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Tandem CTCF sites function as insulators to balance spatial chromatin contacts and topological enhancer-promoter selection

Identifieur interne : 000140 ( new/Analysis ); précédent : 000139; suivant : 000141

Tandem CTCF sites function as insulators to balance spatial chromatin contacts and topological enhancer-promoter selection

Auteurs : Zhilian Jia [République populaire de Chine] ; Jingwei Li [République populaire de Chine] ; Xiao Ge [République populaire de Chine] ; Yonghu Wu [République populaire de Chine] ; Ya Guo [République populaire de Chine] ; Qiang Wu [République populaire de Chine]

Source :

RBID : PMC:7087399

Abstract

Background

CTCF is a key insulator-binding protein, and mammalian genomes contain numerous CTCF sites, many of which are organized in tandem.

Results

Using CRISPR DNA-fragment editing, in conjunction with chromosome conformation capture, we find that CTCF sites, if located between enhancers and promoters in the protocadherin (Pcdh) and β-globin clusters, function as an enhancer-blocking insulator by forming distinct directional chromatin loops, regardless whether enhancers contain CTCF sites or not. Moreover, computational simulation in silico and genetic deletions in vivo as well as dCas9 blocking in vitro revealed balanced promoter usage in cell populations and stochastic monoallelic expression in single cells by large arrays of tandem CTCF sites in the Pcdh and immunoglobulin heavy chain (Igh) clusters. Furthermore, CTCF insulators promote, counter-intuitively, long-range chromatin interactions with distal directional CTCF sites, consistent with the cohesin “loop extrusion” model. Finally, gene expression levels are negatively correlated with CTCF insulators located between enhancers and promoters on a genome-wide scale. Thus, single CTCF insulators ensure proper enhancer insulation and promoter activation while tandem CTCF topological insulators determine balanced spatial contacts and promoter choice.

Conclusions

These findings have interesting implications on the role of topological chromatin insulators in 3D genome folding and developmental gene regulation.


Url:
DOI: 10.1186/s13059-020-01984-7
PubMed: NONE
PubMed Central: 7087399


Affiliations:


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PMC:7087399

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<title>Background</title>
<p id="Par1">CTCF is a key insulator-binding protein, and mammalian genomes contain numerous CTCF sites, many of which are organized in tandem.</p>
</sec>
<sec>
<title>Results</title>
<p id="Par2">Using CRISPR DNA-fragment editing, in conjunction with chromosome conformation capture, we find that CTCF sites, if located between enhancers and promoters in the protocadherin (
<italic>Pcdh</italic>
) and
<italic>β-globin</italic>
clusters, function as an enhancer-blocking insulator by forming distinct directional chromatin loops, regardless whether enhancers contain CTCF sites or not. Moreover, computational simulation in silico and genetic deletions in vivo as well as dCas9 blocking in vitro revealed balanced promoter usage in cell populations and stochastic monoallelic expression in single cells by large arrays of tandem CTCF sites in the
<italic>Pcdh</italic>
and immunoglobulin heavy chain (
<italic>Igh</italic>
) clusters. Furthermore, CTCF insulators promote, counter-intuitively, long-range chromatin interactions with distal directional CTCF sites, consistent with the cohesin “loop extrusion” model. Finally, gene expression levels are negatively correlated with CTCF insulators located between enhancers and promoters on a genome-wide scale. Thus, single CTCF insulators ensure proper enhancer insulation and promoter activation while tandem CTCF topological insulators determine balanced spatial contacts and promoter choice.</p>
</sec>
<sec>
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<p id="Par3">These findings have interesting implications on the role of topological chromatin insulators in 3D genome folding and developmental gene regulation.</p>
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