α-Pal/NRF-1 Regulates the Promoter of the Human Integrin-associated Protein/CD47 Gene

Wen Teng Chang, A-Min Huang

研究成果: Article

27 引文 (Scopus)

摘要

Integrin-associated protein (IAP or CD47) is expressed in a variety of tissues, including the nervous system and immune system. To understand how cells control the expression of the IAP gene, we cloned the 5′-proximal region of the human IAP gone and investigated IAP promoter activity by transient transfection. RT-PCR confirmed the expression of IAP transcripts in human neuroblastoma IMR-32 and hepatoma HepG2 cells. Deletion analysis identified a core promoter of the human IAP gene located between nucleotide positions -232 and -12 relative to the translation initiation codon in these two cell lines. Site-directed mutagenesis and gel electrophoretic mobility shift assay identified a α-Pal/NRF-1 binding element within the IAP core promoter. Supershift assays using the α-Pal/NRF-1 antiserum confirmed the binding of this transcription factor on the α-Pal/NRF-1 site. Overexpression of the DNA binding domain of α-Pal/NRF-1 in cells enhanced DNA-α-Pal/NRF-1 binding in vitro. Furthermore, overexpression of full-length α-Pal/NRF-1 significantly enhanced IAP promoter activity while overexpression of dominant-negative mutant reduced promoter activity both in the cultured human cell lines and primary mouse cortical cells. These results revealed that α-Pal/NRF-1 is an essential transcription factor in the regulation of human IAP gene expression.

原文English
頁(從 - 到)14542-14550
頁數9
期刊Journal of Biological Chemistry
279
發行號15
DOIs
出版狀態Published - 2004 四月 9

指紋

Integrins
Assays
Transcription Factors
Genes
Cells
Electrophoretic mobility
Mutagenesis
Immune system
DNA
Neurology
Gene expression
Immune Sera
Proteins
Nucleotides
Gels
Intracisternal A-Particle Genes
Tissue
Gene Expression
Cell Line
Initiator Codon

All Science Journal Classification (ASJC) codes

  • Biochemistry

引用此文

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abstract = "Integrin-associated protein (IAP or CD47) is expressed in a variety of tissues, including the nervous system and immune system. To understand how cells control the expression of the IAP gene, we cloned the 5′-proximal region of the human IAP gone and investigated IAP promoter activity by transient transfection. RT-PCR confirmed the expression of IAP transcripts in human neuroblastoma IMR-32 and hepatoma HepG2 cells. Deletion analysis identified a core promoter of the human IAP gene located between nucleotide positions -232 and -12 relative to the translation initiation codon in these two cell lines. Site-directed mutagenesis and gel electrophoretic mobility shift assay identified a α-Pal/NRF-1 binding element within the IAP core promoter. Supershift assays using the α-Pal/NRF-1 antiserum confirmed the binding of this transcription factor on the α-Pal/NRF-1 site. Overexpression of the DNA binding domain of α-Pal/NRF-1 in cells enhanced DNA-α-Pal/NRF-1 binding in vitro. Furthermore, overexpression of full-length α-Pal/NRF-1 significantly enhanced IAP promoter activity while overexpression of dominant-negative mutant reduced promoter activity both in the cultured human cell lines and primary mouse cortical cells. These results revealed that α-Pal/NRF-1 is an essential transcription factor in the regulation of human IAP gene expression.",
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α-Pal/NRF-1 Regulates the Promoter of the Human Integrin-associated Protein/CD47 Gene. / Chang, Wen Teng; Huang, A-Min.

於: Journal of Biological Chemistry, 卷 279, 編號 15, 09.04.2004, p. 14542-14550.

研究成果: Article

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AB - Integrin-associated protein (IAP or CD47) is expressed in a variety of tissues, including the nervous system and immune system. To understand how cells control the expression of the IAP gene, we cloned the 5′-proximal region of the human IAP gone and investigated IAP promoter activity by transient transfection. RT-PCR confirmed the expression of IAP transcripts in human neuroblastoma IMR-32 and hepatoma HepG2 cells. Deletion analysis identified a core promoter of the human IAP gene located between nucleotide positions -232 and -12 relative to the translation initiation codon in these two cell lines. Site-directed mutagenesis and gel electrophoretic mobility shift assay identified a α-Pal/NRF-1 binding element within the IAP core promoter. Supershift assays using the α-Pal/NRF-1 antiserum confirmed the binding of this transcription factor on the α-Pal/NRF-1 site. Overexpression of the DNA binding domain of α-Pal/NRF-1 in cells enhanced DNA-α-Pal/NRF-1 binding in vitro. Furthermore, overexpression of full-length α-Pal/NRF-1 significantly enhanced IAP promoter activity while overexpression of dominant-negative mutant reduced promoter activity both in the cultured human cell lines and primary mouse cortical cells. These results revealed that α-Pal/NRF-1 is an essential transcription factor in the regulation of human IAP gene expression.

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