Identification of target genes for EWS/ATF-1 chimeric transcription factor
| Autor: | Yukari Yamazaki, Hiroshi Kuroda, Takuro Nakamura, Miki Jishage, Takashi Fujino |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Adult
Male Cancer Research viruses Recombinant Fusion Proteins genetic processes Molecular Sequence Data Down-Regulation Apoptosis Biology environment and public health Gene expression Genetics Tumor Cells Cultured Humans Binding site Molecular Biology Transcription factor Gene Activating Transcription Factor 1 Base Sequence fungi Promoter DNA Molecular biology Fusion protein Up-Regulation DNA-Binding Proteins Regulatory sequence RNA-Binding Protein EWS Chromatin immunoprecipitation Transcription Factors |
| Zdroj: | Oncogene. 22(1) |
| ISSN: | 0950-9232 |
| Popis: | Chromatin immunoprecipitation is a useful technique to detect in vivo direct interaction between any transcription factor and its binding site on genomic DNA. We applied this skill to identify the direct target gene for EWS/ATF-1 by coupling with a GFP reporter assay. This novel approach isolated 62 of cloned DNA fragments responding upon EWS/ATF-1 expression and 16 of 62 clones included putative ATF-1 binding sites. Further analysis revealed that six of the cloned fragments included possible regulatory regions of ATM, GPP34, ARNT2, NKX6.1, NYD-SP28 and POSH. Most of these clones upregulated reporter activity by overexpression of EWS/ATF-1, suggesting that putative ATF-1 binding sites in these clones are functional elements for ATF-1 in vivo. Consistently, endogenous expression of these genes was upregulated by EWS/ATF-1. Interestingly, the clone containing the promoter region of POSH, which is known to be a strong inducer of apoptosis, repressed reporter activity by overexpression of EWS/ATF-1. Correspondingly, EWS/ATF-1 expression decreased endogenous POSH expression, suggesting that six isolated genes may be involved in direct regulation by EWS/ATF-1. Moreover, induction of POSH brought apoptotic cell death to KAS, the clear cell sarcoma (CCS) cell line, suggesting that repressed expression of POSH in CCS may be relevant to the normal signaling pathway in apoptosis. |
| Databáze: | OpenAIRE |
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