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Summary: In grapevine, climate changes lead to increased berry potassium (K+) contents that result in must with low acidity. Consequently, wines are becoming 'flat' to the taste, with poor organoleptic properties and low potential aging, resulting in significant economic loss. Precise investigation into the molecular determinants controlling berry K+ accumulation during its development are only now emerging.Here, we report functional characterization by electrophysiology of a new grapevine Shaker‐type K+ channel, VvK3.1. The analysis of VvK3.1 expression patterns was performed by qPCR and in situ hybridization.We found that VvK3.1 belongs to the AKT2 channel phylogenetic branch and is a weakly rectifying channel, mediating both inward and outward K+ currents. We showed that VvK3.1 is highly expressed in the phloem and in a unique structure located at the two ends of the petiole, identified as a pulvinus.From the onset of fruit ripening, all data support the role of the VvK3.1 channel in the massive K+ fluxes from the phloem cell cytosol to the berry apoplast during berry K+ loading. Moreover, the high amount of VvK3.1 transcripts detected in the pulvinus strongly suggests a role for this Shaker in the swelling and shrinking of motor cells involved in paraheliotropic leaf movements. [ABSTRACT FROM AUTHOR]

1: In the human airway epithelium, VIP/PACAP receptors are distributed in nerve fibers and in epithelial cells but their role in transepithelial ion transport have not been reported. Here, we show that human bronchial epithelial Calu-3 cells expressed the VPAC1 receptor subtype which shares similar high affinity for VIP and PACAP-27. 2: The stoichiometric binding parameters characterizing the 125I-VIP and 125I-PACAP-27 binding to these receptors were determined. 3: We found that VIP (EC50˜7.6?nM) and PACAP-27 (EC50˜10?nM) stimulated glibenclamide-sensitive and DIDS-insensitive iodide efflux in Calu-3 cells. 4: The protein kinase A (PKA) inhibitor, H-89 and the protein kinase C (PKC) inhibitor, chelerythrine chloride prevented activation by both peptides demonstrating that PKA and PKC are part of the signaling pathway. This profile corresponds to the pharmacological signature of CFTR. 5: In the cystic fibrosis airway epithelial IB3-1 cell lacking functional CFTR but expressing VPAC1 receptors, neither VIP, PACAP-27 nor forskolin stimulated chloride transport. 6: Ussing chamber experiments demonstrated stimulation of CFTR-dependent short-circuit currents by VIP or PACAP-27 applied to the basolateral but not to the apical side of Calu-3 cells monolayers. 7: This study shows the stimulation in human bronchial epithelial cells of CFTR-dependent chloride secretion following activation by VIP and PACAP-27 of basolateral VPAC1 receptors.British Journal of Pharmacology (2004) 141, 698-708. doi:10.1038/sj.bjp.0705597 [ABSTRACT FROM AUTHOR]

The regulation of the GORK (Guard Cell Outward Rectifying) Shaker channel mediating a massive K+ efflux in Arabidopsis guard cells by the phosphatase AtPP2CA was investigated. Unlike the gork mutant, the atpp2ca mutants displayed a phenotype of reduced transpiration. We found that AtPP2CA interacts physically with GORK and inhibits GORK activity in Xenopus oocytes. Several amino acid substitutions in the AtPP2CA active site, including the dominant interfering G145D mutation, disrupted the GORK-AtPP2CA interaction, meaning that the native conformation of the AtPP2CA active site is required for theGORKAtPP2CA interaction. Furthermore, two serines in the GORK ankyrin domain that mimic phosphorylation (Ser to Glu) or dephosphorylation (Ser to Ala) were mutated. Mutations mimicking phosphorylation led to a significant increase in GORK activity, whereas mutations mimicking dephosphorylation had no effect on GORK. In Xenopus oocytes, the interaction of AtPP2CA with "phosphorylated" or "dephosphorylated"GORK systematically led to inhibition of the channel to the same baseline level. Single-channel recordings indicated that the GORK S722E mutation increases the open probability of the channel in the absence, but not in the presence, of AtPP2CA. The dephosphorylation- independent inactivation mechanism of GORK by AtPP2CA is discussed in relation with well known conformational changes in animal Shaker-like channels that lead to channel opening and closing. In plants, PP2C activity would control the stomatal aperture by regulating both GORK and SLAC1, the two main channels required for stomatal closure. [ABSTRACT FROM AUTHOR]

In grapevine, climate changes lead to increased berry potassium (K + ) contents that result in must with low acidity. Consequently, wines are becoming 'flat' to the taste, with poor organoleptic properties and low potential aging, resulting in significant economic loss. Precise investigation into the molecular determinants controlling berry K + accumulation during its development are only now emerging. Here, we report functional characterization by electrophysiology of a new grapevine Shaker-type K + channel, VvK3.1. The analysis of VvK3.1 expression patterns was performed by qPCR and in situ hybridization. We found that VvK3.1 belongs to the AKT2 channel phylogenetic branch and is a weakly rectifying channel, mediating both inward and outward K + currents. We showed that VvK3.1 is highly expressed in the phloem and in a unique structure located at the two ends of the petiole, identified as a pulvinus. From the onset of fruit ripening, all data support the role of the VvK3.1 channel in the massive K + fluxes from the phloem cell cytosol to the berry apoplast during berry K + loading. Moreover, the high amount of VvK3.1 transcripts detected in the pulvinus strongly suggests a role for this Shaker in the swelling and shrinking of motor cells involved in paraheliotropic leaf movements., (© 2018 The Authors. New Phytologist © 2018 New Phytologist Trust.)

