Identification of a novel enhancer/chromatin opening element associated with high-level -globin gene expression

Yong Shen, MacLean A. Bassett, Aishwarya Gurumurthy, Rukiye Nar, Isaac J. Knudson, Cameron R. Guy, Alex Perez, Russell W. Mellen, Masatoshi Ikeda, Mir A. Hossain, Suming Huang, Kazuhiko Igarashi, Jörg Bungert

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

The organization of the five -type globin genes on chromosome 11 reflects the timing of expression during erythroid cell development, with the embryonic -globin gene being located at the 5= end, followed by the two fetal -globin genes, and with the adult - and -globin genes being located at the 3= end. Here, we functionally characterized a DNase I-hypersensitive site (HS) located 4 kb upstream of the G-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 -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.

Original languageEnglish (US)
Article numbere00197
JournalMolecular and cellular biology
Volume38
Issue number19
DOIs
StatePublished - Oct 1 2018

Fingerprint

Globins
Chromatin
Gene Expression
Zinc Fingers
Histone Code
Erythroid Cells
Genes
Transcription Factors
Leukemia, Erythroblastic, Acute
Chromosomes, Human, Pair 11
Deoxyribonuclease I
Embryonic Development
Cell Line
DNA

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

Cite this

Shen, Y., Bassett, M. A., Gurumurthy, A., Nar, R., Knudson, I. J., Guy, C. R., ... Bungert, J. (2018). Identification of a novel enhancer/chromatin opening element associated with high-level -globin gene expression. Molecular and cellular biology, 38(19), [e00197]. https://doi.org/10.1128/MCB.00197-18
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 -globin gene expression. In: Molecular and cellular biology. 2018 ; Vol. 38, No. 19.
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Shen, Y, Bassett, MA, Gurumurthy, A, Nar, R, Knudson, IJ, Guy, CR, Perez, A, Mellen, RW, Ikeda, M, Hossain, MA, Huang, S, Igarashi, K & Bungert, J 2018, 'Identification of a novel enhancer/chromatin opening element associated with high-level -globin gene expression', Molecular and cellular biology, vol. 38, no. 19, e00197. https://doi.org/10.1128/MCB.00197-18

Identification of a novel enhancer/chromatin opening element associated with high-level -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.

In: Molecular and cellular biology, Vol. 38, No. 19, e00197, 01.10.2018.

Research output: Contribution to journalArticle

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T1 - Identification of a novel enhancer/chromatin opening element associated with high-level -globin gene expression

AU - Shen, Yong

AU - Bassett, MacLean A.

AU - Gurumurthy, Aishwarya

AU - Nar, Rukiye

AU - Knudson, Isaac J.

AU - Guy, Cameron R.

AU - Perez, Alex

AU - Mellen, Russell W.

AU - Ikeda, Masatoshi

AU - Hossain, Mir A.

AU - Huang, Suming

AU - Igarashi, Kazuhiko

AU - Bungert, Jörg

PY - 2018/10/1

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N2 - The organization of the five -type globin genes on chromosome 11 reflects the timing of expression during erythroid cell development, with the embryonic -globin gene being located at the 5= end, followed by the two fetal -globin genes, and with the adult - and -globin genes being located at the 3= end. Here, we functionally characterized a DNase I-hypersensitive site (HS) located 4 kb upstream of the G-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 -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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