Functional and regulatory analysis of the Arabidopsis thaliana CAX2 cation transporter.
The vacuolar sequestration of metals is an important metal tolerance mechanism in plants. The Arabidopsis thaliana vacuolar transporters CAX1 and CAX2 were originally identified in a Saccharomyces cerevisiae suppression screen as Ca2+/H+ antiporters. CAX2 has a low affinity for Ca2+ but can transport other metals including Mn2+ and Cd2+. Here we demonstrate that unlike cax1 mutants, CAX2 insertional mutants caused no discernable morphological phenotypes or alterations in Ca2+/H+ antiport activity. However, cax2 lines exhibited a reduction in vacuolar Mn2+/H+ antiport and, like cax1 mutants, reduced V-type H+ -ATPase (V-ATPase) activity. Analysis of a CAX2 promoter beta-glucoronidase (GUS) reporter gene fusion confirmed that CAX2 was expressed throughout the plant and strongly expressed in flower tissue, vascular tissue and in the apical meristem of young plants. Heterologous expression in yeast identified an N-terminal regulatory region in CAX2, suggesting that Arabidopsis contains multiple cation/H+ antiporters with shared regulatory features. Furthermore, despite significant variations in morphological and biochemical phenotypes, cax1 and cax2 lines both significantly alter V-ATPase activity, hinting at coordinate regulation among transporters driven by H+ gradients and the V-ATPase.
Pittman
Jon K
JK
United States Department of Agriculture/Agricultural Research Service Children's Nutrition Research Center, Baylor College of Medicine, 1100 Bates Street, Houston, TX 77030, USA. jon.pittman@manchester.ac.uk
Shigaki
Toshiro
T
Marshall
Joy L
JL
Morris
Jay L
JL
Cheng
Ning-Hui
NH
Hirschi
Kendal D
KD
D000595:Amino Acid Sequence; D017920:Antiporters / Q000235:genetics* / Q000502:physiology; D017360:Arabidopsis / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000502:physiology; D001692:Biological Transport; D002135:Calcium-Binding Proteins / Q000235:genetics* / Q000502:physiology; D027682:Cation Transport Proteins / Q000235:genetics* / Q000502:physiology; D017353:Gene Deletion; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D005838:Genotype; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D008345:Manganese / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D030821:Plants, Genetically Modified; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D025262:Vacuolar Proton-Translocating ATPases / Q000378:metabolism; D014617:Vacuoles / Q000378:metabolism
D017920:Antiporters; D029681:Arabidopsis Proteins; D002135:Calcium-Binding Proteins; D027682:Cation Transport Proteins; D011993:Recombinant Fusion Proteins; C027614:calcium-hydrogen antiporters; D008345:Manganese; D005966:Glucuronidase; D025262:Vacuolar Proton-Translocating ATPases