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RNA Polymerase II pausing temporally coordinates cell cycle progression and erythroid differentiation [CRISPR]

ABSTRACT: RNA Polymerase II pausing temporally coordinates cell cycle progression and erythroid differentiation [CRISPR]

PROVIDER: PRJNA885860 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR21771600_1.fastq.gz	Fastqsanger.gz
	SRR21771600_2.fastq.gz	Fastqsanger.gz
	SRR21771601_1.fastq.gz	Fastqsanger.gz
	SRR21771601_2.fastq.gz	Fastqsanger.gz
	SRR21771602_1.fastq.gz	Fastqsanger.gz

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RNA Polymerase II pausing temporally coordinates cell cycle progression and erythroid differentiation

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RNA Polymerase II pausing temporally coordinates cell cycle progression and erythroid differentiation [CRISPR]

Project description:The controlled release of promoter-proximal RNA polymerase II (Pol II) pausing into productive elongation is a major step in gene regulation. Yet, functionally analyzing Pol II pausing is difficult because the factors that regulate pause release are generally essential. Here, we identified heterozygous loss-of-function mutations in SUPT5H, encoding SPT5, in individuals with β-thalassemia trait unlinked to the β-globin locus. Recapitulating the pathogenic mutations in human erythroid cells or acute treatment with transcription inhibitors replicates the patient phenotype of altered globin gene expression. Furthermore, in SUPT5H-edited cells, Pol II pause release was globally disrupted. We observed dynamic patterns of pausing at cell cycle and erythroid differentiation genes at the transition from progenitors to precursors during erythropoiesis. At this same transition, in SUPT5H-edited cells, we observed delayed differentiation and altered cell cycle kinetics, as well as a delay in the onset of gene expression programs. Therefore, hindering pause release perturbs cell cycle and delays differentiation at key stages of erythropoiesis, revealing a role for Pol II pausing in the temporal coordination between the cell cycle and differentiation.

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RNA Polymerase II pausing temporally coordinates cell cycle progression and erythroid differentiation

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Polymerase II pausing licenses cell-cycle progression

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Polymerase II pausing licenses cell-cycle progression (ChIP-Seq)

Project description:RNA polymerase II promoter-proximal pausing is orchestrated by a ribonucleoprotein complex scaffolded by the noncoding RNA Rn7sk. However, how this interruption of transcription is mechanistically linked to RNA production remains largely unknown. Here, we show that forcing the pause release by germ-line deletion of Rn7sk was embryonic lethal, yet conditional deletion of Rn7sk enhanced stem cell differentiation in skin. To explore the immediate transcriptional mechanisms underpinning enhanced differentiation, we metabolically labelled newly-synthesized RNAs after Rn7sk deletion. Unexpectedly, forced pause release robustly repressed transcription specifically at cell cycle regulators, in the absence of chromatin remodeling at promoters and enhancers. Our results indicate that polymerase pausing affords the core elongation machinery time to properly assemble, and forced elongation triggers splicing defects and nuclear RNA decay. Cell cycle regulators appear highly sensitive to mis-regulation of the elongation machinery due to unique genomic features of high promoter accessibility and low GC–content in the gene body. Transcriptional pausing thus serves as a rate-limiting step in controlling cell division.

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