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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...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Microbiology
2018
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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. |
format | Online Article Text |
id | pubmed-6146835 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | American Society for Microbiology |
record_format | MEDLINE/PubMed |
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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