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DNA binding proteins that amplify surfactant protein B gene expression: isolation and characterization
1Department of Pathology and Cell Biology, Jefferson Medical College, Philadelphia, Pennsylvania 19107.
Biochemical and Biophysical Research Communications
|March 8, 1995
Summary
Two novel proteins, A2 and B, bind the surfactant protein B (SP-B) promoter, influencing its expression. Protein B enhances SP-B activity, particularly with dexamethasone, suggesting new gene activators for SP-B expression.
Area of Science:
- Molecular Biology
- Gene Regulation
Background:
- Surfactant protein B (SP-B) is crucial for lung function.
- The regulation of SP-B gene expression involves complex protein-DNA interactions.
Purpose of the Study:
- To identify and characterize novel proteins that regulate surfactant protein B (SP-B) gene expression.
- To investigate the role of these proteins in basal and glucocorticoid-induced SP-B promoter activity.
Main Methods:
- Proteins A2 and B were identified, and their cDNAs were cloned and sequenced.
- Electrophoretic mobility shift assays (EMSAs) or similar techniques were used to determine DNA binding.
- Reporter gene assays (luciferase) in H441 cells were performed to assess promoter activity with and without protein expression and dexamethasone treatment.
Main Results:
- Two novel proteins, A2 and B, were identified as binding to the SP-B promoter at an NF1 site (-184 to -198).
- Expression of A2 and B individually resulted in significant increases in SP-B promoter activity.
- Dexamethasone treatment further enhanced the effect of protein B on SP-B promoter activity.
Conclusions:
- Proteins A2 and B are novel activators of the SP-B gene.
- Basal SP-B expression and its response to glucocorticoids involve previously unidentified gene activators.
- These findings provide new insights into the regulatory mechanisms of SP-B.