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PEA3 cooperates with c-Jun in regulation of HER2/neu transcription
Koshi Matsui1, Kazuhito Sugimori, Hiraku Motomura
1Second Department of Surgery, University of Toyama, Toyama 930-0194, Japan.
Abstract:
HER2/neu overexpressing breast tumors exhibit an increase in polyomavirus enhancer activator 3 (PEA3) expression. We examined the relationship between HER2/neu transcriptional activation and PEA3 in cooperation with c-Jun. HER2/neu promoter activity was decreased by deleting PEA3 binding site, and was downregulated when the PEA3 binding site was mutated. PEA3 and c-Jun each weakly enhanced luciferase expression of the HER2/neu promoter. However, the HER2/neu promoter response to PEA3 was considerably enhanced by c-Jun. Thus, we examined the interaction of PEA3 with c-Jun by the two-hybrid system, the transcriptional activity of PEA3 was specifically enhanced by c-Jun. When PEA3, c-Jun and coactivator p300 were cotransfected in MCF7 cells, the transcriptional activity of HER2/neu was increased by up to 20-fold. PEA3 and c-Jun-induced transcription of HER2/neu promoter was repressed by cotransfection of the dominant negative of p300. These results suggest that PEA3 and c-Jun stimulated synergistically the HER2/neu gene transcription with p300.
Insights
Polyomavirus enhancer activator 3 (PEA3) and c-Jun cooperate to enhance HER2/neu gene transcription in breast tumors. This synergistic action, involving the coactivator p300, offers insights into HER2/neu overexpression mechanisms.
Area of Science:
- Molecular Biology
- Oncology
- Gene Regulation
Background:
- HER2/neu overexpression is common in aggressive breast tumors.
- Polyomavirus enhancer activator 3 (PEA3) expression is elevated in these tumors.
- The precise role of PEA3 in HER2/neu transcriptional activation is not fully understood.
Purpose of the Study:
- To investigate the relationship between HER2/neu transcriptional activation and PEA3.
- To explore the cooperative role of PEA3 with c-Jun in regulating HER2/neu.
- To elucidate the involvement of coactivator p300 in this regulatory pathway.
Main Methods:
- Deletion and mutation analysis of the PEA3 binding site on the HER2/neu promoter.
- Luciferase reporter assays to measure promoter activity.
- Two-hybrid system to assess protein-protein interactions.
- Cotransfection experiments in MCF7 cells with PEA3, c-Jun, and p300.
Main Results:
- Disruption of the PEA3 binding site significantly reduced HER2/neu promoter activity.
- PEA3 and c-Jun individually showed weak enhancement, but synergistically increased HER2/neu transcription.
- c-Jun specifically enhanced the transcriptional activity of PEA3.
- Coexpression of PEA3, c-Jun, and p300 led to a 20-fold increase in HER2/neu transcription.
- Dominant-negative p300 repressed PEA3 and c-Jun-mediated HER2/neu transcription.
Conclusions:
- PEA3 and c-Jun act synergistically to stimulate HER2/neu gene transcription.
- The coactivator p300 is essential for this synergistic transcriptional activation.
- These findings provide a molecular mechanism for PEA3's role in HER2/neu overexpression in breast cancer.
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