Distinct N-terminal regulatory domains of Ca(2+)/H(+) antiporters

• Published in: Plant physiology (Bethesda) Vol. 130; no. 2; p. 1054
• Main Authors: Pittman, Jon K, Sreevidya, Coimbatore S, Shigaki, Toshiro, Ueoka-Nakanishi, Hanayo, Hirschi, Kendal D
• Format: Journal Article
• Language: English
• Published: United States 01.10.2002
• Subjects: Amino Acid Sequence; Antiporters - genetics; Antiporters - metabolism; Arabidopsis - genetics; Arabidopsis - metabolism; Biological Transport - drug effects; Calcium - metabolism; Calcium - pharmacology; Calcium-Binding Proteins - genetics; Calcium-Binding Proteins - metabolism; Cation Transport Proteins; Cloning, Molecular; Fabaceae - genetics; Fabaceae - metabolism; Gene Expression Regulation, Plant; Histidine - pharmacology; Molecular Sequence Data; Mutagenesis, Site-Directed; Mutation; Nitrogen - metabolism; Peptide Fragments - genetics; Peptide Fragments - metabolism; Saccharomyces cerevisiae - genetics; Sequence Homology, Amino Acid; Vacuoles - metabolism
• ISSN: 0032-0889

The regulation of intracellular Ca(2+) levels is achieved in part by high-capacity vacuolar Ca(2+)/H(+) antiporters. An N-terminal regulatory region (NRR) on the Arabidopsis Ca(2+)/H(+) antiporter CAX1 (cation exchanger 1) has been shown previously to regulate Ca(2+) transport by a mechanism of N-terminal auto-inhibition. Here, we examine the regulation of other CAX transporters, both within Arabidopsis and from another plant, mung bean (Vigna radiata), to ascertain if this mechanism is commonly used among Ca(2+)/H(+) antiporters. Biochemical analysis of mung bean VCAX1 expressed in yeast (Saccharomyces cerevisiae) showed that N-terminal truncated VCAX1 had approximately 70% greater antiport activity compared with full-length VCAX1. A synthetic peptide corresponding to the NRR of CAX1, which can strongly inhibit Ca(2+) transport by CAX1, could not dramatically inhibit Ca(2+) transport by truncated VCAX1. The N terminus of Arabidopsis CAX3 was also shown to contain an NRR. Additions of either the CAX3 or VCAX1 regulatory regions to the N terminus of an N-terminal truncated CAX1 failed to inhibit CAX1 activity. When fused to N-terminal truncated CAX1, both the CAX3 and VCAX1 regulatory regions could only auto-inhibit CAX1 after mutagenesis of specific amino acids within this NRR region. These findings demonstrate that N-terminal regulation is present in other plant CAX transporters, and suggest distinct regulatory features among these transporters.