Ion imbalances are the predominant environmental stress that plants have to overcome. Plants must maintain a precise balance of cytosolic and extracellular ion concentrations despite widely varying soil conditions. Vacuolar ion transporters, including calcium (Ca2+) transporter, are key mediators of cytosolic ion concentrations. Intracellular Ca2+ release from vacuoles may also play a centra role in plant signal transduction events. Consequently, vacuolar ion transporters are components of signal transduction events and are necessary for general ion homeostasis during all aspects of growth and development. We have initiated a molecular and genetic approach to understand the processes of ion homeostasis and Ca2+ signal transduction in and around the plant vacuole. We have isolated two genes, designated CAX1 and CAX2 (CAlcium eXchangers 1 and 2), which represent the first vacuolar Ca2+ transporters isolated from plants. Biochemical measurements demonstrate that CAX1p is a Ca2+/H+ antiporter and CAX2p is a vacuolar heavy metal/H+ transporter with weak Ca2+ transport properties. The focus of this proposal is to determine the function and regulation of the CAX genes in plants. Three strategies to achieve these goals will be pursued: a)analysis of CAX expression, regulation and localization will establish their temporal and spatial expression; b)phenotypic analysis of CAX mutants will elucidate their function(s) in plants; and c)identification and characterization of plant CAX regulatory components. Once these transporters have been further characterized and additional regulators of ion homeostasis have been identified, then manipulating plant ion storage, signal transduction events, and altering the environmental constraints of agriculturally important plants, becomes realistic goals.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM057427-04
Application #
6386869
Study Section
Molecular Cytology Study Section (CTY)
Program Officer
Shapiro, Bert I
Project Start
1998-05-01
Project End
2003-04-30
Budget Start
2001-05-01
Budget End
2002-04-30
Support Year
4
Fiscal Year
2001
Total Cost
$164,394
Indirect Cost
Name
Baylor College of Medicine
Department
Pediatrics
Type
Schools of Medicine
DUNS #
074615394
City
Houston
State
TX
Country
United States
Zip Code
77030
Pittman, Jon K; Shigaki, Toshiro; Marshall, Joy L et al. (2004) Functional and regulatory analysis of the Arabidopsis thaliana CAX2 cation transporter. Plant Mol Biol 56:959-71
Magidin, Monica; Pittman, Jon K; Hirschi, Kendal D et al. (2003) ILR2, a novel gene regulating IAA conjugate sensitivity and metal transport in Arabidopsis thaliana. Plant J 35:523-34
Shigaki, Toshiro; Pittman, Jon K; Hirschi, Kendal D (2003) Manganese specificity determinants in the Arabidopsis metal/H+ antiporter CAX2. J Biol Chem 278:6610-7
Cheng, Ning-Hui; Hirschi, Kendal D (2003) Cloning and characterization of CXIP1, a novel PICOT domain-containing Arabidopsis protein that associates with CAX1. J Biol Chem 278:6503-9
Pittman, Jon K; Hirschi, Kendal D (2003) Don't shoot the (second) messenger: endomembrane transporters and binding proteins modulate cytosolic Ca2+ levels. Curr Opin Plant Biol 6:257-62
Hirschi, Kendal D (2003) Insertional mutants: a foundation for assessing gene function. Trends Plant Sci 8:205-7
Cheng, Ning-Hui; Pittman, Jon K; Barkla, Bronwyn J et al. (2003) The Arabidopsis cax1 mutant exhibits impaired ion homeostasis, development, and hormonal responses and reveals interplay among vacuolar transporters. Plant Cell 15:347-64
Hirschi, Karen K; Burt, Janis M; Hirschi, Kendal D et al. (2003) Gap junction communication mediates transforming growth factor-beta activation and endothelial-induced mural cell differentiation. Circ Res 93:429-37
Shigaki, Toshiro; Hirschi, Kendal D (2002) Chimeric gene construction without reference to restriction sites. Biotechniques 32:736, 738, 740
Gaxiola, Roberto A; Fink, Gerald R; Hirschi, Kendal D (2002) Genetic manipulation of vacuolar proton pumps and transporters. Plant Physiol 129:967-73

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