Plant natriuretic peptide active site determination and effects on cGMP and cell volume regulation
Yu Hua Wang A , Chris Gehring B , David M. Cahill C and Helen R. Irving A DA Department of Pharmaceutical Biology, Victorian College of Pharmacy, Monash University, 381 Royal Parade, Parkville, Vic 3052, Australia.
B Department of Biotechnology, University of the Western Cape, Private Bag X17, Bellville 7535, South Africa.
C School of Life and Environmental Sciences, Deakin University, Waurn Ponds, Vic 3217, Australia.
D Corresponding author. Email: helen.irving@vcp.monash.edu.au
Functional Plant Biology 34(7) 645-653 https://doi.org/10.1071/FP06316
Submitted: 1 December 2006 Accepted: 20 March 2007 Published: 4 July 2007
Abstract
Natriuretic peptides (NP) were first identified in animals where they play a role in the regulation of salt and water balance. This regulation is partly mediated by intracellular changes in cyclic GMP (cGMP). NP immunoanalogues occur in many plants and have been isolated, with two NP encoding genes characterised in Arabidopsis thaliana L. (AtPNP-A and AtPNP-B). Part of AtPNP-A contains the region with homology to human atrial (A)NP. We report here on the effects of recombinant AtPNP-A and smaller synthetic peptides within the ANP-homologous region with a view to identifying the biologically active domain of the molecule. Furthermore, we investigated interactions between AtPNP-A and the hormone, abscisic acid (ABA). ABA does not significantly affect Arabidopsis mesophyll protoplast volume regulation, whereas AtPNP-A and synthetic peptides promote water uptake into the protoplasts causing swelling. This effect is promoted by the membrane permeable cGMP analogue, 8-Br-cGMP, and inhibited by guanylate cyclase inhibitors indicating that increases in cGMP are an essential component of the plant natriuretic peptides (PNP) signalling cascade. ABA does not induce cGMP transients and does not affect AtPNP-A dependent cGMP increases, hence the two regulators differ in their second messenger signatures. Interestingly, AtPNP-A significantly delays and reduces the extent of ABA stimulated stomatal closure that is also based on cell volume regulation. We conclude that a complex interplay between observed PNP effects (stomatal opening and protoplast swelling) and ABA is likely to be cell type specific.
Additional keywords: abscisic acid, protoplast volume, stomata.
Acknowledgements
The authors wish to thank Dr M. Morse (University of the Western Cape) for the supply of the plasmid containing AtPNP-A and Dr P. Thompson (Monash University) for synthesising the peptides. Y. H. W. is supported by an Australian Postgraduate Award and this project is supported by the National Research Foundation of South Africa and the Australian Research Council’s Discovery funding scheme (project number DP0557561).
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