Dissecting the function of the adult β-globin downstream promoter region using an artificial zinc finger DNA-binding domain.

Abstract:

:Developmental stage-specific expression of the β-type globin genes is regulated by many cis- and trans-acting components. The adult β-globin gene contains an E-box located 60 bp downstream of the transcription start site that has been shown to bind transcription factor upstream stimulatory factor (USF) and to contribute to efficient in vitro transcription. We expressed an artificial zinc finger DNA-binding domain (ZF-DBD) targeting this site (+60 ZF-DBD) in murine erythroleukemia cells. Expression of the +60 ZF-DBD reduced the recruitment and elongation of RNA polymerase II (Pol II) at the adult β-globin gene and at the same time increased the binding of Pol II at locus control region (LCR) element HS2, suggesting that Pol II is transferred from the LCR to the globin gene promoters. Expression of the +60 ZF-DBD also reduced the frequency of interactions between the LCR and the adult β-globin promoter. ChIP-exonuclease-sequencing revealed that the +60ZF-DBD was targeted to the adult β-globin downstream promoter and that the binding of the ZF-DBD caused alterations in the association of USF2 containing protein complexes. The data demonstrate that targeting a ZF-DBD to the adult β-globin downstream promoter region interferes with the LCR-mediated recruitment and activity of Pol II.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Barrow JJ,Li Y,Hossain M,Huang S,Bungert J

doi

10.1093/nar/gku107

subject

Has Abstract

pub_date

2014-04-01 00:00:00

pages

4363-74

issue

7

eissn

0305-1048

issn

1362-4962

pii

gku107

journal_volume

42

pub_type

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