Differential disruption of EWS-FLI1 binding by DNA-binding agents.
PLoS One, 2013;8(7):e69714.
Chen C[1], Wonsey DR, Lemieux ME, Kung AL
Affiliations
PMID: 23894528DOI: 10.1371/journal.pone.0069714
Impact factor: 3.752
Abstract
Fusion of the EWS gene to FLI1 produces a fusion oncoprotein that drives an aberrant gene expression program responsible for the development of Ewing sarcoma. We used a homogenous proximity assay to screen for compounds that disrupt the binding of EWS-FLI1 to its cognate DNA targets. A number of DNA-binding chemotherapeutic agents were found to non-specifically disrupt protein binding to DNA. In contrast, actinomycin D was found to preferentially disrupt EWS-FLI1 binding by comparison to p53 binding to their respective cognate DNA targets in vitro. In cell-based assays, low concentrations of actinomycin D preferentially blocked EWS-FLI1 binding to chromatin, and disrupted EWS-FLI1-mediated gene expression. Higher concentrations of actinomycin D globally repressed transcription. These results demonstrate that actinomycin D preferentially disrupts EWS-FLI1 binding to DNA at selected concentrations. Although the window between this preferential effect and global suppression is too narrow to exploit in a therapeutic manner, these results suggest that base-preferences may be exploited to find DNA-binding compounds that preferentially disrupt subclasses of transcription factors.
MeSH terms
Antineoplastic Agents; Cell Line, Tumor; Chromatin; DNA; Dactinomycin; Drug Screening Assays, Antitumor; High-Throughput Screening Assays; Humans; Oncogene Proteins, Fusion; Protein Binding; Proto-Oncogene Protein c-fli-1; RNA-Binding Protein EWS; Sarcoma, Ewing
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