Effects of transcription factor GATA-2 on transcriptive regulation of iASPP gene.
10.7534/j.issn.1009-2137.2013.03.003
- Author:
Hai-Yan XING
1
;
Yu-Jiao JIA
;
Ke-Jing TANG
;
Zheng TIAN
;
Yi-Rui CHEN
;
Qing RAO
;
Min WANG
;
Jian-Xiang WANG
Author Information
1. Chinese Academy of Medical Sciences, Tianjin, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Cell Line;
Cercopithecus aethiops;
GATA2 Transcription Factor;
genetics;
Gene Expression Regulation, Leukemic;
Humans;
Intracellular Signaling Peptides and Proteins;
genetics;
K562 Cells;
Repressor Proteins;
genetics;
Transcription, Genetic;
Transcriptional Activation;
Transfection
- From:
Journal of Experimental Hematology
2013;21(3):550-555
- CountryChina
- Language:Chinese
-
Abstract:
iASPP can prompt the cell proliferation and inhibit the apoptosis of many cells. There are putative binding sites of transcription factor GATA-2 upstream of iASPP transcription start site. GATA-2 plays an important role in the proliferation and differentiation of hematopoietic stem cells (HSC) and progenitors. This study was aimed to explore the role of GATA-2 protein in iASPP gene transcription. Firstly, the expression of iASPP and GATA-2 protein in some leukemia cell lines was detected by Western blot. Second, The expressive vector of pCMV5-GATA2 and the luciferase reporter vectors containing possible binding sites of GATA-2 were constructed and co-transfected into HEK293 and CV-1 cells. Then the luciferase activity was assayed by luminometer. Also, ChIP assays were performed to further confirm the specific binding of GATA-2 to iASPP promoter. The results showed that GATA-2 was overexpressed in most cell lines with high level of iASPP. GATA-2 exhibited a significant effect on luciferase activity of reporter gene iASPP and in a dose-dependant manner. The relative luciferase activity was up-regulated to about two-fold of the empty vector control when the transfection dose of pCMV5-GATA2 plasmid was increased to 100 ng. While the effect was more significant in CV-1 cells and showed a 6.7-fold increase. The ChIP assay demonstrated the in vivo specific binding of GATA-2 to iASPP. The binding sites of GATA2 were located between nt -361 ∼ -334 in upstream of iASPP gene transcription start site. It is concluded that transcription factor GATA-2 can bind with the cis-regulatory region of the iASPP promoter and up-regulate iASPP expression.
