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Identification of plant vacuolar transporters mediating phosphate storage

ABSTRACT: Plant vacuoles serve as the primary intracellular compartments for inorganic phosphate (Pi) storage. Passage of Pi across vacuolar membranes plays a critical role in buffering the cytoplasmic Pi level against fluctuations of external Pi and metabolic activities. Here we demonstrate that the SPX-MFS proteins, designated as Phosphate Transporter 5 family (PHT5), also named Vacuolar Phosphate Transporter (VPT), function as vacuolar Pi transporters. Based on 31P-magnetic resonance spectroscopy analysis, Arabidopsis pht5;1 loss-of-function mutants accumulate less Pi and exhibit a lower vacuolar-to-cytoplasmic Pi ratio than controls. Conversely, overexpression of PHT5 leads to massive Pi sequestration into vacuoles and altered regulation of Pi starvation-responsive genes. Furthermore, we show that heterologous expression of OsSPX-MFS1, the rice PHT5 homolog, mediates Pi influx to yeast vacuoles. Our findings uncover a group of Pi transporters in vacuolar membranes that regulate the cytoplasmic Pi homeostasis required for the fitness of plant growth.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE74856 | GEO | 2016/03/28

SECONDARY ACCESSION(S): PRJNA301684

REPOSITORIES: GEO

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Identification of plant vacuolar transporters mediating phosphate storage

Project description:Plant vacuoles serve as the primary intracellular compartments for inorganic phosphate (Pi) storage. Passage of Pi across vacuolar membranes plays a critical role in buffering the cytoplasmic Pi level against fluctuations of external Pi and metabolic activities. Here we demonstrate that the SPX-MFS proteins, designated as Phosphate Transporter 5 family (PHT5), also named Vacuolar Phosphate Transporter (VPT), function as vacuolar Pi transporters. Based on 31P-magnetic resonance spectroscopy analysis, Arabidopsis pht5;1 loss-of-function mutants accumulate less Pi and exhibit a lower vacuolar-to-cytoplasmic Pi ratio than controls. Conversely, overexpression of PHT5 leads to massive Pi sequestration into vacuoles and altered regulation of Pi starvation-responsive genes. Furthermore, we show that heterologous expression of OsSPX-MFS1, the rice PHT5 homolog, mediates Pi influx to yeast vacuoles. Our findings uncover a group of Pi transporters in vacuolar membranes that regulate the cytoplasmic Pi homeostasis required for the fitness of plant growth. 10-day seedlings grown on Pi-sufficient medium or followed by 1-day or 3-day Pi starvation, shoot RNA and root RNA were extracted separately

2016-03-28 | E-GEOD-74856 | biostudies-arrayexpress

The proteome and phosphoproteome uncovers candidate proteins associated with vacuolar phosphate signal multipled by Vacuolar phosphate transporter1 (VPT1) in Arabidopsis

Project description:Plant vacuoles serve as the primary intracellular compartments for inorganic phosphate (Pi) storage. Passage of Pi across vacuolar membranes plays a critical role in buffering the cytoplasmic Pi level against fluctuations of external Pi and metabolic activities. To gain new insights into the proteins and processes vacuolar Pi level regulated by Vacuolar phosphate transporter1 (VPT1) in Arabidopsis, we carried out TMT labeling proteome and phosphoproteome profiling of Arabidopsis wild-type (WT) and vpt1 loss-of-function mutant plants. The vpt1 mutant had a reduced vacuolar Pi level, but an increased cytosol Pi level. The mutant was stunted as reflected in the reduction of the fresh weight compared with WT plants, and bolting earlier under normal growth conditions in soil. Over 5566 proteins and 7965 phosphopeptides were quantified. About 146 and 83 proteins were significantly changed at protein abundance or site-specific phosphorylation levels, but only 6 proteins were shared between them. Functional enrichment analysis revealed that Pi starvation signal in Arabidopsis vacuole is associated with photosynthesis, translation, RNA splicing, and defense response, consistent with similar studies in Arabidopsis. Except for PAP26, EIN2, and KIN10, which are reported to be associated with phosphate starvation signal, we also found many differential proteins involved in abscisic acid (ABA) signaling, such as CARK1, SnRK1, and AREB3, were changed in response to vacuolar Pi starvation. Our study illuminates several new aspects of the phosphate starvation response and identifies important targets for further investigation and potential crop improvement.

