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

Joeva J. Barrow, Ying Li, Mir Hossain, Suming Huang, Jörg Bungert

Research output: Contribution to journalArticle

8 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)4363-4374
Number of pages12
JournalNucleic acids research
Volume42
Issue number7
DOIs
StatePublished - Apr 2014

Fingerprint

Globins
Zinc Fingers
Genetic Promoter Regions
Locus Control Region
DNA
Upstream Stimulatory Factors
Personnel Selection
Genes
Leukemia, Erythroblastic, Acute
Exonucleases
RNA Polymerase II
Transcription Initiation Site
Transcription Factors
Gene Expression

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

Barrow, Joeva J. ; Li, Ying ; Hossain, Mir ; Huang, Suming ; Bungert, Jörg. / Dissecting the function of the adult β-globin downstream promoter region using an artificial zinc finger DNA-binding domain. In: Nucleic acids research. 2014 ; Vol. 42, No. 7. pp. 4363-4374.
@article{a29f286974f747baa59ad4b4707c1972,
title = "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.",
author = "Barrow, {Joeva J.} and Ying Li and Mir Hossain and Suming Huang and J{\"o}rg Bungert",
year = "2014",
month = "4",
doi = "10.1093/nar/gku107",
language = "English (US)",
volume = "42",
pages = "4363--4374",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "7",

}

Dissecting the function of the adult β-globin downstream promoter region using an artificial zinc finger DNA-binding domain. / Barrow, Joeva J.; Li, Ying; Hossain, Mir; Huang, Suming; Bungert, Jörg.

In: Nucleic acids research, Vol. 42, No. 7, 04.2014, p. 4363-4374.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Dissecting the function of the adult β-globin downstream promoter region using an artificial zinc finger DNA-binding domain

AU - Barrow, Joeva J.

AU - Li, Ying

AU - Hossain, Mir

AU - Huang, Suming

AU - Bungert, Jörg

PY - 2014/4

Y1 - 2014/4

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84899013571&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84899013571&partnerID=8YFLogxK

U2 - 10.1093/nar/gku107

DO - 10.1093/nar/gku107

M3 - Article

C2 - 24497190

AN - SCOPUS:84899013571

VL - 42

SP - 4363

EP - 4374

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 7

ER -