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Identification of a novel enhancer/chromatin opening element associated with high‐level γ‐globin gene expression
Author(s) -
Bassett Maclean,
Shen Yong,
Hossain Mir,
Guy Cameron,
Nar Rukiye,
Gurumurthy Aishwarya,
Bungert Jörg
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.lb9
Subject(s) - microbiology and biotechnology , chromatin , enhancer , biology , gene , globin , histone , locus control region , gene expression , transcription (linguistics) , genetics , linguistics , philosophy
The human β‐globin gene locus is located on chromosome 11 and constitutes five β‐type globin genes that are organized in a manner reflecting their expression during development; an embryonic ɛ‐globin gene located at the 5′ end, followed by the two fetal γ‐globin genes, and the adult β‐ and δ‐globin genes at the 3′ end. A novel DNase I hypersensitive site (HS) located 4 kb upstream of the Gγ‐globin gene (HBG‐4kb HS) was targeted as a candidate for affecting expression of γ‐globin. In the human erythroleukemia cell line K562, this site is occupied by transcription factors USF1, USF2, EGR1, MafK, and NF‐E2; and exhibits histone modifications typical of enhancer regions. A synthetic zinc finger DNA‐binding domain (ZF‐DBD) was generated to target the HBG‐4kb HS. The HBG‐4kb ZF‐DBD interacted with the target site in vitro and in the context of cells with high affinity and specificity. Direct delivery of HBG‐4kb ZF‐DBD to K562 and primary human erythroid cells resulted in a reduced association of the transcription factors identified and active histone marks present at and downstream of the HS site. Reduction in γ‐globin gene expression was observed following introduction of the HBG‐4kb ZF‐DBD. The data demonstrate that the HBG‐4kb HS site affects fetal γ‐globin gene expression and extrapolation suggests that the site may act by opening chromatin in a directional manner.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .