title: Adenine phosphoribosyltransferase-deficient Leishmania donovani.
abstract: Mutant promastigotes of Leishmania donovani deficient in adenine phosphoribosyltransferase (APRTase) have been isolated in medium containing 4-aminopyrazolopyrimidine. The generation of APRTase-deficient mutants occurred in two discrete steps. In the first step, clones were isolated with 50% of wildtype levels of APRTase activity. These cells were reselected and colonies totally deficient in APRTase were isolated. Partially and totally APRTase-deficient cells exhibited intermediate and complete resistance to cytotoxic adenine analogs, respectively. Nevertheless, wildtype and mutant cells could salvage adenine and utilize adenine as a purine source equally efficiently, suggesting that the adenine deaminase-HGPRTase pathway plays an important role in promastigote adenine metabolism. Kinetic and thermal inactivation studies of purified APRTase and isoelectric focusing of crude extracts from wildtype and partially APRTase-deficient cells suggested that the latter cells possessed wildtype APRTase activity at half the amount found in wildtype parental cells. These data suggest that Leishmania donovani possess two copies of the APRTase structural gene and that these organisms might be diploid for the APRTase locus.
mesh_terms: D000225:Adenine / Q000378:metabolism*; D000228:Adenine Phosphoribosyltransferase / Q000172:deficiency* / Q000235:genetics; D000619:Aminohydrolases / Q000378:metabolism; D000818:Animals; D007041:Hypoxanthine Phosphoribosyltransferase / Q000378:metabolism; D007893:Leishmania donovani / Q000201:enzymology* / Q000235:genetics; D009154:Mutation; D010430:Pentosyltransferases / Q000172:deficiency*
chemical_list: D010430:Pentosyltransferases; D000228:Adenine Phosphoribosyltransferase; D007041:Hypoxanthine Phosphoribosyltransferase; D000619:Aminohydrolases; C018774:adenine deaminase; D000225:Adenine