Studies on DNA-protein interactions in the upstream regulatory region of the human ɛ-globin gene promoter

The developmental stage-specific silencing of the human ε-globin gene during embryonic life is controlled, in part, by the silencer (-392bp ～-177bp) upstream of this gene. In order to elucidate its role, the nuclear extract from the human fetal liver has been prepared and the interactions between transacting factors and this silencer element have been examined. By using DNaseI footprinting assay, a major protected region from-278bp to-235bp within this silencer element was identified. Furthermore, we found in gel mobility shift assay and Southwestern blotting assay that there were at least four transacting factors (MW ≈ 32, 28, 26 and 22kD) in the nuclear extract isolated from the human fetal liver, which could specifically bind to this region. Our results suggested that these transacting factors might play an important role in silencing the human embryonic ε-globin gene expression at the fetal stage through the interactions with this silencer.

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Trans-acting factors from the human fetal liver binding to the human ɛ-globin gene silencer

Yan

Jiang

Qian

1997

Cell Res

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The 5′-flanking cis-acting elements of the human ɛ-globin gene associates with the nuclear matrix and binds to the nuclear matrix proteins

Yan

Qian

1998

Cell Res

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The nuclear matrix attachment regions(MARs) and the binding nuclear matrix proteins in the 5'-flanking cisacting elements of the human ε-globin gene have been examined. Using in vitro DNA-matrix binding assay, it has been shown that the positive stage-specific regulatory element (ε-PREII,-446bp ~-419bp) upstream of this gene could specifically associate with the nuclear matrix from K562 cells, indicating that ε-PREII may be an erythroidspecific facultative MAR. In gel mobility shift assay and Southwestern blotting assay, an erythroid-specific nuclear matrix protein (ε-NMP k) in K562 cells has been revealed to bind to this positive regulatory element (ε-PREII). Furthermore, we demonstrated that the silencer (-392bp-177bp) upstream of the human ε-globin gene could associate with the nuclear matrices from K562, HEL and Raji cells. In addition, the nuclear matrix proteins prepared from these three cell lines could also bind to this silencer, suggesting that this silencer element might be a constitutive nuclear matrix attachment region (constitutive MAR). Our results demonstrated that the nuclear matrix and nuclear matrix proteins might play an important role in the regulation of the human ε-globin gene expression.

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Studies on DNA-protein interactions in the upstream regulatory region of the human ɛ-globin gene promoter

Cited by 2 publications

References 18 publications

Trans-acting factors from the human fetal liver binding to the human ɛ-globin gene silencer

Trans-acting factors from the human fetal liver binding to the human ɛ-globin gene silencer

The 5′-flanking cis-acting elements of the human ɛ-globin gene associates with the nuclear matrix and binds to the nuclear matrix proteins

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