title: The Trypanosoma cruzi Diamine Transporter Is Essential for Robust Infection of Mammalian Cells.
abstract: Trypanosoma cruzi is incapable of synthesizing putrescine or cadaverine de novo, and, therefore, salvage of polyamines from the host milieu is an obligatory nutritional function for the parasite. A high-affinity diamine transporter (TcPOT1) from T. cruzi has been identified previously that recognizes both putrescine and cadaverine as ligands. In order to assess the functional role of TcPOT1 in intact parasites, a Δtcpot1 null mutant was constructed by targeted gene replacement and characterized. The Δtcpot1 mutant lacked high-affinity putrescine-cadaverine transport capability but retained the capacity to transport diamines via a non-saturable, low-affinity mechanism. Transport of spermidine and arginine was not impacted by the Δtcpot1 lesion. The Δtcpot1 cell line exhibited a significant but not total defect in its ability to subsist in Vero cells, although initial infection rates were not affected by the lesion. These findings reveal that TcPOT1 is the sole high-affinity diamine permease in T. cruzi, that genetic obliteration of TcPOT1 impairs the ability of the parasite to maintain a robust infection in mammalian cells, and that a secondary low-affinity uptake mechanism for this key parasite nutrient is operative but insufficient for optimal infection.
authors.affiliation: Department of Biochemistry and Molecular Biology, Oregon Health & Science University, Portland, Oregon, United States of America.
authors.forename: Marie-Pierre
authors.initials: MP
authors.lastname: Hasne
authors.affiliation: Department of Biochemistry and Molecular Biology, Oregon Health & Science University, Portland, Oregon, United States of America.
authors.forename: Radika
authors.initials: R
authors.lastname: Soysa
authors.affiliation: Department of Biochemistry and Molecular Biology, Oregon Health & Science University, Portland, Oregon, United States of America.
authors.forename: Buddy
authors.initials: B
authors.lastname: Ullman
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003959:Diamines / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000378:metabolism*; D014349:Trypanosoma cruzi / Q000378:metabolism* / Q000472:pathogenicity; D014774:Virulence
chemical_list: D003959:Diamines; D026901:Membrane Transport Proteins