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Response elements of the androgen-regulated C3 gene
J A Tan1, K B Marschke, K C Ho
1Laboratories for Reproductive Biology, University of North Carolina, Chapel Hill 27599.
The Journal of Biological Chemistry
|March 5, 1992
Summary
Researchers identified androgen response elements (AREs) within the C3 subunit gene. These elements, found in the intron and 5'-flanking regions, are crucial for androgen-dependent gene regulation.
Area of Science:
- Molecular Biology
- Genetics
- Endocrinology
Background:
- The C3 subunit gene is regulated by androgens.
- Potential transcription control sequences, resembling glucocorticoid (GRE) and progesterone (PRE) response elements, were identified in the C3 gene's regulatory regions.
Purpose of the Study:
- To investigate whether GRE/PRE-like sequences in the C3 gene function as androgen response elements (AREs).
- To characterize the androgen-dependent transcriptional regulation of the C3 gene.
Main Methods:
- DNA binding assays (mobility shift assay) to detect androgen receptor (AR) binding.
- Reporter gene assays (cotransfection with ptkCAT) to measure transcriptional activation.
- Analysis of C3 gene fragments (5'-flanking and first intron) and synthetic elements cloned into a reporter vector.
Main Results:
- Specific DNA binding of the AR and androgen-dependent transcription activation were observed.
- A 0.5-kbp fragment from the first intron showed significant androgen-dependent enhancement of transcription (16 +/- 4-fold).
- Element C within the intron demonstrated stronger ARE activity than elements B or A, and its activity was comparable to a potent GRE from the mouse mammary tumor virus gene.
Conclusions:
- The C3 gene contains functional androgen response elements (AREs) in its intron and 5'-flanking regions.
- The intron region appears to contain additional regulatory sequences that potentiate ARE activity.
- These findings elucidate the molecular mechanisms of androgen-mediated C3 gene regulation.