2023-03-08 | PXD040698 | iProX

Loss of arginine methylation in Saccharomyces cerevisiae leads to the dysregulation of phosphate-associated genes and processes

Project description:Hmt1p is the predominant arginine methyltransferase in Saccharomyces cerevisiae. Its substrate proteins are involved in transcription, transcriptional regulation, nucleocytoplasmic transport and RNA splicing. Functionally, Hmt1p-catalysed methylation can also modulate protein-protein interactions. Despite Hmt1p being well-characterised, the effects of its knockout on the proteome and transcriptome have not been reported. SILAC-based analyses of the hmt1Δ proteome, in mid-log exponential growth, revealed a decreased abundance of phosphate-associated proteins including Pho84p (phosphate transporter), Pho8p (vacuolar alkaline phosphatase), Pho3p (acid phosphatase) along with Vtc1p, Vtc3p and Vtc4p (subunits of the vacuolar transporter chaperone complex). RNA-Seq and microarray analysis revealed a downregulation of phosphate-responsive genes in hmt1Δ, including PHO5, PHO11 and PHO12 (acid phosphatases), PHO84 and PHO89 (phosphate transporters) and VTC3 (vacuolar transporter chaperone). Consistent with these observations, we observed a dysregulation of phosphate homeostasis in hmt1Δ, with a general decrease in extracellular phosphatase production and a decrease in total Pi in phosphate replete medium. We show that the transcription factor Pho4p, responsible for activation of the PHO pathway, can be methylated by Hmt1p at Arg-241 and is the likely cause of phosphate dysregulation in hmt1Δ. However, the methylation of Pho4p does not affect its nucleocytoplasmic localisation. We propose that the methylation of Pho4p may affect either its capacity to multimerise, its capacity to interact with Pho2p or target DNA, or may affect Pho4p phosphorylation at Ser-242 and/or Ser 243. Our study highlights a previously unknown function of Hmt1p in the regulation of phosphate homeostasis and suggests a means by which sensing of AdoMet may affect intracellular phosphate concentration.

2018-12-26 | GSE99869 | GEO

A. fumigatus wild type and delta phoB(PHO80) strains transcriptome analysis for growth upon minimal medium 11 mM Pi

Project description:Phosphate is an essential ion for fungal growth, required for proper DNA and phospholipids biosyntesis, energetic metabolism and signal transduction. The systems for inorganic phosphate (Pi) acquisition in eukaryotic cells (PHO) have been characterized as a low-affinity (that assures a supply of Pi at normal or high external Pi concentrations) and a high-affinity (activated in response to Pi starvation). Here, as an initial step to understand the PHO pathway in A. fumigatus, we characterized the PHO80 homologue, phoB(PHO80). We showed that the delta phoB(PHO80) mutant has a severe polar growth defect and there is an interaction between PhoB(PHO80), calcineurin and calcium metabolism. Microarray hybridizations carried out with RNA obtained from wild type and delta phoB(PHO80) mutant cells showed that Afu4g03610 [phoD(PHO84)], Afu7g06350 [phoE9(PHO89)], Afu4g06020 [phoCPHO81)], and Afu2g09040 (vacuolar transporter Vtc4) were more expressed either in the delta phoB(PHO80) mutant background or in 11 mM Pi. Keywords: gene expression array-based (log2 ratio)

2007-11-09 | GSE9550 | GEO

A. fumigatus wild type and delta phoB(PHO80) strains transcriptome analysis for growth upon minimal medium 0.1 mM Pi

Project description:Phosphate is an essential ion for fungal growth, required for proper DNA and phospholipids biosyntesis, energetic metabolism and signal transduction. The systems for inorganic phosphate (Pi) acquisition in eukaryotic cells (PHO) have been characterized as a low-affinity (that assures a supply of Pi at normal or high external Pi concentrations) and a high-affinity (activated in response to Pi starvation). Here, as an initial step to understand the PHO pathway in A. fumigatus, we characterized the PHO80 homologue, phoB(PHO80). We showed that the delta phoB(PHO80) mutant has a severe polar growth defect and there is an interaction between PhoB(PHO80), calcineurin and calcium metabolism. Microarray hybridizations carried out with RNA obtained from wild type and delta phoB(PHO80) mutant cells showed that Afu4g03610 [phoD(PHO84)], Afu7g06350 [phoE9(PHO89)], Afu4g06020 [phoCPHO81)], and Afu2g09040 (vacuolar transporter Vtc4) were more expressed either in the delta phoB(PHO80) mutant background or in 0.1 mM Pi. Keywords: gene expression array-based (log2 ratio)

2007-11-09 | GSE9548 | GEO

Arabidopsis thaliana

Project description:Identification of plant vacuolar transporters mediating phosphate storage

| PRJNA301684 | ENA

Gene expression analysis on Hydrangea macrophylla sepal pigmentation due to aluminum accumulation

Project description:Most blue color in flowers is due to anthocyanin, and considerable proportion of blue coloration can be attributed to metal-complexed anthocyanins. Recently, we reported vacuolar localized iron-transporter in blue petal cells of Tulipa gesneriana. However the mechanism of another metal ion transporters and subsequent flower color development has yet to be fully explored. In Hydrangea macrophylla, Al3+ is involved in blue coloration and the anthocyanin is formed Al3+-complex in vacuoles. To identify the molecular mechanism of blue coloration in hydrangea flowers, we tried to isolate the related genes transporting metal ion into vacuoles. From the sepal cDNA library we read the sequences of ca. 12000 genes, then a microarray analysis was carried out. From the sequences information, we chose several genes that might localize vacuolar membrane and transport Al3+. By using Al3+-sensitive yeast strain, we could identify the gene transporting Al3+ into vacuole. From the functional similarity and predicted localization, we could also identify the gene transporting Al3+ into cytosol. We will report the Al3+ mobilization from out of cell into vacuole in the sepal of Hydrangea macrophylla.

