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

Molecular and Cellular Biology
Yong ShenJörg Bungert

Abstract

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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Citations

Mar 21, 2021·Current Opinion in Hematology·Marlies P Rossmann, Leonard I Zon

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Datasets Mentioned

BETA
GSM2695565
GSM1816074

Methods Mentioned

BETA
acetylation
immunoprecipitation
electrophoretic mobility shift
ChIP
PCR
ChIP-seq
Hi-C
transfection
electrophoresis
X-ray

Software Mentioned

Galaxy
RepeatMasker
BigWig
GRO
Integrative Genomics Viewer ( IGV
GraphPad Prism
Bowtie 2
Integrative Genomics Viewer ( IGV )

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