_pombe_spikeNormed_minus.bw |
3.0 Mb |
(ftp)(http) |
BW |
| GSE102590_ePROseq_Pombe_Fisher_dis2Del_COMBINED_pombe_spikeNormed_plus.bw |
2.9 Mb |
(ftp)(http) |
BW |
| GSE102590_ePROseq_Pombe_Fisher_dis2_11_COMBINED_pombe_spikeNormed_minus.bw |
4.7 Mb |
(ftp)(http) |
BW |
| GSE102590_ePROseq_Pombe_Fisher_dis2_11_COMBINED_pombe_spikeNormed_plus.bw |
4.7 Mb |
(ftp)(http) |
BW |
| GSE102590_ePROseq_Pombe_Fisher_dis2_T316A_COMBINED_pombe_spikeNormed_minus.bw |
4.2 Mb |
(ftp)(http) |
BW |
| GSE102590_ePROseq_Pombe_Fisher_dis2_T316A_COMBINED_pombe_spikeNormed_plus.bw |
4.1 Mb |
(ftp)(http) |
BW |
| GSE102590_ePROseq_Pombe_Fisher_dis2_T316D_COMBINED_pombe_spikeNormed_minus.bw |
4.6 Mb |
(ftp)(http) |
BW |
| GSE102590_ePROseq_Pombe_Fisher_dis2_T316D_COMBINED_pombe_spikeNormed_plus.bw |
4.5 Mb |
(ftp)(http) |
BW |
| GSE102590_pp_chipseq_IgGSubtracted_dis2cs_myc.bigwig |
7.8 Mb |
(ftp)(http) |
BIGWIG |
| GSE102590_pp_chipseq_IgGSubtracted_dis2cs_pSpt5.bigwig |
7.9 Mb |
(ftp)(http) |
BIGWIG |
| GSE102590_pp_chipseq_IgGSubtracted_wt_myc.bigwig |
7.9 Mb |
(ftp)(http) |
BIGWIG |
| GSE102590_pp_chipseq_IgGSubtracted_wt_pSpt5.bigwig |
7.9 Mb |
(ftp)(http) |
BIGWIG |
SRA Run Selector |
| Processed data are available on Series record |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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