【求助】请教，SOD的转录因子是谁?
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人SOD的转录因子已经调控信号途径现在清楚了吗？多谢！
SOD 有好几种类型,调控信号途肯定也不少.肯定有几个不同的转录因子调节,甚至一个基因都不是一个调空方式.这是一个报道,是否有用.
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Published ahead of print on February 28, 2008, doi:10.1165/rcmb.2007-0378OC
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Articles by Zelko, I. N.
Articles by Folz, R. J.
American Journal of Respiratory Cell and Molecular Biology. Vol. 39, pp. 243-251, 2008
© 2008 American Thoracic Society
DOI: 10.1165/rcmb.2007-0378OC
Transcription Factors Sp1 and Sp3 Regulate Expression of Human Extracellular Superoxide Dismutase in Lung Fibroblasts
Igor N. Zelko1, Michael R. Mueller1 and Rodney J. Folz1
1 Departments of Medicine and Biochemistry and Molecular Biology University of Louisville, Louisville, Kentucky
Correspondence and requests for reprints should be addressed to Rodney J. Folz, M.D., Ph.D., 550 S. Jackson Street/ACB – A3R43, University of Louisville, Louisville, KY. E-mail: rodney.folz@louisville.edu
The molecular mechanisms that govern the transcription of human extracellular superoxide dismutase (EC-SOD), the major extracellular antioxidant enzyme, are largely unknown. To elucidate the mechanisms involved in human EC-SOD gene regulation and expression, we localized multiple transcription start sites to a finite region located 3.9 kb upstream of the ATG initiation codon. Within this segment, we subcloned a 2.7-kb fragment upstream of a luciferase reporter gene; the resulting construct exhibited strong in vivo promoter activity in two lung-derived cell lines. Deletion analysis of the EC-SOD 5'-flanking sequences identified a minimal 0.3-kb region that had strong basal promoter activity. Computer sequence analysis revealed a putative Sp1-like binding site within the EC-SOD proximal promoter region that lacked a TATA-box and showed a high frequency of GC nucleotides. Binding of Sp1 and Sp3 transcription factors to the EC-SOD promoter was confirmed by DNase I footprint analysis, electophoretic mobility shift assay, and competition and supershift assays. Cotransfection of the EC-SOD promoter–luciferase reporter constructs with plasmids encoding Sp1 and Sp3 into Sp-deficient insect SL2 cells showed strong activation of luciferase gene expression. The occupancy of the EC-SOD promoter by Sp1/Sp3 and RNA polymerase II in vivo was determined by chromatin immunoprecipitation assay and correlated well with levels of EC-SOD expression in lung epithelial cells (A549) and pulmonary fibroblasts (MRC5). Collectively, our results demonstrate the important role Sp1 and Sp3 plays in regulating the expression of human EC-SOD in the lung.
Key Words: extracellular superoxide dismutase • promoter • transcription • Sp1 gene family • antioxidant
CLINICAL RELEVANCE
We have identified key regulatory elements for human extracellular superoxide dismutase (EC-SOD), the major antioxidant enzyme in the lung. Since oxidative stress contributes to the development of lung disease, understanding factors that regulate EC-SOD may provide novel therapeutic strategies.
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Published ahead of print on February 28, 2008, doi:10.1165/rcmb.2007-0378OC
This Article
Full Text
Full Text (PDF)
All Versions of this Article:
2007-0378OCv1
39/2/243 most recent
Alert me when this article is cited
Alert me if a correction is posted
Services
Similar articles in this journal
Similar articles in PubMed
Alert me to new issues of the journal
Download to citation manager
Google Scholar
Articles by Zelko, I. N.
Articles by Folz, R. J.
PubMed
PubMed Citation
Articles by Zelko, I. N.
Articles by Folz, R. J.
American Journal of Respiratory Cell and Molecular Biology. Vol. 39, pp. 243-251, 2008
© 2008 American Thoracic Society
DOI: 10.1165/rcmb.2007-0378OC
Transcription Factors Sp1 and Sp3 Regulate Expression of Human Extracellular Superoxide Dismutase in Lung Fibroblasts
Igor N. Zelko1, Michael R. Mueller1 and Rodney J. Folz1
1 Departments of Medicine and Biochemistry and Molecular Biology University of Louisville, Louisville, Kentucky
Correspondence and requests for reprints should be addressed to Rodney J. Folz, M.D., Ph.D., 550 S. Jackson Street/ACB – A3R43, University of Louisville, Louisville, KY. E-mail: rodney.folz@louisville.edu
The molecular mechanisms that govern the transcription of human extracellular superoxide dismutase (EC-SOD), the major extracellular antioxidant enzyme, are largely unknown. To elucidate the mechanisms involved in human EC-SOD gene regulation and expression, we localized multiple transcription start sites to a finite region located 3.9 kb upstream of the ATG initiation codon. Within this segment, we subcloned a 2.7-kb fragment upstream of a luciferase reporter gene; the resulting construct exhibited strong in vivo promoter activity in two lung-derived cell lines. Deletion analysis of the EC-SOD 5'-flanking sequences identified a minimal 0.3-kb region that had strong basal promoter activity. Computer sequence analysis revealed a putative Sp1-like binding site within the EC-SOD proximal promoter region that lacked a TATA-box and showed a high frequency of GC nucleotides. Binding of Sp1 and Sp3 transcription factors to the EC-SOD promoter was confirmed by DNase I footprint analysis, electophoretic mobility shift assay, and competition and supershift assays. Cotransfection of the EC-SOD promoter–luciferase reporter constructs with plasmids encoding Sp1 and Sp3 into Sp-deficient insect SL2 cells showed strong activation of luciferase gene expression. The occupancy of the EC-SOD promoter by Sp1/Sp3 and RNA polymerase II in vivo was determined by chromatin immunoprecipitation assay and correlated well with levels of EC-SOD expression in lung epithelial cells (A549) and pulmonary fibroblasts (MRC5). Collectively, our results demonstrate the important role Sp1 and Sp3 plays in regulating the expression of human EC-SOD in the lung.
Key Words: extracellular superoxide dismutase • promoter • transcription • Sp1 gene family • antioxidant
CLINICAL RELEVANCE
We have identified key regulatory elements for human extracellular superoxide dismutase (EC-SOD), the major antioxidant enzyme in the lung. Since oxidative stress contributes to the development of lung disease, understanding factors that regulate EC-SOD may provide novel therapeutic strategies.
多谢了！
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