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Pol II and H3K27Ac occupancy of the human genome in MRC5 cells

ABSTRACT: Pol II and H3K27Ac occupancy of the human genome in MRC5 cells

PROVIDER: PRJNA719831 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR14145945_1.fastq.gz	Fastqsanger.gz
	SRR14145945_2.fastq.gz	Fastqsanger.gz
	SRR14145946_1.fastq.gz	Fastqsanger.gz
	SRR14145946_2.fastq.gz	Fastqsanger.gz
	SRR14145947_1.fastq.gz	Fastqsanger.gz

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Pol II and H3K27Ac occupancy of the human genome in MRC5 cells

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Project description:We used RNA Pol II-CTD occupancy assay to identify mRNA isoform switch in mutant animals compared to wild type animals. Examination of RNA Pol II-CTD Ser5P recognized by 4H8 in wild type, sydn-1(0), ssup-72(0) and ssup-72(0);sydn-1(0)

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Project description:ATP-dependent chromatin remodelers regulate chromatin structure during multiple stages of transcription. We report that RSC, an essential chromatin remodeler, is recruited to the open reading frames (ORFs) of actively transcribed genes genome-wide, suggesting a role for RSC in regulating transcription elongation. Consistent with such a role, Pol II occupancy in the ORFs of weakly transcribed genes is drastically reduced upon depletion of the RSC catalytic subunit Sth1. RSC inactivation also reduced histone H3 occupancy across transcribed regions. Remarkably, the strongest effects on Pol II and H3 occupancy were confined to the genes displaying the greatest RSC ORF enrichment. Additionally, RSC recruitment to the ORF requires the activities of the SAGA and NuA4 HAT complexes and is aided by the activities of the Pol II CTD Ser2 kinases Bur1 and Ctk1. Overall, our findings strongly implicate ORF-associated RSC in governing Pol II function and in maintaining chromatin structure over transcribed regions. ChIP-chip experiments to measure Sth1, Rpb3 and H3 occupancy in WT and various mutants (histone acetyltransferase and Pol II CTD kinase mutants). The histone H3 and Rpb3 occupancy were also measured in cells upon Sth1 depletion.

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