The molecular events which confer cellular resistance to cytotoxic drugs such as cytosine arabinoside (ara-C) are poorly understood. Nevertheless, in a proportion of patients with acute leukemia, such events will be responsible for the failure of therapy. Mutations which cause ara-C resistance in a chinese hamster ovary (CHO) cell model have been identified as regulatory base substitutions, occurring in specific sites of the gene coding for an enzyme critical in pyrimidine metabolism, CTP synthetase (CTPs). These cells have elevated dCTP pools, a feature common to biochemical studies of other ara-C resistant leukemic cells. A 94% homology exists between the hamster and human ctps genes. In this study, similar mutations were sought in samples taken from 36 patients, with recurrent or resistant acute leukemia. No mutations were identified in the regions indicated by the CHO model using techniques capable of detecting mutations only if present in more than 10% of the cells studied. Thus, mutations in these sites within the human ctps gene do not appear to be a major mechanism of resistance to ara-C in acute leukemia. Further studies should be directed towards developing more sensitive methods of detection, and these then applied both to CTPs and to other enzymes involved in pyrimidine metabolism.