The TRK1 potassium transporter is the critical effector for killing of Candida albicans by the cationic protein, histatin 5

The principal feature of killing of Candida albicans and other pathogenic fungi by the calonic protein Histatin 5 (Hst 5) is loss of cytoplasmic small molecules and ions, including ATP and K⁺, which can be blocked by the anion channel inhibitor 4,4′-diisothiocyanatostilbene-2,2′- disulfonic acid. We constructed C. albicans strains expressing one, two, or three copies of the TRK1 gene in order to investigate possible roles of Trk1p (the organism's principal K⁺ transporter) in the actions of Hst 5. All measured parameters (Hst 5 killing, Hst 5-stimulated ATP efflux, normal Trk1p-mediated K⁺ (⁸⁶Rb⁺) influx, and Trk1p-mediated chloride conductance) were similarly reduced (5-7-fold) by removal of a single copy of the TRK1 gene from this diploid organism and were fully restored by complementation of the missing allele. A TRK1 overexpression strain of C. albicans, constructed by integrating an additional TRK1 gene into wild-type cells, demonstrated cytoplasmic sequestration of Trk1 protein, along with somewhat diminished toxicity of Hst 5. These results could be produced either by depletion of intracellular free Hst 5 due to sequestered binding, or to cooperativity in Hst 5-protein interactions at the plasma membrane. Furthermore, Trk1p-mediated chloride conductance was blocked by 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid in all of the tested strains, strongly suggesting that the TRK1 protein provides the essential pathway for ATP loss and is the critical effector for Hst 5 toxicity in C. albicans.