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

Cell Research
Yu-Chiau ShyuChe-Kun James Shen

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 v...Continue Reading

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Mar 12, 2013·Current Opinion in Hematology·Michael R Tallack, Andrew C Perkins
Jul 30, 2010·Cold Spring Harbor Perspectives in Biology·Lyubomira Chakalova, Peter Fraser
May 29, 2010·Genome Research·Michael R TallackAndrew C Perkins
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Jul 3, 2016·Journal of Translational Medicine·Yu-Chiau ShyuTing-Shuo Huang
Nov 14, 2020·International Journal of Molecular Sciences·Chun-Hao HungChe-Kun James Shen
Aug 8, 2021·International Journal of Molecular Sciences·Chun-Hao HungChe-Kun James Shen

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