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Identification of a Novel Enhancer/Chromatin Opening Element Associated with High-Level γ-Globin Gene Expression

The organization of the five β-type globin genes on chromosome 11 reflects the timing of expression during erythroid cell development, with the embryonic ε-globin gene being located at the 5′ end, followed by the two fetal γ-globin genes, and with the adult β- and δ-globin genes being located at the...

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Autores principales: Shen, Yong, Bassett, MacLean A., Gurumurthy, Aishwarya, Nar, Rukiye, Knudson, Isaac J., Guy, Cameron R., Perez, Alex, Mellen, Russell W., Ikeda, Masatoshi, Hossain, Mir A., Huang, Suming, Igarashi, Kazuhiko, Bungert, Jörg
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Microbiology 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6146835/
https://www.ncbi.nlm.nih.gov/pubmed/30012865
http://dx.doi.org/10.1128/MCB.00197-18
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author Shen, Yong
Bassett, MacLean A.
Gurumurthy, Aishwarya
Nar, Rukiye
Knudson, Isaac J.
Guy, Cameron R.
Perez, Alex
Mellen, Russell W.
Ikeda, Masatoshi
Hossain, Mir A.
Huang, Suming
Igarashi, Kazuhiko
Bungert, Jörg
author_facet Shen, Yong
Bassett, MacLean A.
Gurumurthy, Aishwarya
Nar, Rukiye
Knudson, Isaac J.
Guy, Cameron R.
Perez, Alex
Mellen, Russell W.
Ikeda, Masatoshi
Hossain, Mir A.
Huang, Suming
Igarashi, Kazuhiko
Bungert, Jörg
author_sort Shen, Yong
collection PubMed
description The organization of the five β-type globin genes on chromosome 11 reflects the timing of expression during erythroid cell development, with the embryonic ε-globin gene being located at the 5′ end, followed by the two fetal γ-globin genes, and with the adult β- and δ-globin genes being located at the 3′ end. Here, we functionally characterized a DNase I-hypersensitive site (HS) located 4 kb upstream of the Gγ-globin gene (HBG-4kb HS). This site is occupied by transcription factors USF1, USF2, EGR1, MafK, and NF-E2 in the human erythroleukemia cell line K562 and exhibits histone modifications typical for enhancers. We generated a synthetic zinc finger (ZF) DNA-binding domain targeting the HBG-4kb HS (HBG-4kb ZF). The HBG-4kb ZF interacted with the target site in vitro and in the context of cells with a high affinity and specificity. Direct delivery of the HBG-4kb ZF to K562 and primary human erythroid cells caused a reduction in γ-globin gene expression which was associated with decreased binding of transcription factors and active histone marks at and downstream of the HS. The data demonstrate that the HBG-4kb HS is important for fetal globin production and suggest that it may act by opening chromatin in a directional manner.
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spelling pubmed-61468352018-09-28 Identification of a Novel Enhancer/Chromatin Opening Element Associated with High-Level γ-Globin Gene Expression Shen, Yong Bassett, MacLean A. Gurumurthy, Aishwarya Nar, Rukiye Knudson, Isaac J. Guy, Cameron R. Perez, Alex Mellen, Russell W. Ikeda, Masatoshi Hossain, Mir A. Huang, Suming Igarashi, Kazuhiko Bungert, Jörg Mol Cell Biol Research Article The organization of the five β-type globin genes on chromosome 11 reflects the timing of expression during erythroid cell development, with the embryonic ε-globin gene being located at the 5′ end, followed by the two fetal γ-globin genes, and with the adult β- and δ-globin genes being located at the 3′ end. Here, we functionally characterized a DNase I-hypersensitive site (HS) located 4 kb upstream of the Gγ-globin gene (HBG-4kb HS). This site is occupied by transcription factors USF1, USF2, EGR1, MafK, and NF-E2 in the human erythroleukemia cell line K562 and exhibits histone modifications typical for enhancers. We generated a synthetic zinc finger (ZF) DNA-binding domain targeting the HBG-4kb HS (HBG-4kb ZF). The HBG-4kb ZF interacted with the target site in vitro and in the context of cells with a high affinity and specificity. Direct delivery of the HBG-4kb ZF to K562 and primary human erythroid cells caused a reduction in γ-globin gene expression which was associated with decreased binding of transcription factors and active histone marks at and downstream of the HS. The data demonstrate that the HBG-4kb HS is important for fetal globin production and suggest that it may act by opening chromatin in a directional manner. American Society for Microbiology 2018-09-14 /pmc/articles/PMC6146835/ /pubmed/30012865 http://dx.doi.org/10.1128/MCB.00197-18 Text en Copyright © 2018 Shen et al. https://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research Article
Shen, Yong
Bassett, MacLean A.
Gurumurthy, Aishwarya
Nar, Rukiye
Knudson, Isaac J.
Guy, Cameron R.
Perez, Alex
Mellen, Russell W.
Ikeda, Masatoshi
Hossain, Mir A.
Huang, Suming
Igarashi, Kazuhiko
Bungert, Jörg
Identification of a Novel Enhancer/Chromatin Opening Element Associated with High-Level γ-Globin Gene Expression
title Identification of a Novel Enhancer/Chromatin Opening Element Associated with High-Level γ-Globin Gene Expression
title_full Identification of a Novel Enhancer/Chromatin Opening Element Associated with High-Level γ-Globin Gene Expression
title_fullStr Identification of a Novel Enhancer/Chromatin Opening Element Associated with High-Level γ-Globin Gene Expression
title_full_unstemmed Identification of a Novel Enhancer/Chromatin Opening Element Associated with High-Level γ-Globin Gene Expression
title_short Identification of a Novel Enhancer/Chromatin Opening Element Associated with High-Level γ-Globin Gene Expression
title_sort identification of a novel enhancer/chromatin opening element associated with high-level γ-globin gene expression
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6146835/
https://www.ncbi.nlm.nih.gov/pubmed/30012865
http://dx.doi.org/10.1128/MCB.00197-18
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