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[Cancer Research 63, 4268-4274, July 15, 2003]
© 2003 American Association for Cancer Research

Tumor Biology

Functional Analysis of the EWS/ETS Target Gene Uridine Phosphorylase1

Benjamin Deneen, Habib Hamidi and Christopher T. Denny2

Molecular Biology Institute [B. D., C. T. D.], Department of Pediatrics, Gwynne Hazen Cherry Memorial Laboratories [H. H., C. T. D.], and Jonsson Comprehensive Cancer Center [C. T. D.], University of California Los Angeles, Los Angeles, California 90024

The EWS/ETS fusion proteins associated with Ewings family tumors (EFTs) are thought to promote oncogenesis by acting as aberrant transcription factors. Uridine phosphorylase is a gene that is up-regulated by structurally distinct EWS/ETS fusions. Ectopic expression of uridine phosphorylase was able to support anchorage-independent cell growth, indicating that it plays an active role in the oncogenic process. Transcriptional up-regulation of uridine phosphorylase is shown to be mediated in a DNA binding-dependent manner, and reporter gene assays demonstrated that EWS/FLI1 and RAS mediate activation through a single activator protein 1/ETS site located in the uridine phosphorylase promoter. Chromatin immunoprecipitation assays reveal that EWS/FLI1 directly associates with the uridine phosphorylase promoter in vivo. Up-regulation of uridine phosphorylase by EWS/FLI1 sensitizes cells to growth inhibition by the pyrimidine analogue, 5'-deoxy-5'fluorouridine, both in tissue culture and in vivo model systems.




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