Volume 12, No 1, Mar 2002

ISSN: 1001-0602 
EISSN: 1748-7838 2018 
impact factor 17.848* 
(Clarivate Analytics, 2019)

Volume 12 Issue 1, March 2002: 79-82

COMMENTARY

Identification of a NF-κB site in the negative regulatory element(ε-NRAII) of human ε-globin gene and its binding protein NF-κB p50 in the nuclei of K562 cells

Chun Hui HOU, Jian HUANG, Ruo Lan QIAN*

Group of Globin Gene Expression and Regulation, State Key Labortory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China Correspondence:

he developmental control of the human e-globin
gene expression is mediated by transcription regulatory elements in the 5'
flanking DNA of this gene. Sequence analysis has revealed a DNA motif (GGGGAATTTGCT)
similar to NF-kB consensus sequence resides in the
negative regulatory element (-3028bp ~ -2902bp,
termed e-NRAII) 5' to the cap site of this gene. NRF
DNA fragment (-3010bp ~ -2986bp) containing the NF-kB
motif similar sequence was synthesized and used in electrophoresis mobility
shift assay (EMSA) and competitive analysis. Data showed that a protein factor
from nuclear extracts of K562 cells specifically interacted with NRF DNA
fragment. The synthetic NF DNA fragment (containing NF-kB
consensus sequence) could competed for the protein binding, but MNF DNA fragment
(mutated NF-kB motif) could not, suggesting that the
binding protein is a member of NF-kB/Rel family.
Western blot assay demonstrated that the molecular weight of NF-kB
protein in the nuclei of K562 cells is 50ku. We suggested that NF-kB
p50 may play an important role in the regulation of human e-globin
gene expression.

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