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Stable HeLa Luciferase Reporter Cell Lines Expressing GAL4 Fusion,,,Transactivators


Stable reporter cell lines for signal transduction pathway readout

Lisa Hexdall Li Xu Alan Greener Mary Buchanan John Bauer Chao-Feng Zheng
Stratagene

Stratagene now offers the PathDetect HLR-CREB, HLR-Elk1, and HLR-cJun, cell lines F for studying signal transduction pathways that converge at the transcription factors CREB, Elk1, and c-Jun. These stable HeLa cell lines feature constitutive expression of the GAL4-CREB, GAL4-Elk1, or GAL4-cJun fusion proteins, respectively. They also contain the firefly luciferase gene, which is controlled by a promoter that responds to GAL4 fusions, as does the HLR cell line that contains only the GAL4-responsive luciferase expression plasmid. When activated by phosphorylation, these fusion proteins bind to the promoter and induce luciferase expression. Therefore, luciferase activity in stable cell lines reflects the activation status of the fusion transactivator and, hence, the activation status of corresponding signaling pathways. The effects of extracellular stimuli on signaling pathways that converge at these activators can be studied directly, without transfection. The potential applications of these cell lines include studying signal transduction and transcription mechanisms and assessing protein-protein interactions in mammalian cells. In addition, these cell lines can potentially be adapted for high-throughput drug screening.

Regulation of gene transcription by extracellular signals is pivotal for controlling growth and differentiation. Because the transcription machinery of eukaryotic cells is located inside the nucleus, signals outside the cells need to be transmitted across the cytoplasmic and nuclear membranes as well as the cytoplasm. This process of signal transmission, better known as signal transduction, involve
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