Abstract
We have characterized three cis-acting elements of the human CYP11A1 gene. A proximal cAMP-responsive sequence (P-CRS) functioned in both adrenal Y1 and placental JEG-3 cells. An upstream cAMP-responsive sequence (U-CRS) and an enhancer, localized by transfections of deleted gene segments linked to a reporter gene to bases -1621 to -1503 and -1931 to -1822, respectively, functioned in Y1 but not JEG-3 cells. Both regions bind proteins only from Y1 cells as identified by footprinting analysis. U-CRS contains the TCAAGGTCA sequence that binds the nuclear receptor family of proteins. The cAMP- dependent transcription mediated by U-CRS, but not by P-CRS, was abolished in a cell line deficient in cAMP-dependent protein kinase. Therefore, P-CRS and U-CRS use different effectors to mediate cAMP response. Gel mobility shift, competition, and antibody supershift experiments showed that nucleotides - 117 to -94, which contributed to P-CRS activity in transfection experiments, bound weakly to Sp1-like proteins. This feature is shared by many proximal regulatory elements of steroidogenic genes. Therefore, steroidogenic genes could be coordinately regulated through common regulatory elements such as P- CRS, U-CRS, and cell type-selective enhancers.
| Original language | English |
|---|---|
| Pages (from-to) | 6362-6369 |
| Number of pages | 8 |
| Journal | Journal of Biological Chemistry |
| Volume | 269 |
| Issue number | 9 |
| Publication status | Published - Mar 4 1994 |
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All Science Journal Classification (ASJC) codes
- Biochemistry
- Molecular Biology
- Cell Biology
Cite this
Actions of two different cAMP-responsive sequences and an enhancer of the human CYP11A1 (P450scc) gene in adrenal Y1 and placental JEG-3 cells. / Guo, Ing Cherng; Tsai, Huei Man; Chung, Bon Chu.
In: Journal of Biological Chemistry, Vol. 269, No. 9, 04.03.1994, p. 6362-6369.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Actions of two different cAMP-responsive sequences and an enhancer of the human CYP11A1 (P450scc) gene in adrenal Y1 and placental JEG-3 cells
AU - Guo, Ing Cherng
AU - Tsai, Huei Man
AU - Chung, Bon Chu
PY - 1994/3/4
Y1 - 1994/3/4
N2 - We have characterized three cis-acting elements of the human CYP11A1 gene. A proximal cAMP-responsive sequence (P-CRS) functioned in both adrenal Y1 and placental JEG-3 cells. An upstream cAMP-responsive sequence (U-CRS) and an enhancer, localized by transfections of deleted gene segments linked to a reporter gene to bases -1621 to -1503 and -1931 to -1822, respectively, functioned in Y1 but not JEG-3 cells. Both regions bind proteins only from Y1 cells as identified by footprinting analysis. U-CRS contains the TCAAGGTCA sequence that binds the nuclear receptor family of proteins. The cAMP- dependent transcription mediated by U-CRS, but not by P-CRS, was abolished in a cell line deficient in cAMP-dependent protein kinase. Therefore, P-CRS and U-CRS use different effectors to mediate cAMP response. Gel mobility shift, competition, and antibody supershift experiments showed that nucleotides - 117 to -94, which contributed to P-CRS activity in transfection experiments, bound weakly to Sp1-like proteins. This feature is shared by many proximal regulatory elements of steroidogenic genes. Therefore, steroidogenic genes could be coordinately regulated through common regulatory elements such as P- CRS, U-CRS, and cell type-selective enhancers.
AB - We have characterized three cis-acting elements of the human CYP11A1 gene. A proximal cAMP-responsive sequence (P-CRS) functioned in both adrenal Y1 and placental JEG-3 cells. An upstream cAMP-responsive sequence (U-CRS) and an enhancer, localized by transfections of deleted gene segments linked to a reporter gene to bases -1621 to -1503 and -1931 to -1822, respectively, functioned in Y1 but not JEG-3 cells. Both regions bind proteins only from Y1 cells as identified by footprinting analysis. U-CRS contains the TCAAGGTCA sequence that binds the nuclear receptor family of proteins. The cAMP- dependent transcription mediated by U-CRS, but not by P-CRS, was abolished in a cell line deficient in cAMP-dependent protein kinase. Therefore, P-CRS and U-CRS use different effectors to mediate cAMP response. Gel mobility shift, competition, and antibody supershift experiments showed that nucleotides - 117 to -94, which contributed to P-CRS activity in transfection experiments, bound weakly to Sp1-like proteins. This feature is shared by many proximal regulatory elements of steroidogenic genes. Therefore, steroidogenic genes could be coordinately regulated through common regulatory elements such as P- CRS, U-CRS, and cell type-selective enhancers.
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M3 - Article
VL - 269
SP - 6362
EP - 6369
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 9
ER -