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

Yu Chiau Shyu, Shau Ching Wen, Tung Liang Lee, Xin Chen, Chia Tse Hsu, Hsin Chen, Ruei Lin Chen, Jau Lang Hwang, Che Kun James Shen

研究成果: 雜誌貢獻文章

17 引文 (Scopus)

摘要

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.

原文英語
頁(從 - 到)347-355
頁數9
期刊Cell Research
16
發行號4
DOIs
出版狀態已發佈 - 四月 2006
對外發佈Yes

指紋

Globins
Chromatin
beta-Globins
alpha-Globins
Leukemia, Erythroblastic, Acute
Dimethyl Sulfoxide
Histones
Erythroid Cells
erythroid Kruppel-like factor
Antibodies
RNA Polymerase II
Chromatin Immunoprecipitation
Zinc Fingers
Cell Lineage
Multigene Family
Immunoprecipitation
Genes
Proteins
Transcription Factors
Western Blotting

ASJC Scopus subject areas

  • Cell Biology

引用此文

Shyu, Y. C., Wen, S. C., Lee, T. L., Chen, X., Hsu, C. T., Chen, H., ... Shen, C. K. J. (2006). Chromatin-binding in vivo of the erythroid kruppel-like factor, EKLF, in the murine globin loci. Cell Research, 16(4), 347-355. https://doi.org/10.1038/sj.cr.7310045

Chromatin-binding in vivo of the erythroid kruppel-like factor, EKLF, in the murine globin loci. / Shyu, Yu Chiau; Wen, Shau Ching; Lee, Tung Liang; Chen, Xin; Hsu, Chia Tse; Chen, Hsin; Chen, Ruei Lin; Hwang, Jau Lang; Shen, Che Kun James.

於: Cell Research, 卷 16, 編號 4, 04.2006, p. 347-355.

研究成果: 雜誌貢獻文章

Shyu, YC, Wen, SC, Lee, TL, Chen, X, Hsu, CT, Chen, H, Chen, RL, Hwang, JL & Shen, CKJ 2006, 'Chromatin-binding in vivo of the erythroid kruppel-like factor, EKLF, in the murine globin loci', Cell Research, 卷 16, 編號 4, 頁 347-355. https://doi.org/10.1038/sj.cr.7310045
Shyu, Yu Chiau ; Wen, Shau Ching ; Lee, Tung Liang ; Chen, Xin ; Hsu, Chia Tse ; Chen, Hsin ; Chen, Ruei Lin ; Hwang, Jau Lang ; Shen, Che Kun James. / Chromatin-binding in vivo of the erythroid kruppel-like factor, EKLF, in the murine globin loci. 於: Cell Research. 2006 ; 卷 16, 編號 4. 頁 347-355.
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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.",
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AU - Shyu, Yu Chiau

AU - Wen, Shau Ching

AU - Lee, Tung Liang

AU - Chen, Xin

AU - Hsu, Chia Tse

AU - Chen, Hsin

AU - Chen, Ruei Lin

AU - Hwang, Jau Lang

AU - Shen, Che Kun James

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N2 - 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.

AB - 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.

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