Genetic evidence for the essential role of PfNT1 in the transport and utilization of xanthine, guanine, guanosine and adenine by Plasmodium falciparum

• Published in: Molecular and biochemical parasitology 2008-10, Vol.161 (2), p.130-139
• Main Authors: Bissati, Kamal El, Downie, Megan J., Kim, Seong-Kyoun, Horowitz, Michael, Carter, Nicola, Ullman, Buddy, Mamoun, Choukri Ben
• Format: Article
• Language: English
• Subjects: Adenine - metabolism; Animals; Biological Transport; Erythrocytes - parasitology; Gene Deletion; Guanine - metabolism; Guanosine - metabolism; Malaria; Membrane Transport Proteins - genetics; Membrane Transport Proteins - metabolism; PfNT1; Plasmodium falciparum; Plasmodium falciparum - genetics; Plasmodium falciparum - growth & development; Plasmodium falciparum - metabolism; Protozoan Proteins - genetics; Protozoan Proteins - metabolism; Purine; Purines - chemistry; Purines - metabolism; Transport; Xanthine - metabolism
• ISSN: 0166-6851; 1872-9428
• DOI: 10.1016/j.molbiopara.2008.06.012

The malaria parasite, Plasmodium falciparum, is unable to synthesize the purine ring de novo and is therefore wholly dependent upon purine salvage from the host for survival. Previous studies have indicated that a P. falciparum strain in which the purine transporter PfNT1 had been disrupted was unable to grow on physiological concentrations of adenosine, inosine and hypoxanthine. We have now used an episomally complemented pfnt1Δ knockout parasite strain to confirm genetically the functional role of PfNT1 in P. falciparum purine uptake and utilization. Episomal complementation by PfNT1 restored the ability of pfnt1Δ parasites to transport and utilize adenosine, inosine and hypoxanthine as purine sources. The ability of wild-type and pfnt1Δ knockout parasites to transport and utilize the other physiologically relevant purines adenine, guanine, guanosine and xanthine was also examined. Unlike wild-type and complemented P. falciparum parasites, pfnt1Δ parasites could not proliferate on guanine, guanosine or xanthine as purine sources, and no significant transport of these substrates could be detected in isolated parasites. Interestingly, whereas isolated pfnt1Δ parasites were still capable of adenine transport, these parasites grew only when adenine was provided at high, non-physiological concentrations. Taken together these results demonstrate that, in addition to hypoxanthine, inosine and adenosine, PfNT1 is essential for the transport and utilization of xanthine, guanine and guanosine.