Effects of transcription factor GATA-2 on transcriptive regulation of iASPP gene / 中国实验血液学杂志
Journal of Experimental Hematology
; (6): 550-555, 2013.
Article
in Zh
| WPRIM
| ID: wpr-332738
Responsible library:
WPRO
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.
Full text:
1
Index:
WPRIM
Main subject:
Repressor Proteins
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Transcription, Genetic
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Transfection
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Gene Expression Regulation, Leukemic
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Transcriptional Activation
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Cell Line
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Chlorocebus aethiops
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K562 Cells
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Intracellular Signaling Peptides and Proteins
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GATA2 Transcription Factor
Limits:
Animals
/
Humans
Language:
Zh
Journal:
Journal of Experimental Hematology
Year:
2013
Type:
Article