Expression Of Plasma Membrane Research Articles

Apple vacuolar sugar transporters regulated by MdDREB2A enhance drought resistance by promoting accumulation of soluble sugars and activating ABA signaling

Abstract Soluble sugars are not only an important contributor to fruit quality, but also serve as the osmotic regulators in response to abiotic stresses. Early drought stress promotes sugar accumulation, while specific sugar transporters govern the cellular distribution of the sugars. Here, we show that apple plantlets accumulate soluble sugars in leaf tissues under drought stress. Transcriptional profiling of stressed and control plantlets revealed differential expression of several plasma membrane- or vacuolar membrane-localized sugar transporter genes. Among these, four previously identified vacuolar sugar transporter (VST) genes (MdERDL6–1, MdERDL6–2, MdTST1 and MdTST2) showed higher expression under drought, suggesting their roles in response to drought stress. Promoter cis-elements analyses, yeast one-hybrid and dual-luciferase tests confirmed that the drought-induced transcription factor MdDREB2A could promote the expression of MdERDL6–1/−2 and MdTST1/2 by binding to their promoter regions. Moreover, overexpressing of each of these four MdVSTs alone in transgenic apple or Arabidopsis plants accumulated more soluble sugars and abscisic acid, and enhanced drought resistance. Furthermore, apple plants overexpressing MdERDL6–1 also showed reduced water potential, facilitated stomatal closure and reactive oxygen species scavenging under drought condition compared to control plants. Overall, our results suggest a potential strategy to enhance drought resistance and sugar accumulation in fruits through manipulating the genes involved in vacuolar sugar transport.

The asymmetric expression of plasma membrane H+-ATPase family genes in response to pulvinus-driven leaf phototropism movement in Vitis vinifera.

Phototropism movement is crucial for plants to adapt to various environmental changes. Plant P-type H+-ATPase (HA) plays diverse roles in signal transduction during cell expansion, regulation of cellular osmotic potential and stomatal opening, and circadian movement. Despite numerous studies on the genome-wide analysis of Vitis vinifera, no research has been done on the P-type H+-ATPase family genes, especially concerning pulvinus-driven leaf movement. In this study, 55 VvHAs were identified and classified into nine distinct subgroups (1 to 9). Gene members within the same subgroups exhibit similar features in motif, intron/exon, and protein tertiary structures. Furthermore, four pairs of genes were derived by segmental duplication in grapes. Cis-acting element analysis identified numerous light/circadian-related elements in the promoters of VvHAs. qRT-PCR analysis showed that several genes of subgroup 7 were highly expressed in leaves and pulvinus during leaf movement, especially VvHA14, VvHA15, VvHA16, VvHA19, VvHA51, VvHA52, and VvHA54. Additionally, we also found that the VvHAs genes were asymmetrically expressed on both sides of the extensor and flexor cell of the motor organ, the pulvinus. The expression of VvHAs family genes in extensor cells was significantly higher than that in flexor cells. Overall, this study serves as a foundation for further investigations into the functions of VvHAs and contributes to the complex mechanisms underlying grapevine pulvinus growth and development.

GreenGate 2.0: Backwards compatible addons for assembly of complex transcriptional units and their stacking with GreenGate.

Molecular cloning is a crucial technique in genetic engineering that enables the precise design of synthetic transcriptional units (TUs) and the manipulation of genomes. GreenGate and several other modular molecular cloning systems were developed about ten years ago and are widely used in plant research. All these systems define grammars for assembling transcriptional units from building blocks, cloned as Level 0 modules flanked by four-base pair overhangs and recognition sites for a particular Type IIs endonuclease. Modules are efficiently assembled into Level 1 TUs in a hierarchical assembly process, and Level 2 multigene constructs are assembled by stacking Level 1 TUs. GreenGate is highly popular but has three main limitations. First, using ad-hoc overhangs added by PCR and classical restriction/ligation prevents the efficient use of a one-pot, one-step reaction to generate entry clones and domesticate internal sites; second, a Level 1 TU is assembled from a maximum of six modules, which may be limiting for applications such as multiplex genome editing; third, the generation of Level 2 assemblies is sequential and inefficient. GreenGate 2.0 (GG2.0) expands GreenGate features. It introduces additional overhangs, allowing for the combination of up to 12 Level 0 modules in a Level 1 TU. It includes a Universal Entry Generator plasmid (pUEG) to streamline the generation of Level 0 modules. GG2.0 introduces GreenBraid, a convenient method for stacking transcriptional units iteratively for multigene assemblies. Importantly, GG2.0 is backwards compatible with most existing GreenGate modules. Additionally, GG2.0 includes Level 0 modules for multiplex expression of guide RNAs for CRISPR/Cas9 genome editing and pre-assembled Level 1 vectors for dexamethasone-inducible gene expression and ubiquitous expression of plasma membrane and nuclear fluorescent markers. GG2.0 streamlines and increases the versatility of assembling complex transcriptional units and their combination.

