[具有-173 T→C突变的人类Aγ-珠蛋白基因启动子的功能分析]

[Functional analysis of human A gamma-globin gene promoter with-173 T-->C mutation].

作者信息

Huang X, Zhang J, Yang X, Wang Y, Zhao H

机构信息

National Laboratory of Medical Molecular Biology, Institute of Basic of Medical Sciences, CAMS and PUMC, Beijing 100005.

出版信息

Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 1999 Aug;21(4):252-6.

PMID:12567445

Abstract

OBJECTIVE

To investigate the effect of the human A gamma-globin gene-173 T-->C mutation on the binding of transacting factors to the promoter and the activity of the promoter.

METHODS

Electrophoretic mobility shift assay and transient transfection assay were used in this study.

RESULTS

The A gamma-globin gene-173 T-->C mutation decreased the affinity of GATA-1 to the mutant promoter fragment (-201(-)-158) 96% (P < 0.01) and the binding of Oct-1 to the same DNA fragment 55% (P < 0.05). The activity of the mutant promoter was 2-fold (P < 0.05) as strong as that of the normal one in MELGM979 cells. The mutant and the normal promoters exhibited basically same activities in K562 and Hela cells.

CONCLUSIONS

The -173 T-->C mutation decreased dramatically the binding of GATA-1, implicating that GA-TA-1 may act as a negative regulator of A gamma-globin gene in adults. The -173 T-->C mutation may enhance the activity of A gamma-globin gene promoter in the adult erythroid cell environment.

摘要

目的

研究人类Aγ-珠蛋白基因-173 T→C突变对反式作用因子与启动子结合及启动子活性的影响。

方法

本研究采用电泳迁移率变动分析和瞬时转染分析。

结果

Aγ-珠蛋白基因-173 T→C突变使GATA-1与突变型启动子片段（-201（-）-158）的亲和力降低96%（P<0.01），使Oct-1与同一DNA片段的结合降低55%（P<0.05）。在MELGM979细胞中，突变型启动子的活性是正常启动子的2倍（P<0.05）。突变型和正常启动子在K562和Hela细胞中表现出基本相同的活性。