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The environmentally-regulated interplay between three-dimensional chromatin organisation and transcription of the Escherichia coli proVWX operon

ABSTRACT: The environmentally-regulated interplay between three-dimensional chromatin organisation and transcription of the Escherichia coli proVWX operon

PROVIDER: PRJNA885782 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR21765562.fastq.gz	Fastqsanger.gz
	SRR21765562_1.fastq.gz	Fastqsanger.gz
	SRR21765562_2.fastq.gz	Fastqsanger.gz
	SRR21765563.fastq.gz	Fastqsanger.gz
	SRR21765563_1.fastq.gz	Fastqsanger.gz

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The environmentally-regulated interplay between three-dimensional chromatin organisation and transcription of the Escherichia coli proVWX operon

Project description:Nucleoid associated proteins maintain the architecture of bacterial chromosomes and regulate gene expression. Thus, their role as transcription factors may involve three-dimensional chromosome re-modelling. Here, we provide the first in vivo evidence supporting this hypothesis. Using RT-qPCR and 3C-qPCR, we show that activation of the H-NS-regulated, osmosensitive proVWX operon of Escherichia coli in response to a hyperosmotic shock involves the destabilization of H-NS-mediated bridges anchored between the proVWX downstream and upstream regulatory elements (DRE and URE), and between the DRE and ygaY that lies immediately downstream of proVWX. The re-establishment of these bridges in cells adapted to hyperosmolarity represses the operon. While the DRE—H-NS—URE bridge is disrupted by hyperosmolarity, the DRE—H-NS—ygaY bridge weakens and requires active transcription to be dismantled. H-NS and H-NS-like proteins are wide-spread amongst bacteria, suggesting that structural regulation of expression may be a typical feature of transcriptional control in bacteria.

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The environmentally-regulated interplay between local three-dimensional chromatin organisation and transcription of proVWX in E. coli.

Project description:Nucleoid associated proteins (NAPs) maintain the architecture of bacterial chromosomes and regulate gene expression. Thus, their role as transcription factors may involve three-dimensional chromosome re-organisation. While this model is supported by in vitro studies, direct in vivo evidence is lacking. Here, we use RT-qPCR and 3C-qPCR to study the transcriptional and architectural profiles of the H-NS (histone-like nucleoid structuring protein)-regulated, osmoresponsive proVWX operon of Escherichia coli at different osmolarities and provide in vivo evidence for transcription regulation by NAP-mediated chromosome re-modelling in bacteria. By consolidating our in vivo investigations with earlier in vitro and in silico studies that provide mechanistic details of how H-NS re-models DNA in response to osmolarity, we report that activation of proVWX in response to a hyperosmotic shock involves the destabilization of H-NS-mediated bridges anchored between the proVWX downstream and upstream regulatory elements (DRE and URE), and between the DRE and ygaY that lies immediately downstream of proVWX. The re-establishment of these bridges upon adaptation to hyperosmolarity represses the operon. Our results also reveal additional structural features associated with changes in proVWX transcript levels such as the decompaction of local chromatin upstream of the operon, highlighting that further complexity underlies the regulation of this model operon. H-NS and H-NS-like proteins are wide-spread amongst bacteria, suggesting that chromosome re-modelling may be a typical feature of transcriptional control in bacteria.

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Self-organisation of human pluripotent stem cells into three-dimensional model of post-implantation development

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Spatial organisation of the X inactivation center

Project description:We report the application of Chromosome Conformation Capture Carbon-copy (5C) to a 4.5 Mb stretch of the mouse X chromosome encompassing the X inactivation center locus. We uncover a series of discrete 200kb-1Mb topologically associating domains (TADs). These align with several domain-wide epigenomic features as well as co-regulated gene clusters. 5C analysis in EED and G9A mutants reveal that this segmental organisation in TADs does not relie on the underlying H3K27me3 or H3K9me2 blocks. Deletion of a boundary between two TADs leads to ectopic chromosomal contacts between them. Analysis of mESCs, mNPCs and MEFs suggest that the positioning of TADs on the chromosome is stable during cell differentiation though their internal organisation changes. Comparison of male (XY) and female (XX) differentiated cells highlights that the long-range chromosomal contacts within TADs are dampened on the inactive X compared to the active X. 5C oligonucleotides were designed around HindIII restriction site following an alternative scheme

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A histone hyperacetylation pulse induces cellular memory of gene expression and 3-dimensional genome folding [microC_3]

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A histone hyperacetylation pulse induces cellular memory of gene expression and 3-dimensional genome folding [microC_2]

Project description:Epigenetic memory enables the stable propagation of gene expression patterns in response to transient developmental and environmental stimuli. Although 3-dimensional (3D) organisation is emerging as a key regulator of genome function due its role in gene expression control, it is unknown whether it contributes to cellular memory. Here, we establish that acute perturbation of the epigenome can induce cellular memory of gene expression in mouse embryonic stem cells (mESCs). Specifically, we uncover how a pulse of histone deacetylase inhibition translates to changes in the histone acetylation and methylation landscape, as well as global and local genome folding. While most epigenomic and transcriptional changes are readily reversible once the perturbation is removed, architectural features are not fully reestablished. Upon a second transient pulse of hyperacetylation, hundreds of genes maintain their dysregulated state. Using ultra-deep Micro-C, we associate memory of gene expression with enhancer-promoter contacts and repressive chromatin topology mediated by Polycomb. These results uncover the complex interplay between different epigenetic regulatory layers and establish a novel link between genome folding and cellular memory.

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