Differential Disruption of EWS-FLI1 Binding by DNA-Binding Agents

被引:6
作者
Chen, Changmin [1 ,2 ]
Wonsey, Diane R. [1 ,2 ]
Lemieux, Madeleine E. [1 ,2 ]
Kung, Andrew L. [3 ]
机构
[1] Harvard Univ, Sch Med, Dana Farber Canc Inst, Dept Pediat Oncol, Boston, MA 02115 USA
[2] Harvard Univ, Sch Med, Boston Childrens Hosp, Boston, MA USA
[3] Columbia Univ, Med Ctr, Dept Pediat, New York, NY 10027 USA
关键词
MAGNETIC-RESONANCE INVESTIGATIONS; ACTINOMYCIN-D; EWINGS-SARCOMA; AQUEOUS-SOLUTION; ONCOGENIC TRANSCRIPTION; COMPLEX-FORMATION; GROWTH-FACTOR; GENE; FUSION; TUMOR;
D O I
10.1371/journal.pone.0069714
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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.
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页数:7
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