Arginine Kinase Overexpression Improves Trypanosoma cruzi Survival Capability

ALONSO, GUILLERMO; IVALDI, SOLEDAD; SILBER, ARIEL; BOUVIER, LEON; ALVES, MARIA JULIA MANSO; TORRES, HECTOR; FLAWIA, MIRTHA; PEREIRA, CA

Caxambu

Congreso; XIX, Annual Meeting of The Brazilian Society of Protozoology; 2003

Arginine kinase catalyzes the reversible transphosphorylation between phosphoarginine and ADP. Phosphoarginine is a metabolite that is involved in temporal and spatial ATP buffering as well as inorganic phosphate regulation. Recently, we demonstrated that arginine kinase is present in the parasites Trypanosoma cruzi and Trypanosoma brucei. In turn, this enzyme is absent in their mammalian hosts. In this work we establish a relationship among the homologous overexpression of the Trypanosoma cruzi arginine kinase and the ability of the transfectant parasite cells to grow and resist stress conditions. Using the novel expression vector pTREX it was obtained more than one hundredfold overexpression of the T. cruzi arginine kinase activity. The stable-transfected parasites showed an increased cell density since day 10 of culture, which resulted about 2.5-fold higher than the control group on day 28. These results suggest an improved capability of the transfected parasites to survive in a nutrient-depleted medium. Additional stress conditions were tested by incubating the parasites in different media. Arginine kinase transfected parasites revealed about 54% of increase in the cell number growing in Triatomine Artificial Urine medium, 91% in alkaline PBS and 79% in conditioned medium. We propose that arginine kinase and phosphoarginine are involved in the regulation of the Trypanosoma cruzi growth and its adaptation to environmental changes.and Trypanosoma brucei. In turn, this enzyme is absent in their mammalian hosts. In this work we establish a relationship among the homologous overexpression of the Trypanosoma cruzi arginine kinase and the ability of the transfectant parasite cells to grow and resist stress conditions. Using the novel expression vector pTREX it was obtained more than one hundredfold overexpression of the T. cruzi arginine kinase activity. The stable-transfected parasites showed an increased cell density since day 10 of culture, which resulted about 2.5-fold higher than the control group on day 28. These results suggest an improved capability of the transfected parasites to survive in a nutrient-depleted medium. Additional stress conditions were tested by incubating the parasites in different media. Arginine kinase transfected parasites revealed about 54% of increase in the cell number growing in Triatomine Artificial Urine medium, 91% in alkaline PBS and 79% in conditioned medium. We propose that arginine kinase and phosphoarginine are involved in the regulation of the Trypanosoma cruzi growth and its adaptation to environmental changes.Trypanosoma cruzi arginine kinase and the ability of the transfectant parasite cells to grow and resist stress conditions. Using the novel expression vector pTREX it was obtained more than one hundredfold overexpression of the T. cruzi arginine kinase activity. The stable-transfected parasites showed an increased cell density since day 10 of culture, which resulted about 2.5-fold higher than the control group on day 28. These results suggest an improved capability of the transfected parasites to survive in a nutrient-depleted medium. Additional stress conditions were tested by incubating the parasites in different media. Arginine kinase transfected parasites revealed about 54% of increase in the cell number growing in Triatomine Artificial Urine medium, 91% in alkaline PBS and 79% in conditioned medium. We propose that arginine kinase and phosphoarginine are involved in the regulation of the Trypanosoma cruzi growth and its adaptation to environmental changes.T. cruzi arginine kinase activity. The stable-transfected parasites showed an increased cell density since day 10 of culture, which resulted about 2.5-fold higher than the control group on day 28. These results suggest an improved capability of the transfected parasites to survive in a nutrient-depleted medium. Additional stress conditions were tested by incubating the parasites in different media. Arginine kinase transfected parasites revealed about 54% of increase in the cell number growing in Triatomine Artificial Urine medium, 91% in alkaline PBS and 79% in conditioned medium. We propose that arginine kinase and phosphoarginine are involved in the regulation of the Trypanosoma cruzi growth and its adaptation to environmental changes.Trypanosoma cruzi growth and its adaptation to environmental changes.growth and its adaptation to environmental changes.