Sp1/3 and NF-1 mediate basal transcription of the human P2X1 gene in megakaryoblastic MEG-01 cells

BMC Molecular Biology
Jiangqin Zhao, Steven J Ennion

Abstract

P2X1 receptors play an important role in platelet function as they can induce shape change, granule centralization and are also involved in thrombus formation. As platelets have no nuclei, the level of P2X1 expression depends on transcriptional regulation in megakaryocytes, the platelet precursor cell. Since nothing is known about the molecular mechanisms regulating megakaryocytic P2X1 expression, this study aimed to identify and functionally characterize the P2X1 core promoter utilized in the human megakaryoblastic cell line MEG-01. In order to identify cis-acting elements involved in the transcriptional regulation of P2X1 expression, the ability of 4.7 kb P2X1 upstream sequence to drive luciferase reporter gene expression was tested. Low promoter activity was detected in proliferating MEG-01 cells. This activity increased 20-fold after phorbol-12-myristate-13-acetate (PMA) induced differentiation. A transcription start site was detected 365 bp upstream of the start codon by primer extension. Deletion analysis of reporter constructs indicated a core promoter located within the region -68 to +149 bp that contained two Sp1 sites (named Sp1a and Sp1b) and an NF-1 site. Individual mutations of Sp1b or NF-1 binding sites severely r...Continue Reading

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Citations

Mar 21, 2008·Molecular & Cellular Proteomics : MCP·Yu QuanTao Tao
Mar 14, 2007·Molecular and Cellular Biology·Katrin DrillerAlbrecht E Sippel
Oct 24, 2009·Protein Engineering, Design & Selection : PEDS·Dave E DunstanJoseph Bertolini
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Aug 12, 2015·Purinergic Signalling·Geoffrey Burnstock
Jan 23, 2021·Biochemical and Biophysical Research Communications·Takaomi NozawaKazushi Imai

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Datasets Mentioned

BETA
BX644575

Methods Mentioned

BETA
PMA
ChIP
PCR
transfection
Assay
electrophoresis
immunoprecipitation

Software Mentioned

PROSCAN
TESS
BLAST

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