The serum- and glucocorticoid-inducible kinase SGK1 has been shown to regulate a variety of ion channels including the epithelial Ca²⁺ channels TRPV5 and TRPV6. The effect of SGK1 on TRPV5 but not on TRPV6 required the coexpression of NHERF2. The mechanism of TRPV6 regulation remained elusive. SGK1 phosphorylates and thus activates the mammalian phosphatidylinositol-3-phosphate-5-kinase PIKfyve, which in turn stimulates several transport proteins. The present study utilized heterologous expression in Xenopus oocytes to explore whether PIKfyve participates in the regulation of TRPV6. Ca²⁺ entry though this channel activates endogenous Ca²⁺-sensitive Cl^- channels, which are determined by dual electrode voltage clamp. Expression of wild type PIKfyve markedly increased the conductance of oocytes injected with TRPV6 mRNA but not of oocytes injected without TRPV6 mRNA. Coexpression of constitutive active ^S422DSGK1 similarly enhanced TRPV6- dependent currents. In contrast, coexpression of the SGK1 resistant ^S31APIKfyve mutant did not stimulate TRPV6-dependent currents. Coexpression of both, ^S422DSGK1 and PIKfyve, was followed by a current significantly larger than the current induced by either, ^S422DSGK1 or PIKfyve, alone. In conclusion, PIKfyve indeed stimulates the Ca²⁺ channel TRPV6.