RESUMO
Plasma membrane (PM) proton-translocating adenosine triphosphatase (H+-ATPase) is a pivotal enzyme for plant growth and development that acts as a primary transporter and is activated by phosphorylation of the penultimate residue, threonine, at the C-terminus. Small Auxin-Up RNA family proteins maintain the phosphorylation level via inhibiting dephosphorylation of the residue by protein phosphatase 2C-D clade. Photosynthetically active radiation activates PM H+-ATPase via phosphorylation in mesophyll cells of Arabidopsis thaliana, and phosphorylation of PM H+-ATPase depends on photosynthesis and photosynthesis-related sugar supplementation, such as sucrose, fructose and glucose. However, the molecular mechanism and physiological role of photosynthesis-dependent PM H+-ATPase activation are still unknown. Analysis using sugar analogs, such as palatinose, turanose and 2-deoxy glucose, revealed that sucrose metabolites and products of glycolysis such as pyruvate induce phosphorylation of PM H+-ATPase. Transcriptome analysis showed that the novel isoform of the Small Auxin-Up RNA genes, SAUR30, is upregulated in a light- and sucrose-dependent manner. Time-course analyses of sucrose supplementation showed that the phosphorylation level of PM H+-ATPase increased within 10 min, but the expression level of SAUR30 increased later than 10 min. The results suggest that two temporal regulations may participate in the regulation of PM H+-ATPase. Interestingly, a 15NO3- uptake assay in leaves showed that light increases 15NO3- uptake and that increment of 15NO3- uptake depends on PM H+-ATPase activity. The results opened the possibility of the physiological role of photosynthesis-dependent PM H+-ATPase activation in the uptake of NO3-. We speculate that PM H+-ATPase may connect photosynthesis and nitrogen metabolism in leaves.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Nitratos/metabolismo , Fotossíntese , ATPases Translocadoras de Prótons/metabolismo , Folhas de Planta/metabolismo , Membrana Celular/metabolismo , Proteínas de Arabidopsis/metabolismo , Ácidos Indolacéticos/metabolismo , RNA/metabolismo , Açúcares/metabolismo , Sacarose/metabolismo , Glucose/metabolismoRESUMO
Plasma membrane (PM) H+-ATPase is crucial for light-induced stomatal opening and phosphorylation of a penultimate residue, Thr948 (pen-Thr, numbering according to Arabidopsis AHA1) is required for enzyme activation. In this study, a comprehensive phosphoproteomic analysis using guard cell protoplasts from Vicia faba shows that both red and blue light increase the phosphorylation of Thr881, of PM H+-ATPase. Light-induced stomatal opening and the blue light-induced increase in stomatal conductance are reduced in transgenic Arabidopsis plants expressing mutant AHA1-T881A in aha1-9, whereas the blue light-induced phosphorylation of pen-Thr is unaffected. Auxin and photosynthetically active radiation induce the phosphorylation of both Thr881 and pen-Thr in etiolated seedlings and leaves, respectively. The dephosphorylation of phosphorylated Thr881 and pen-Thr are mediated by type 2 C protein phosphatase clade D isoforms. Taken together, Thr881 phosphorylation, in addition of the pen-Thr phosphorylation, are important for PM H+-ATPase function during physiological responses, such as light-induced stomatal opening in Arabidopsis thaliana.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Fosforilação , Luz , Estômatos de Plantas/metabolismo , ATPases Translocadoras de Prótons/genética , ATPases Translocadoras de Prótons/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Membrana Celular/metabolismoRESUMO
In plants, cytosolic and extracellular pH homeostasis are crucial for various physiological processes, including the uptake of macronutrients and micronutrients, cell elongation, cell expansion, and enzyme activity. Proton (H+) gradients and the membrane potential are generated by a H+ pump consisting of an active primary transporter. Plasma membrane (PM) H+-ATPase, a PM-localized H+ pump, plays a pivotal role in maintaining pH homeostasis in plant cells and extracellular regions. PM H+-ATPase activity is regulated by protein abundance and by post-translational modifications. Several stimuli have been found to activate the PM H+-ATPase through phosphorylation of the penultimate threonine (Thr) of the carboxy terminus. Light- and photosynthesis-induced phosphorylation of PM H+-ATPase are conserved phenomena among various plant species. In this work, we review recent findings related to PM H+-ATPase regulation in the photosynthetic tissues of plants, focusing on its mechanisms and physiological roles. The physiological roles of photosynthesis-dependent PM H+-ATPase activation are discussed in the context of nitrate uptake and cytoplasmic streaming in leaves.