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Functional analysis of the transcriptional coactivators p300 and CBP using ribozyme
H Kawasaki1, K Taira, K K Yokoyama
1Tsukuba Life Science Center, The Institute of Physical and Chemical Research (RIKEN), Tsukuba Science City, Japan.
Abstract:
p300 and CERB binding protein (CBP) are transcriptional coactivator which participate many signal transduction pathways via interaction with DNA binding proteins. In this study, we investigated functions of p300 and CBP during the retinoic acid (RA) induced F9 cell differentiation using hammerhead ribozymes. In wild type F9 cells, induction of c-jun gene was observed at 18 h after treatment with RA. Interestingly, RA-dependent expression of c-jun gene was not induced in cells expressing the p300-directed ribozyme. By contrast, the CBP-directed ribozyme did not affect expression of the c-jun gene. These results suggest that p300 is critical for the induction of c-jun gene during RA-induced differentiation.
Insights
p300, a transcriptional coactivator, is crucial for retinoic acid-induced c-jun gene expression in F9 cell differentiation. CBP does not play a significant role in this specific pathway.
Area of Science:
- Molecular Biology
- Cellular Differentiation
- Gene Regulation
Background:
- p300 and CREB Binding Protein (CBP) are transcriptional coactivators involved in numerous signaling pathways.
- These coactivators interact with DNA-binding proteins to regulate gene expression.
Purpose of the Study:
- To investigate the specific roles of p300 and CBP in retinoic acid (RA)-induced differentiation of F9 cells.
- To determine the involvement of p300 and CBP in the induction of the c-jun gene during this process.
Main Methods:
- Utilized hammerhead ribozymes to specifically inhibit the function of p300 and CBP in F9 cells.
- Treated cells with retinoic acid (RA) to induce differentiation.
- Monitored the expression of the c-jun gene using quantitative methods.
Main Results:
- In wild-type F9 cells, RA treatment led to c-jun gene induction within 18 hours.
- Inhibition of p300 using a directed ribozyme abolished RA-dependent c-jun gene expression.
- Inhibition of CBP did not affect the expression of the c-jun gene.
Conclusions:
- p300 plays a critical role in the induction of the c-jun gene during RA-induced F9 cell differentiation.
- CBP does not appear to be essential for c-jun gene induction in this specific context.