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Published on: May 23, 2014
Identification of c-Jun as bcl-2 transcription factor in human uterine endometrium
Insights
Biomolecular interaction (BIA) analysis revealed c-Jun protein binds to bcl-2 promoter DNA in human endometrial glandular cells during proliferation. This DNA-protein interaction, unlike immunohistochemistry (IHC) findings, suggests c-Jun regulates bcl-2 transcription for cell growth.
Area of Science:
- Molecular biology
- Cell biology
- Genetics
Background:
- The transcription factor c-Jun plays a role in cellular signaling and proliferation.
- The bcl-2 gene is crucial for cell survival and is often dysregulated in cancer.
- Understanding the regulation of bcl-2 by transcription factors like c-Jun is important for endometrial biology.
Purpose of the Study:
- To investigate the interaction between c-Jun protein and AP-1 motifs in the bcl-2 promoter using biomolecular interaction analysis.
- To compare the results of biomolecular interaction analysis with immunohistochemistry (IHC) for c-Jun expression in the human uterine endometrium.
- To elucidate the role of c-Jun in regulating bcl-2 transcription in endometrial cells.
Main Methods:
- Application of the biomolecular interaction (BIA) technique using a Biacore 2000 system to detect protein-DNA binding.
- Specific binding assay for c-Jun and AP-1 motifs from the bcl-2 promoter.
- Immunohistochemistry (IHC) for c-Jun expression in human uterine endometrial tissue.
Main Results:
- Immunohistochemistry showed intense c-Jun immunoreactivity in glandular cells during the proliferative phase and in stromal cells throughout the menstrual cycle.
- Biomolecular interaction analysis detected specific binding of c-Jun to bcl-2 promoter AP-1 motifs exclusively in nuclear extracts of glandular cells during the proliferative phase.
- No c-Jun binding to bcl-2 promoter DNA was detected in stromal cells using the BIA technique.
Conclusions:
- c-Jun enhances bcl-2 transcription levels in human endometrial glandular cells during the proliferative phase.
- The interaction between c-Jun and the bcl-2 promoter contributes to the survival and proliferation of glandular cells.
- Real-time biomolecular interaction analysis, combined with IHC, offers a novel method for studying transcription factor-DNA binding.
Abstract:
We describe the application of the biomolecular interaction (BIA) technique to detection of the interaction between protein (e.g., c-Jun) and DNA (e.g., two AP-1 motifs from bcl-2 promoter), compared with immunohistochemistry (IHC) of c-Jun. The specific binding assay for the interaction of c-Jun and activating protein-1 (AP-1) motifs was performed using a Biacore 2000 system. Intense immunoreactivity of c-Jun in glandular cells of the human uterine endometrium was observed in the proliferative phase, while c-Jun in stromal cells was expressed throughout the menstrual cycle. In contrast to the IHC of c-Jun, the specific binding of c-Jun to two separate AP-1 motifs in the bcl-2 promoter region was detected only in nuclear extracts of glandular cells, but not in stromal cells, during the proliferative phase. These results indicate that, while transmitting various signals, c-Jun enhances the transcription level of bcl-2, which in turn keeps glandular cells alive and proliferating in normal human endometrium during the proliferative phase. Moreover, the method involving real-time biomolecular interactions such as DNA-protein binding is novel for the study of transcription factors when combined with IHC.
