The biochemical characterization of two phosphate transport systems in Phytomonas serpens

Inorganic phosphate (P_i) is an essential nutrient for all organisms because it is required for a variety of biochemical processes, such as signal transduction and the synthesis of phosphate-containing biomolecules. Assays of ³²P_i uptake performed in the absence or in the presence of Na⁺ indicated the existence of a Na⁺-dependent and a Na⁺-independent P_i transporter in Phytomonas serpens. Phylogenetic analysis of two hypothetical protein sequences of Phytomonas (EM1) showed similarities to the high-affinity P_i transporters of Saccharomyces cerevisiae: Pho84, a Na⁺-independent P_i transporter, and Pho89, a Na⁺-dependent P_i transporter. Plasma membrane depolarization by FCCP, an H⁺ ionophore, strongly decreased P_i uptake via both Na⁺-independent and Na⁺-dependent carriers, indicating that a membrane potential is essential for P_i influx. In addition, the furosemide-sensitive Na⁺-pump activity in the cells grown in low P_i conditions was found to be higher than the activity detected in the plasma membrane of cells cultivated at high P_i concentration, suggesting that the up-regulation of the Na⁺-ATPase pump could be related to the increase of P_i uptake by the Pho89p Na⁺:P_i symporter. Here we characterize for the first time two inorganic phosphate transporters powered by Na⁺ and H⁺ gradients and activated by low P_i availability in the phytopathogen P. serpens.