An organelle K⁺ channel is required for osmoregulation in Chlamydomonas reinhardtii

Fresh water protozoa and algae face hypotonic challenges in their living environment. Many of them employ a contractile vacuole system to uptake excessive water from the cytoplasm and expel it to the environment to achieve cellular homeostasis. K+, a major osmolyte in contractile vacuole, is predicted to create higher osmolarity for water influx. Molecular mechanisms for K+ permeation through the plasma membrane have been well studied. However, how K+ permeates organelles such as the contractile vacuole is not clear. Here, we show that the six-transmembrane K+ channel KCN11 in Chlamydomonas is exclusively localized to contractile vacuole. Ectopic expression of KCN11 in HEK293T cells results in voltage-gated K+ channel activity. Disruption of the gene or mutation of key residues for K+ permeability of the channel leads to dysfunction of cell osmoregulation in very hypotonic conditions. The contractile cycle is inhibited in the mutant cells with a slower rate of contractile vacuole swelling, leading to cell death. These data demonstrate a new role for six-transmembrane K+ channels in contractile vacuole functioning and provide further insights into osmoregulation mediated by the contractile vacuole.