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First published online February 28, 2002; 10.1104/pp.010857

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Plant Physiol, April 2002, Vol. 128, pp. 1245-1254

Characterization of CAX4, an Arabidopsis H+/Cation Antiporter1

Ning-hui Cheng, Jon K. Pittman, Toshiro Shigaki, and Kendal D. Hirschi*

Baylor College of Medicine, Plant Physiology Group, United States Department of Agriculture/Agricultural Research Service Children's Nutrition Research Center, 1100 Bates Street, Houston, Texas 77030 (N.-h.C., J.K.P., T.S., K.D.H.); and Vegetable and Fruit Improvement Center, Texas A&M University, College Station, Texas 77845 (K.D.H.)

Ion compartmentalization is essential for plant growth and development. The Arabidopsis open reading frames for CAX1, CAX2, and CAX3 (cation exchangers 1, 2, and 3) were previously identified as transporters that may modulate ion fluxes across the vacuolar membrane. To understand the diversity and role of H+/cation transporters in controlling plant ion levels, another homolog of the CAX genes, CAX4, was cloned from an Arabidopsis cDNA library. CAX4 is 53% identical to CAX1 at the amino acid level, 42% identical to CAX2, and 54% identical to CAX3. CAX4 transcripts appeared to be expressed at low levels in all tissues and levels of CAX4 RNA increased after Mn2+, Na+, and Ni2+ treatment. An N-terminal CAX4-hemagglutinin fusion appeared to localize to both yeast and plant vacuolar membranes. When expressed in yeast, CAX4, like CAX3, failed to suppress the Ca2+ sensitivity of yeast strains deficient in vacuolar Ca2+ transport. Several modifications to CAX4 allowed the protein to transport Ca2+. Addition of amino acids to the N terminus of CAX4 and CAX3 caused both transporters to suppress the sensitivity of yeast strains deficient in vacuolar Ca2+ transport. These findings suggest that CAX transporters may modulate their ion transport properties through alterations at the N terminus.

1 This work was supported by the U.S. Department of Agriculture/Agricultural Research Service (cooperative agreement no. 58-6250-6001), by Phillip Morris USA, and by the National Institutes of Health (grant nos. CHRC 5 P30 and 1R01 GM57427).

* Corresponding author; e-mail kendalh{at}bcm.tmc.edu; fax 713-798-7078.

© 2002 American Society of Plant Physiologists


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