期刊
EXPERIMENTAL CELL RESEARCH
卷 315, 期 16, 页码 2690-2704出版社
ELSEVIER INC
DOI: 10.1016/j.yexcr.2009.06.021
关键词
Writ signaling; TCF; Chromatin immunoprecipitation; Central nervous system
资金
- Academy of Finland [107406, 206038]
- Sigrid Juselius Foundation
- European Union [LSHG-CT-2004-005085]
- Academy of Finland (AKA) [107406, 107406] Funding Source: Academy of Finland (AKA)
Writ proteins are important regulators of embryonic development, and dysregulated Writ signalling is involved in the oncogenesis of several human cancers. Our knowledge of the downstream target genes is limited, however. We used a chromatin immunoprecipitation-based assay to isolate and characterize the actual gene segments through which Writ-activatable transcription factors, TCFs, regulate transcription and an Affymetrix microarray analysis to study the global transcriptional response to the Wnt3a ligand. The anti-beta-catenin immunoprecipitation of DNA-protein complexes from mouse NIH3T3 fibroblasts expressing a fusion Protein of beta-catenin and TCF7 resulted in the identification of 92 genes as Putative TCF targets. GeneChip assays of gene expression performed on NIH3T3 cells and the rat pheochromocytoma cell line PC12 revealed 355 genes in NIH3T3 and 129 genes in the PC12 cells with marked changes in expression after Wnt3a stimulus. Only 2 Wnt-regulated genes were shared by both cell lines. Surprisingly, Disabled-2 was the only gene identified by the chromatin immunoprecipitation approach that displayed a marked change in expression in the GeneChip assay. Taken together, our, approaches give an insight into the complex context-dependent nature of Writ pathway transcriptional responses and identify Disabled-2 as a potential new direct target for Writ signalling. (C) 2009 Elsevier Inc. All rights reserved.
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