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Mol. Cell. Biol., 01 1995, 179-185, Vol 15, No. 1
Copyright © 1995, American Society for Microbiology

Thymidylate synthase binds to c-myc RNA in human colon cancer cells and in vitro

E Chu, T Takechi, KL Jones, DM Voeller, SM Copur, GF Maley, F Maley, S Segal and CJ Allegra
NCI-Navy Medical Oncology Branch, Bethesda, Maryland 20892.

Using an immunoprecipitation-reverse transcription-PCR technique, we characterized a thymidylate synthase (TS) ribonucleoprotein complex in cultured human colon cancer cells that consists of TS protein and the mRNA of the nuclear oncogene c-myc. TS protein is complexed in intact cells with the C-terminal coding region of c-myc mRNA that includes nucleotide positions 1625 to 1790. RNA electrophoretic gel mobility shift assays confirm a specific interaction between TS protein and c- myc mRNA and provide additional evidence that the C-terminal coding region represents an important cis-acting regulatory element. Further evidence demonstrates that the in vitro translational efficiency of c- myc mRNA is inhibited as a result of its direct interaction with TS protein. In addition, the presence of exogenous c-myc mRNA specifically relieves the inhibitory effects of TS protein on TS mRNA translation.


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