Facilitation by pyrimidine deoxyribonucleosides and hypoxanthine of mutagenic and cytotoxic effects of monofunctional alkylating agents in Chinese hamster cells

Abstract

Hypoxanthine (Hx), thymidine (TdR) and deoxycytidine (CdR), at concentrations of 10 −5 M increased the yield of 8-azaguanine-resistant (AzG r) mutants induced by N-methyl- N′-nitro- N-nitrosoguanidine (MNNG) in cultured Chinese hamster V79 cells. The cytotoxicity of MNNG was increased 2-fold in the presence of Hx, and more than 10-fold in the presence of TdR. This cytotoxic effect of TdR was abolished by equal concentrations of CdR, which by itself did not increase the cytotoxicity of MNNG. However, the yield of MNNG-induced AzG r colonies was increased 2–10-fold in the presence of both CdR and TdR. The AzG r colonies displayed phenotypes characteristic of hypoxanthine: guaninephosphoribosyltransferase-deficient (HGPRT −) mutants, or indicative of mutant HGPRT with altered substrate affinities. The nucleotides did not affect the growth or expression time of the HGPRT − mutants; the same extent of alkali-labile DNA occurred in cells treated with alkylating agents in the presence and absence of TdR and CdR; and the increase in mutation frequency in the presence of these nucleosides occurred not only with MNNG, but also with ethylating agents. Nucleosides treated with MNNG were not mutagenic, and treatment of the cells with TdR and CdR only prior to treatment with MNNG or only during selection with AzG did not increase the induced mutation frequency. Therefore, the interpretation is proposed that CdR, TdR and Hx produce nucleotide-pool imbalances that increase lethal and mutagenic errors of replication of alkylated DNA.