Fluorescence-based functional assay for Wnt/beta-catenin signaling activity

Aberrant activation of beta-catenin signaling has been implicated in the development of human cancers. As a Wnt signal transducer, beta-catenin forms a complex with the lymphocyte enhancer-binding factor/T cell factor transcription factor and activates downstream targets that promote cell proliferat...

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Publié dans:BioTechniques. - 1991. - 33(2002), 5 vom: 21. Nov., Seite 1126-8, 1130, 1132 passim
Auteur principal: Zhou, L (Auteur)
Autres auteurs: An, N, Jiang, W, Haydon, R, Cheng, H, Zhou, Q, Breyer, B, Feng, T, He, T-C
Format: Article
Langue:English
Publié: 2002
Accès à la collection:BioTechniques
Sujets:Research Support, Non-U.S. Gov't Technical Report Journal Article CTNNB1 protein, human CTNNB1 protein, mouse Culture Media, Conditioned Cytoskeletal Proteins DNA-Binding Proteins GAL4 protein, S cerevisiae Luminescent Proteins plus... Neoplasm Proteins Proto-Oncogene Proteins Recombinant Fusion Proteins Saccharomyces cerevisiae Proteins Trans-Activators Transcription Factors WNT1 protein, human Wnt Proteins Wnt1 Protein Wnt1 protein, mouse Zebrafish Proteins beta Catenin Green Fluorescent Proteins 147336-22-9 Luciferases EC 1.13.12.-
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Résumé:Aberrant activation of beta-catenin signaling has been implicated in the development of human cancers. As a Wnt signal transducer, beta-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 beta-catenin-mediated heterologous transactivation system, which consisted of a chimeric transcription factor constructed by fusing the GAL4 DNA-binding domain with the full-length beta-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 beta-catenin in a Wnt-responsive fashion. Thus, this transactivation system could be used as a functional reporter to identify potential upstream factors that deregulate beta-catenin signaling during tumorigenesis, as well as to screen for potential anti-cancer agents that specifically inhibit beta-catenin signaling in human tumors
Description:Date Completed 17.06.2003
Date Revised 28.09.2018
published: Print
Citation Status MEDLINE
ISSN:1940-9818