A Novel Mechanism of Ion Homeostasis and Salt Tolerance in Yeast: the Hal4 and Hal5 Protein Kinases Modulate the Trk1-Trk2 Potassium Transporter

Abstract

The regulation of intracellular ion concentrations is a fundamental property of living cells. Although many ion transporters have been identified, the systems that modulate their activity remain largely unknown. We have characterized two partially redundant genes from Saccharomyces cerevisiae, HAL4/SAT4 and HAL5, that encode homologous protein kinases implicated in the regulation of cation uptake. Overexpression of these genes increases the tolerance of yeast cells to sodium and lithium, whereas gene disruptions result in greater cation sensitivity. These phenotypic effects of the mutations correlate with changes in cation uptake and are dependent on a functional Trk1-Trk2 potassium transport system. In addition, hal4 hal5 and trk1 trk2 mutants exhibit similar phenotypes: (i) they are deficient in potassium uptake; (ii) their growth is sensitive to a variety of toxic cations, including lithium, sodium, calcium, tetramethylammonium, hygromycin B, and low pH; and (iii) they exhibit increased uptake of methylammonium, an indicator of membrane potential. These results suggest that the Hal4 and Hal5 protein kinases activate the Trk1-Trk2 potassium transporter, increasing the influx of potassium and decreasing the membrane potential. The resulting loss in electrical driving force reduces the uptake of toxic cations and improves salt tolerance. Our data support a role for regulation of membrane potential in adaptation to salt stress that is mediated by the Hal4 and Hal5 kinases.

ACKNOWLEDGMENTS

This work was supported by grants from the Spanish CICYT (Madrid, BIO96-1196) and European Union (Brussels, BIO4-CT96-0775) to R.S. and by NIH grant GM40266 to G.R.F. J.M.M. is a fellow of the Conselleria de Educacio i Ciencia (Valencia), and G.R. was a fellow of the Spanish Ministerio de Educacion y Ciencia (Madrid). S.J.K. was supported in part by a fellowship from the Helen Hay Whitney Foundation.

We thank A. Rodriguez-Navarro (Madrid) for the WΔ3 strain and Avelino Corma (Instituto de Tecnologia Quimica, Valencia, Spain) for making available his atomic absorption spectrophotometer.

J.M.M. and M.P.L. contributed equally to this work.