title: Differential disruption of EWS-FLI1 binding by DNA-binding agents.
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.
authors.affiliation: Department of Pediatric Oncology, Dana-Farber Cancer Institute and Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Changmin
authors.initials: C
authors.lastname: Chen
authors.forename: Diane R
authors.initials: DR
authors.lastname: Wonsey
authors.forename: Madeleine E
authors.initials: ME
authors.lastname: Lemieux
authors.forename: Andrew L
authors.initials: AL
authors.lastname: Kung
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology* / Q000627:therapeutic use; D045744:Cell Line, Tumor; D002843:Chromatin / Q000378:metabolism; D004247:DNA / Q000378:metabolism*; D003609:Dactinomycin / Q000494:pharmacology; D004354:Drug Screening Assays, Antitumor; D057166:High-Throughput Screening Assays; D006801:Humans; D015514:Oncogene Proteins, Fusion / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D051789:Proto-Oncogene Protein c-fli-1 / Q000378:metabolism*; D034802:RNA-Binding Protein EWS / Q000378:metabolism*; D012512:Sarcoma, Ewing / Q000188:drug therapy / Q000473:pathology
chemical_list: D000970:Antineoplastic Agents; D002843:Chromatin; C106031:EWS-FLI fusion protein; D015514:Oncogene Proteins, Fusion; D051789:Proto-Oncogene Protein c-fli-1; D034802:RNA-Binding Protein EWS; D003609:Dactinomycin; D004247:DNA
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