All cells are critically dependent on homeostatic mechanisms that regulate the level of cations. Disruptions of ion homeostasis are related to various pathologies. K+, H+ and Na+ are among the most prevalent ions which are essential in cell bioenergetics. Their appropriate concentrations are required for the maintenance of cell volume, cell cycle, internal pH and membrane potential. Various cation transport systems participate in ensuring the optimal intracellular level of alkali-metal cations and protons (pH). The proper function of any transporter is controlled by its expression, biogenesis and degradation, as well as post-translation regulation of its activity. Our recent results concerning cation homeostasis regulation in eukaryotic cells at the level of transporters’ biogenesis and protein-protein interactions will be presented.