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Multifactor complex containing B element binding factor, BBF, and repressors regulate the human alpha 1(III) collagen
Hideaki Sumiyoshi1, Noritaka Matsuo, Toshitaka Shin
1Department of Anatomy, Biology and Medicine, Oita University, Faculty of Medicine, Oita 879-5593, Japan.
Acta Medica Okayama
|July 14, 2006
Summary
Cell type influences how the type III collagen gene (COL3A1) promoter functions. Specific nuclear factors binding to the promoter change, altering gene expression and cell characteristics, particularly in RD cells.
Area of Science:
- Biochemistry
- Molecular Biology
- Genetics
Background:
- Type III collagen is crucial in fetal skin and blood vessels.
- Previous studies characterized the proximal promoter of the human alpha1(III) collagen gene (COL3A1).
Purpose of the Study:
- To further analyze the COL3A1 promoter using new cell lines.
- To investigate cell-type-specific binding of nuclear factors to the COL3A1 promoter.
- To examine the impact of promoter activity on cell characteristics.
Main Methods:
- Luciferase assay
- Electrophoretic mobility shift assays (EMSA)
- DNase footprinting assay
- Stable transfection with GFP-linked COL3A1 promoter fragment
Main Results:
- Two types of multifactor complexes bind to the COL3A1 promoter region (-80 to -58) in a cell-type-dependent manner.
- Transfected RD cells showed altered promoter expression and cell morphology, mimicking A204 cells.
- Nuclear factors in these altered RD cell clones resembled those in A204 cells.
Conclusions:
- The COL3A1 promoter activity is regulated by cell-specific nuclear factor interactions.
- Cellular context can induce significant changes in promoter function and cell phenotype.
- These findings offer insights into the molecular mechanisms controlling COL3A1 gene expression.