Sumoylation of p45/NF-E2

Nuclear positioning and transcriptional activation of the mammalian β-like globin gene locus

Yu Chiau Shyu, Tung Liang Lee, Chun Yuan Ting, Shau Ching Wen, Lie Jiau Hsieh, Yueh Chun Li, Jau Lang Hwang, Chyi Chyang Lin, C. K James Shen

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

NF-E2 is a transcription activator for the regulation of a number of erythroid- and megakaryocytic lineage-specific genes. Here we present evidence that the large subunit of mammalian NF-E2, p45, is sumoylated in vivo in human erythroid K562 cells and in mouse fetal liver. By in vitro sumoylation reaction and DNA transfection experiments, we show that the sumoylation occurs at lysine 368 (K368) of human p45/NF-E2. Furthermore, p45 sumoylation enhances the transactivation capability of NF-E2, and this is accompanied by an increase of the NF-E2 DNA binding affinity. More interestingly, we have found that in K562 cells, the β-globin gene loci in the euchromatin regions are predominantly colocalized with the nuclear bodies promyelocytic leukemia protein (PML) oncogenic domains that are enriched with the PML, SUMO-1, RNA polymerase II, and sumoylatable p45/NF-E2. Chromatin immunoprecipitation assays further showed that the intact sumoylation site of p45/NF-E2 is required for its binding to the DNase I-hypersensitive sites of the β-globin locus control region. Finally, we demonstrated by stable transfection assay that only the wild-type p45, but not its mutant form p45 (K368R), could efficiently rescue β-globin gene expression in the p45-null, erythroid cell line CB3. These data together point to a model of mammalian β-like globin gene activation by sumoylated p45/NF-E2 in erythroid cells.

Original languageEnglish
Pages (from-to)10365-10378
Number of pages14
JournalMolecular and Cellular Biology
Volume25
Issue number23
DOIs
Publication statusPublished - Dec 2005
Externally publishedYes

Fingerprint

Sumoylation
Globins
Erythroid Cells
Transcriptional Activation
K562 Cells
Genes
Transfection
Locus Control Region
Euchromatin
Null Lymphocytes
RNA Polymerase II
Chromatin Immunoprecipitation
Deoxyribonuclease I
DNA
Lysine
Gene Expression
Cell Line
Liver
Promyelocytic Leukemia Protein

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Sumoylation of p45/NF-E2 : Nuclear positioning and transcriptional activation of the mammalian β-like globin gene locus. / Shyu, Yu Chiau; Lee, Tung Liang; Ting, Chun Yuan; Wen, Shau Ching; Hsieh, Lie Jiau; Li, Yueh Chun; Hwang, Jau Lang; Lin, Chyi Chyang; Shen, C. K James.

In: Molecular and Cellular Biology, Vol. 25, No. 23, 12.2005, p. 10365-10378.

Research output: Contribution to journalArticle

Shyu, YC, Lee, TL, Ting, CY, Wen, SC, Hsieh, LJ, Li, YC, Hwang, JL, Lin, CC & Shen, CKJ 2005, 'Sumoylation of p45/NF-E2: Nuclear positioning and transcriptional activation of the mammalian β-like globin gene locus', Molecular and Cellular Biology, vol. 25, no. 23, pp. 10365-10378. https://doi.org/10.1128/MCB.25.23.10365-10378.2005
Shyu, Yu Chiau ; Lee, Tung Liang ; Ting, Chun Yuan ; Wen, Shau Ching ; Hsieh, Lie Jiau ; Li, Yueh Chun ; Hwang, Jau Lang ; Lin, Chyi Chyang ; Shen, C. K James. / Sumoylation of p45/NF-E2 : Nuclear positioning and transcriptional activation of the mammalian β-like globin gene locus. In: Molecular and Cellular Biology. 2005 ; Vol. 25, No. 23. pp. 10365-10378.
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AU - Wen, Shau Ching

AU - Hsieh, Lie Jiau

AU - Li, Yueh Chun

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AU - Lin, Chyi Chyang

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