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J. Biol. Chem., Vol. 260, Issue 30, 16274-16278, Dec, 1985

Thymidine incorporation in nucleoside transport-deficient lymphoma cells

B Aronow and B Ullman

Nucleoside transport deficiency in mammalian cells is associated with an inability to transport most nucleosides, growth resistance to a spectrum of cytotoxic nucleosides, and a loss of binding sites for 4- nitrobenzylthioinosine (NBMPR), a potent inhibitor of nucleoside transport. The nucleoside transport-deficient S49 T lymphoma cell line, AE1, however, was almost as capable of incorporating thymidine into TTP as the wild type parent provided thymidine was administered at a sufficiently high concentration. Consequently, AE1 cells were just as sensitive as wild type cells to the toxicity of high thymidine concentrations. In contrast, AE1 cells were highly resistant to almost all other cytotoxic nucleosides including the thymidine analogs, 5- bromodeoxyuridine and 5-fluoro-2'-deoxyuridine 5'-monophosphate. Despite having demonstrable ability to accumulate TTP, AE1 cells were unable to grow on hypoxanthine-amethopterin-thymidine (HAT)-containing medium. This was due to their inability to accumulate sufficient TTP from the low concentrations of thymidine present in HAT medium. AE1 cells possessed an incomplete thymidine transport deficiency, the extent of which was concentration dependent. The residual capacity for thymidine transport present in AE1 cells was insensitive to inhibition by 4-nitrobenzylthioinosine and could account both for their inability to grow on HAT medium and their sensitivity to cytotoxic concentrations of thymidine. Another nucleoside transport-deficient cell line, FURD-80- 3-6, was similar to the AE1 cell line in its growth phenotype and NBMPR- binding site deficiency but differed in its decreased growth sensitivity to thymidine. That nucleoside transport deficiencies may vary in their completeness for different nucleosides has significance for the mechanism by which a single transporter can recognize a wide variety of nucleosides.
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