Abstract
Reestablishment of the resting state after stimulus-coupled elevations of cytosolic-free Ca2+ requires the rapid removal of Ca2+ from the cytosol of plant cells. Here we describe the isolation of two genes, CAX1 and CAX2, from Arabidopsis thaliana that suppress a mutant of Saccharomyces cerevisiae that has a defect in vacuolar Ca2+ accumulation. Both genes encode polypeptides showing sequence similarities to microbial H+/Ca2+ antiporters. Experiments on vacuolar membrane-enriched vesicles isolated from yeast expressing CAX1 or CAX2 demonstrate that these genes encode high efficiency and low efficiency H+/Ca2+ exchangers, respectively. The properties of the CAX1 gene product indicate that it is the high capacity transporter responsible for maintaining low cytosolic-free Ca2+ concentrations in plant cells by catalyzing pH gradient-energized vacuolar Ca2+ accumulation.
Full text
PDF![8782](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eda5/38751/644e2f5c5f23/pnas01520-0638.png)
![8783](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eda5/38751/0fa1cfd94b79/pnas01520-0639.png)
![8784](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eda5/38751/c13126b05a29/pnas01520-0640.png)
![8785](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eda5/38751/8529cf5d7968/pnas01520-0641.png)
![8786](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eda5/38751/cd5c8cf737ad/pnas01520-0642.png)
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Allen G. J., Muir S. R., Sanders D. Release of Ca2+ from individual plant vacuoles by both InsP3 and cyclic ADP-ribose. Science. 1995 May 5;268(5211):735–737. doi: 10.1126/science.7732384. [DOI] [PubMed] [Google Scholar]
- Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
- Berridge M. J. Cell signalling. A tale of two messengers. Nature. 1993 Sep 30;365(6445):388–389. doi: 10.1038/365388a0. [DOI] [PubMed] [Google Scholar]
- Blackford S., Rea P. A., Sanders D. Voltage sensitivity of H+/Ca2+ antiport in higher plant tonoplast suggests a role in vacuolar calcium accumulation. J Biol Chem. 1990 Jun 15;265(17):9617–9620. [PubMed] [Google Scholar]
- Blumwald E., Poole R. J. Kinetics of Ca/H Antiport in Isolated Tonoplast Vesicles from Storage Tissue of Beta vulgaris L. Plant Physiol. 1986 Mar;80(3):727–731. doi: 10.1104/pp.80.3.727. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boeke J. D., LaCroute F., Fink G. R. A positive selection for mutants lacking orotidine-5'-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance. Mol Gen Genet. 1984;197(2):345–346. doi: 10.1007/BF00330984. [DOI] [PubMed] [Google Scholar]
- Bowler C., Neuhaus G., Yamagata H., Chua N. H. Cyclic GMP and calcium mediate phytochrome phototransduction. Cell. 1994 Apr 8;77(1):73–81. doi: 10.1016/0092-8674(94)90236-4. [DOI] [PubMed] [Google Scholar]
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Cunningham K. W., Fink G. R. Calcineurin inhibits VCX1-dependent H+/Ca2+ exchange and induces Ca2+ ATPases in Saccharomyces cerevisiae. Mol Cell Biol. 1996 May;16(5):2226–2237. doi: 10.1128/mcb.16.5.2226. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cunningham K. W., Fink G. R. Calcineurin-dependent growth control in Saccharomyces cerevisiae mutants lacking PMC1, a homolog of plasma membrane Ca2+ ATPases. J Cell Biol. 1994 Feb;124(3):351–363. doi: 10.1083/jcb.124.3.351. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Galione A., Lee H. C., Busa W. B. Ca(2+)-induced Ca2+ release in sea urchin egg homogenates: modulation by cyclic ADP-ribose. Science. 1991 Sep 6;253(5024):1143–1146. doi: 10.1126/science.1909457. [DOI] [PubMed] [Google Scholar]
- Gehring C. A., Williams D. A., Cody S. H., Parish R. W. Phototropism and geotropism in maize coleoptiles are spatially correlated with increases in cytosolic free calcium. Nature. 1990 Jun 7;345:528–530. doi: 10.1038/345528a0. [DOI] [PubMed] [Google Scholar]
- Gilroy S., Bethke P. C., Jones R. L. Calcium homeostasis in plants. J Cell Sci. 1993 Oct;106(Pt 2):453–461. doi: 10.1242/jcs.106.2.453. [DOI] [PubMed] [Google Scholar]
- Higgins D. G., Sharp P. M. CLUSTAL: a package for performing multiple sequence alignment on a microcomputer. Gene. 1988 Dec 15;73(1):237–244. doi: 10.1016/0378-1119(88)90330-7. [DOI] [PubMed] [Google Scholar]
- Ivey D. M., Guffanti A. A., Zemsky J., Pinner E., Karpel R., Padan E., Schuldiner S., Krulwich T. A. Cloning and characterization of a putative Ca2+/H+ antiporter gene from Escherichia coli upon functional complementation of Na+/H+ antiporter-deficient strains by the overexpressed gene. J Biol Chem. 1993 May 25;268(15):11296–11303. [PubMed] [Google Scholar]
- Kim E. J., Zhen R. G., Rea P. A. Site-directed mutagenesis of vacuolar H(+)-pyrophosphatase. Necessity of Cys634 for inhibition by maleimides but not catalysis. J Biol Chem. 1995 Feb 10;270(6):2630–2635. doi: 10.1074/jbc.270.6.2630. [DOI] [PubMed] [Google Scholar]
- Knight M. R., Campbell A. K., Smith S. M., Trewavas A. J. Transgenic plant aequorin reports the effects of touch and cold-shock and elicitors on cytoplasmic calcium. Nature. 1991 Aug 8;352(6335):524–526. doi: 10.1038/352524a0. [DOI] [PubMed] [Google Scholar]
- Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
- Minet M., Dufour M. E., Lacroute F. Complementation of Saccharomyces cerevisiae auxotrophic mutants by Arabidopsis thaliana cDNAs. Plant J. 1992 May;2(3):417–422. doi: 10.1111/j.1365-313x.1992.00417.x. [DOI] [PubMed] [Google Scholar]
- Niyogi K. K., Fink G. R. Two anthranilate synthase genes in Arabidopsis: defense-related regulation of the tryptophan pathway. Plant Cell. 1992 Jun;4(6):721–733. doi: 10.1105/tpc.4.6.721. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schumaker K. S., Sze H. A Ca/H Antiport System Driven by the Proton Electrochemical Gradient of a Tonoplast H-ATPase from Oat Roots. Plant Physiol. 1985 Dec;79(4):1111–1117. doi: 10.1104/pp.79.4.1111. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schumaker K. S., Sze H. Solubilization and reconstitution of the oat root vacuolar h/ca exchanger. Plant Physiol. 1990 Feb;92(2):340–345. doi: 10.1104/pp.92.2.340. [DOI] [PMC free article] [PubMed] [Google Scholar]