The biochemical basis for resistance to adenine arabinoside in a mutant of Toxoplasma gondii.
We have previously reported the isolation and preliminary characterization of a mutant of Toxoplasma gondii that was resistant to adenine arabinoside. Fiftyfold higher concentrations of adenine arabinoside were required to inhibit the growth of the resistant parasite in human fibroblast cultures. To determine the enzymic basis for resistance, we measured the kinases and deaminases that act on adenosine or deoxyadenosine. All of these enzymic activities were found in uninfected human fibroblast cells. The mutant and wild type parasite proved to have similar activities of adenosine deaminase, deoxyadenosine deaminase, and deoxyadenosine kinase. However, the adenine arabinoside resistant mutant had less than 0.1% of the adenosine kinase activity observed in the wild type T. gondii. The mutant parasite is presumably resistant because without adenosine kinase to phosphorylate adenine arabinoside it cannot carry out the first step in the conversion of the analogue to adenine arabinoside triphosphate, the active form. A mutant of 3T6 (mouse) cells previously selected for a loss of adenosine kinase also proved to be resistant to adenine arabinoside.
D000243:Adenosine Deaminase / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D003839:Deoxyadenosines / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004351:Drug Resistance; D006801:Humans; D051379:Mice; D009154:Mutation; D010770:Phosphotransferases / Q000378:metabolism; D014122:Toxoplasma / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D014740:Vidarabine / Q000494:pharmacology*
D003839:Deoxyadenosines; D010770:Phosphotransferases; D000243:Adenosine Deaminase; D014740:Vidarabine