KCNE1-KCNQ1 osmoregulation by interaction of phosphatidylinositol-4,5-bisphosphate with Mg2+ and polyamines.

KCNQ1 osmosensitivity is of physiological and pathophysiological relevance in epithelial and cardiac cells, but the mechanism involved remains elusive. In COS-7 cells expressing the KCNE1-KCNQ1 fusion protein, extracellular hypoosmolarity and hyperosmolarity modify the channel biophysical parameters. These changes are consistent with hypoosmolarity increasing the level of membrane phosphatidylinositol-4,5-bisphosphate (PIP(2)), which in turn upregulates KCNE1-KCNQ1 channels. We showed that increasing PIP(2) levels with a water-soluble PIP(2) analogue prevented channel upregulation in hypoosmotic condition, suggesting a variation of the channel-PIP(2) interaction during channel osmoregulation. Furthermore, we showed that polyamines and Mg(2+), already known to tonically inhibit KCNQ channels by screening PIP(2) negative charges, are involved in the osmoregulatory process. Indeed, intracellular Mg(2+) removal and polyamines chelation inhibited the channel osmoregulation. Thus, the dilution of those cations during cell swelling might decrease channel inhibition and explain the channel upregulation by hypoosmolarity. To support this idea, we quantified the role of Mg(2+) in the osmodependent channel activity. Direct measurement of intracellular [Mg(2+)] variations during osmotic changes and characterization of the channel Mg(2+) sensitivity showed that Mg(2+) participates significantly to the osmoregulation. Using intracellular solutions that mimic the variation of Mg(2+) and polyamines, we were able to recapitulate the current amplitude variations in response to extracellular osmolarity changes. Altogether, these results support the idea of a modulation of the channel-PIP(2) interactions by Mg(2+) and polyamines during cell volume changes. It is likely that this mechanism applies to other channels that are sensitive to both osmolarity and PIP(2).