Chromatin-binding in vivo of the erythroid kruppel-like factor, EKLF, in the murine globin loci.

Abstract:

:EKLF is an erythroid-specific, zinc finger-containing transcription factor essential for the activation of the mammalian beta globin gene in erythroid cells of definitive lineage. We have prepared a polyclonal anti-mouse EKLF antibody suitable for Western blotting and immunoprecipitation (IP) qualities, and used it to define the expression patterns of the EKLF protein during mouse erythroid development. We have also used this antibody for the chromatin-immunoprecipitation (ChIP) assay. EKLF was found to bind in vivo at both the mouse beta-major-globin promoter and the HS2 site of beta-LCR in the mouse erythroleukemia cells (MEL) in a DMSO-inducible manner. The DMSO-induced bindings of EKLF as well as three other proteins, namely, RNA polymerase II, acetylated histone H3, and methylated histone H3, were not abolished but significantly lowered in CB3, a MEL-derived cell line with null-expression of p45/NF-E2, an erythroid-enriched factor needed for activation of the mammalian globin loci. Interestingly, binding of EKLF in vivo was also detected in the mouse alpha-like globin locus, at the adult alpha globin promoter and its far upstream regulatory element alpha-MRE (HS26). This study provides direct evidence for EKLF-binding in vivo at the major regulatory elements of the mouse beta-like globin gene clusters the data also have interesting implications with respect to the role of EKLF-chromatin interaction in mammalian globin gene regulation.

journal_name

Cell Res

journal_title

Cell research

authors

Shyu YC,Wen SC,Lee TL,Chen X,Hsu CT,Chen H,Chen RL,Hwang JL,Shen CK

doi

10.1038/sj.cr.7310045

subject

Has Abstract

pub_date

2006-04-01 00:00:00

pages

347-55

issue

4

eissn

1001-0602

issn

1748-7838

pii

7310045

journal_volume

16

pub_type

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