Fluorescence-Based Functional Assay for Wnt/β-Catenin Signaling Activity
| Autor: | L. Zhou, N. An, W. Jiang, R. Haydon, H. Cheng, Q. Zhou, B. Breyer, T. Feng, T.-C. He |
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| Jazyk: | angličtina |
| Rok vydání: | 2002 |
| Předmět: | |
| Zdroj: | BioTechniques, Vol 33, Iss 5, Pp 1126-1135 (2002) |
| Druh dokumentu: | article |
| ISSN: | 1940-9818 0736-6205 |
| DOI: | 10.2144/02335dd07 |
| Popis: | Aberrant activation of β-catenin signaling has been implicated in the development of human cancers. As a Wnt signal transducer, β-catenin forms a complex with the lymphocyte enhancer-binding factor/T cell factor transcription factor and activates downstream targets that promote cell proliferation. Here we developed a Wnt-dependent β-catenin-mediated heterologous transactivation system, which consisted of a chimeric transcription factor constructed by fusing the GAL4 DNA-binding domain with the full-length β-catenin, and a GAL4-responsive reporter expressing GFP. The chimeric transcription factor was highly unstable and exerted no detectable transactivating effect on the GAL4-responsive reporter. However, lithium and Wnt1 significantly stabilized this chimeric transactivator, indicating that this transactivation system is regulated by β-catenin in a Wnt-responsive fashion. Thus, this transactivation system could be used as a functional reporter to identify potential upstream factors that deregulate β-catenin signaling during tumorigenesis, as well as to screen for potential anti-cancer agents that specifically inhibit β-catenin signaling in human tumors. |
| Databáze: | Directory of Open Access Journals |
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