title: Genetic evidence for the essential role of PfNT1 in the transport and utilization of xanthine, guanine, guanosine and adenine by Plasmodium falciparum.
abstract: 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 pfnt1Delta 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 pfnt1Delta parasites to transport and utilize adenosine, inosine and hypoxanthine as purine sources. The ability of wild-type and pfnt1Delta 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, pfnt1Delta 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 pfnt1Delta 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.
authors.affiliation: Department of Genetics and Developmental Biology, University of Connecticut Health Center, Farmington, CT 06030, USA.
authors.forename: Kamal
authors.initials: K
authors.lastname: El Bissati
authors.forename: Megan J
authors.initials: MJ
authors.lastname: Downie
authors.forename: Seong-Kyoun
authors.initials: SK
authors.lastname: Kim
authors.forename: Michael
authors.initials: M
authors.lastname: Horowitz
authors.forename: Nicola
authors.initials: N
authors.lastname: Carter
authors.forename: Buddy
authors.initials: B
authors.lastname: Ullman
authors.forename: Choukri
authors.initials: C
authors.lastname: Ben Mamoun
mesh_terms: D000225:Adenine / Q000378:metabolism; D000818:Animals; D001692:Biological Transport; D004912:Erythrocytes / Q000469:parasitology; D017353:Gene Deletion; D006147:Guanine / Q000378:metabolism; D006151:Guanosine / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000235:genetics* / Q000378:metabolism*; D010963:Plasmodium falciparum / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D015800:Protozoan Proteins / Q000235:genetics* / Q000378:metabolism*; D011687:Purines / Q000737:chemistry / Q000378:metabolism*; D019820:Xanthine / Q000378:metabolism
chemical_list: D026901:Membrane Transport Proteins; C407347:NT1 protein, Plasmodium falciparum; D015800:Protozoan Proteins; D011687:Purines; D006151:Guanosine; D019820:Xanthine; D006147:Guanine; D000225:Adenine