The combined daily consumption of fresh water ranges from 200 to 700 liters per capita per day in most developed countries, with about 70% being used for agricultural needs. Unlike other resources such as the different forms of energy, water has no other alternatives. With the looming prospect of global water crisis, the recent laudable success in deciphering the early steps in the signal transduction of the "stress hormone" abscisic acid (ABA) has ignited hopes that crops can be engineered with the capacity to maintain productivity while requiring less water input. Although ABA was first discovered in plants, it has resurfaced in the human brain (and many other non-plant organisms : sea sponge, some parasites, hydra to name a few), suggesting that its existence may be widespread. In humans, more amazingly, ABA has shown anti-inflammatory and antiviral properties. Even its receptors and key signaling intermediates have homologs in the human genome suggesting that evolution has re-fashioned these same proteins into new functional contexts. Thus, learning about the molecular mechanisms of ABA in action using the more flexible plant model will be likely beneficial to other organisms, and especially in human diseases, which is topical in the medical circle. ABA can accumulate up to 10 to 30-fold in plants under drought stress relative to unstressed conditions. The built up of the hormone then triggers diverse adaptive pathways permitting plants to withstand temporary bouts of water shortage. One favorite experimental model to unravel ABA signaling mechanisms in all of its intimate detail is based on the hormone's ability to elicit stomatal closure - a rapid cellular response of land plants to limit water loss through transpiration. Each microscopic stoma, or pore, is contoured by two specialized kidney-shaped cells called the guard cells. Because land plants are protected by a waxy cuticle impermeable to gas exchange, the stomatal pores are thus the primary portals for photosynthetic CO(2) uptake. Drought, by biasing pathways that lead to rapid closure of these pores, has therefore a negative impact on photosynthesis, and consequently, biomass as well. The stomatal aperture widens and narrows by expansion and contraction, respectively, of these flanking guard cells caused by changes in the intracellular concentrations of ion fluxes. These transport mechanisms most likely share fundamental principles with any excitable cell. These events require coordination of channels, vacuolar and membrane transporters that generate a specific pattern of electrical signals that relay the ABA stimulus. Research on ABA begun in the 1960's has now been crowned by the achievement of having identified the soluble ABA receptor that turns on and off the activities of a kinase/phosphatase pair, as the heart of the signaling complex. Results distilled from the latest structural studies on these ABA receptors, characterized by the so-called START domain, are beginning to tender the most exciting promise for rational design of agonists and antagonists towards modulating stress adaptive ability in plants. This review will chart the recent extraordinary progress that has enlightened us on how ABA controls membrane transport mechanisms that evoke the fast stomatal closing pathway., (© Société de Biologie, 2013.)

1. In the human airway epithelium, VIP/PACAP receptors are distributed in nerve fibers and in epithelial cells but their role in transepithelial ion transport have not been reported. Here, we show that human bronchial epithelial Calu-3 cells expressed the VPAC(1) receptor subtype which shares similar high affinity for VIP and PACAP-27. 2. The stoichiometric binding parameters characterizing the (125)I-VIP and (125)I-PACAP-27 binding to these receptors were determined. 3. We found that VIP (EC(50) approximately 7.6 nM) and PACAP-27 (EC(50) approximately 10 nM) stimulated glibenclamide-sensitive and DIDS-insensitive iodide efflux in Calu-3 cells. 4. The protein kinase A (PKA) inhibitor, H-89 and the protein kinase C (PKC) inhibitor, chelerythrine chloride prevented activation by both peptides demonstrating that PKA and PKC are part of the signaling pathway. This profile corresponds to the pharmacological signature of CFTR. 5. In the cystic fibrosis airway epithelial IB3-1 cell lacking functional CFTR but expressing VPAC(1) receptors, neither VIP, PACAP-27 nor forskolin stimulated chloride transport. 6. Ussing chamber experiments demonstrated stimulation of CFTR-dependent short-circuit currents by VIP or PACAP-27 applied to the basolateral but not to the apical side of Calu-3 cells monolayers. 7. This study shows the stimulation in human bronchial epithelial cells of CFTR-dependent chloride secretion following activation by VIP and PACAP-27 of basolateral VPAC(1) receptors.