COMPARISON OF ADENYLATE-CYCLASE AND CAMP-DEPENDENT PROTEIN-KINASE IN GAMETOCYTOGENIC AND NONGAMETOCYTOGENIC CLONES OF PLASMODIUM-FALCIPARUM

Adenylate cyclase and cAMP-dependent protein kinase activities in gametocytogenic (LES) and nongametocytogenic (T9/96) clones of Plasmodium falciparum were compared to explore the role of cAMP in sexual differentiation of the parasite. Basal adenylate cyclase levels were equivalent in the 2 clones. However, cAMP-dependent histone II-A kinase activity was significantly higher in LE5 than in T9/96 over a range of cAMP concentrations. This difference was due to a decreased V(max) for the enzyme in the nongametocytogenic clone and not to an increased K(a) for cAMP. Examination of parasite cAMP-binding proteins, likely to be kinase regulatory subunits, by both photoaffinity labeling with [P-32]8-N3-cAMP and affinity chromatography of metabolically [S-35]methionine-labeled cytosol of cAMP-agarose revealed a 53-kDa cAMP binding protein in both clones and a 49-kDa cAMP-binding protein in T9/96 that was absent in LE5. Our results suggest that T9/96 has lost the ability to undergo gametocytogenesis due to a substantial decrease in cAMP-dependent protein kinase activity rendering the parasite unable to respond to increased intracellular cAMP levels. Moreover, the reduction in cAMP-dependent protein kinase activity may be due to the presence of an alternative regulatory subunit of the kinase.