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Plant Physiol, December 2000, Vol. 124, pp. 1814-1827

The ACA4 Gene of Arabidopsis Encodes a Vacuolar Membrane Calcium Pump That Improves Salt Tolerance in Yeast1

Markus Geisler,2 Nathalie Frangne, Eric Gomès, Enrico Martinoia, and Michael G. Palmgren*

Department of Plant Biology, The Royal Veterinary and Agricultural University, Thorvaldsensvej 40, DK-1871 Frederiksberg C, Denmark (M.G., E.G., M.G.P.); and Institut de Botanique, Physiologie Végétale, Université de Neuchâtel, 2007 Neuchatel, Switzerland (N.F., E.M.)

Several lines of evidence suggest that regulation of intracellular Ca2+ levels is crucial for adaptation of plants to environmental stress. We have cloned and characterized Arabidopsis auto-inhibited Ca2+-ATPase, isoform 4 (ACA4), a calmodulin-regulated Ca2+-ATPase. Confocal laser scanning data of a green fluorescent protein-tagged version of ACA4 as well as western-blot analysis of microsomal fractions obtained from two-phase partitioning and Suc density gradient centrifugation suggest that ACA4 is localized to small vacuoles. The N terminus of ACA4 contains an auto-inhibitory domain with a binding site for calmodulin as demonstrated through calmodulin-binding studies and complementation experiments using the calcium transport yeast mutant K616. ACA4 and PMC1, the yeast vacuolar Ca2+-ATPase, conferred protection against osmotic stress such as high NaCl, KCl, and mannitol when expressed in the K616 strain. An N-terminally modified form of ACA4 specifically conferred increased NaCl tolerance, whereas full-length ATPase had less effect.

1 This work was supported in part by the European Union Biotechnology program, the Danish Biotech III program, and the Alexander von Humboldt Foundation (Feodor-Lynen Fellowship to M.G.).

2 Present address: Institut de Botanique, Physiologie Végétale, Université de Neuchâtel, 2007 Neuchâtel, Switzerland.

* Corresponding author; e-mail palmgren{at}biobase.dk; fax 45-3528-3333.

© 2000 American Society of Plant Physiologists


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