2012-12-20 | GSE26665 | GEO

Genome wide transcriptomic analysis reveals the mechanism underlying Stylosanthes roots in response to phosphorus deficiency

Project description:Phosphorus (P) is an important macronutrient for plant growth that participates in a series of biological processes. Thus, P deficiency is considered as one of the major constraints limiting crop growth and yield. Although stylo (Stylosanthes) is a dominate tropical legume that displays adaptation to low phosphate (Pi) availability in acid soils, its adaptive mechanisms remain largely unknown. In this study, variation in low P stress tolerance was investigated using two stylo cultivars in hydroponics, which revealed that the stylo RY2 cultivar exhibited higher adaptability to Pi starvation than the RY5 cultivar, as reflected by less reduction in dry weight under P deficient condition, and accompanied by higher P concentrations in shoot and root. Furthermore, better root growth performance and higher APase activity were found in leaves and roots of stylo RY2 cultivar compared to RY5. RNA‐seq analysis revealed 8,348 genes that exhibited differentially expressed under P deficient and sufficient conditions in RY2 roots, with many Pi starvation up-regulated genes associated with P metabolic process, protein modification process, transport and other metabolic processes. A group of differentially expressed genes (DEGs) involved in Pi uptake and homeostasis were identified, such as genes encoding Pi transporter (PT), purple acid phosphatase (PAP), multidrug and toxin extrusion (MATE) and aluminum-activated malate transporter (ALMT). Furthermore, a variety of genes related to transcription factors (TFs) and regulators involved in Pi signaling, including genes belonging to the PHOSPHATE STARVATION RESPONSE 1-like (PHR1), WRKY family, the SYG1/PHO81/XPR1 (SPX) domain, bHLH and ARP family, were also regulated by P deficiency in stylo roots. Taken together, this study suggests a complex adaptive response of stylo to P deficiency, contributing to maintain Pi homeostasis.

2022-12-31 | GSE171448 | GEO

Transcriptome sequencing of WT and inorganic phosphate transporter (TgPiT) ortholog-deficient Toxoplasma parasites

Project description:Purpose: Toxoplasma must import inorganic phosphate via a transporter at the plasma membrane. Methods: WT and TgPiT-deficient parasites were grown in human foreskin fibroblasts for 2 days. The parasites were syringed, filter-purified, and resuspended n ice-cold PBS. A trizol-based method was used to extract total RNA from both strains. Toral RNA was subjected to cDNA library construction using Ilumina. Results: The parasite expresses a unique TgPiT as selective Pi transporter, located at the plasma membrane and on VAC internal membranes of endosomal organelles. Conclusion: TgPiT is essential for parasite survival, virulence, ionic homeostasis and osmoregulation

2021-03-01 | GSE145660 | GEO

Renal Fanconi syndrome and hypoposphatemic rickets in mice deficient for retroviral receptor XPR1 in the nephron

Project description:Tight control of both extracellular and intracellular inorganic phosphate (Pi) levels is critical to the normal functioning of virtually all biochemical and physiological processes. The kidney participates in Pi homeostasis by controlling Pi reabsorption from the primary urine. Pi is freely filtered at the kidney glomerulus and is reabsorbed in the renal tubule by the action of the apical sodium-dependent phosphate transporters NaPi-IIa/NaPi-IIc/Pit2. The molecular identity of transporter(s) involved in the basolateral Pi efflux remains unknown. Recent evidence has suggested that the retroviral receptor XPR1 might be a candidate for this role. Here we show that conditional inactivation of Xpr1 in the renal tubule in mice results in impaired renal Pi reabsorption associated with a generalized proximal tubular dysfunction, or Fanconi syndrome, characterized by glycosuria, aminoaciduria, calciuria and albuminuria. Bone histomorphometry showed that the Xpr1-deficient mice develop hypophosphatemic osteomalacia secondary to the renal dysfunction. The analysis of Pi transport in primary culture of the proximal tubular cells revealed that the Pi efflux was significantly affected in cells devoid of Xpr1. These results identify XPR1 as a major player in Pi homeostasis and as a potential therapeutic target in bone and kidney disorders.

2016-12-01 | GSE87450 | GEO

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