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

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

PROVIDER: PRJNA885863 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR21771700_1.fastq.gz	Fastqsanger.gz
	SRR21771700_2.fastq.gz	Fastqsanger.gz
	SRR21771701_1.fastq.gz	Fastqsanger.gz
	SRR21771701_2.fastq.gz	Fastqsanger.gz
	SRR21771702_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 contributes to maintain chromatin organization in erythrocytes [RNA-seq]

Project description:Chicken erythrocytes are nucleated cells often referred to as transcriptionally inactive, although the epigenetic changes and chromatin remodeling that mediate transcriptional repression and the extent of gene silencing during avian terminal erythroid differentiation are not fully understood. Here we characterized the changes in gene expression, chromatin accessibility, genome organization, and chromatin nuclear disposition during the terminal stages of erythropoiesis in chicken and found a complex chromatin reorganization at different genomic scales. We identified a robust decrease in transcription in erythrocytes. Nevertheless, a set of genes maintains their expression in erythrocytes, including genes involved in RNA pol II promoter-proximal pausing. Erythrocytes exhibit an inverted nuclear architecture and reposition euchromatin towards the nuclear periphery together with the paused RNA polymerase. In erythrocytes, chromatin domains are partially lost genome-wide except at mini domains retained around paused promoters. Our results suggest that promoter-proximal pausing of the RNA pol II participates in the transcriptional regulation of the erythroid genome and highlight the role of RNA polymerase in the maintenance of local chromatin organization.

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