The Role of Ergosterol and Sphingolipids in the Localization and Activity of Candida albicans' Multidrug Transporter Cdr1p and Plasma Membrane ATPase Pma1p.

Opportunistic pathogen Candida albicans causes systemic infections named candidiasis. Due to the increasing number of multi-drug resistant clinical isolates of Candida sp., currently employed antifungals (e.g., azoles) are insufficient for combating fungal infection. One of the resistance mechanisms toward azoles is increased expression of plasma membrane (PM) transporters (e.g., Cdr1p), and such an effect was observed in C. albicans clinical isolates. At the same time, it has been proven that a decrease in PMs sphingolipids (SLs) content correlates with altered sensitivity to azoles and diminished Cdr1p levels. This indicates an important role for SL in maintaining the properties of PM and gaining resistance to antifungal agents. Here, we prove using a novel spot variation fluorescence correlation spectroscopy (svFCS) technique that CaCdr1p localizes in detergent resistant microdomains (DRMs). Immunoblot analysis confirmed the localization of CaCdr1p in DRMs fraction in both the C. albicans WT and erg11Δ/Δ strains after 14 and 24 h of culture. We also show that the C. albicans erg11Δ/Δ strain is more sensitive to the inhibitor of SLs synthesis; aureobasidin A (AbA). AbA treatment leads to a diminished amount of SLs in C. albicans WT and erg11Δ/Δ PM, while, for C. albicans erg11Δ/Δ, the general levels of mannose-inositol-P-ceramide and inositol-P-ceramide are significantly lower than for the C. albicans WT strain. Simultaneously, the level of ergosterol in the C. albicans WT strain after adding of AbA remains unchanged, compared to the control conditions. Analysis of PM permeabilization revealed that treatment with AbA correlates with the disruption of PM integrity in C. albicans erg11Δ/Δ but not in the C. albicans WT strain. Additionally, in the C. albicans WT strain, we observed lower activity of H+-ATPase, correlated with the delocalization of both CaCdr1p and CaPma1p.

Exogenous Application of Alpha-Lipoic Acid Mitigates Salt-Induced Oxidative Damage in Sorghum Plants through Regulation Growth, Leaf Pigments, Ionic Homeostasis, Antioxidant Enzymes, and Expression of Salt Stress Responsive Genes.

In plants, α-Lipoic acid (ALA) is considered a dithiol short-chain fatty acid with several strong antioxidative properties. To date, no data are conclusive regarding its effects as an exogenous application on salt stressed sorghum plants. In this study, we investigated the effect of 20 µM ALA as a foliar application on salt-stressed sorghum plants (0, 75 and 150 mM as NaCl). Under saline conditions, the applied-ALA significantly (p ≤ 0.05) stimulated plant growth, indicated by improving both fresh and dry shoot weights. A similar trend was observed in the photosynthetic pigments, including Chl a, Chl b and carotenoids. This improvement was associated with an obvious increase in the membrane stability index (MSI). At the same time, an obvious decrease in the salt induced oxidative damages was seen when the concentration of H2O2 and malondialdehyde (MDA) was reduced in the salt stressed leaf tissues. Generally, ALA-treated plants demonstrated higher antioxidant enzyme activity than in the ALA-untreated plants. A moderate level of salinity (75 mM) induced the highest activities of superoxide dismutase (SOD), guaiacol peroxidase (G-POX), and ascorbate peroxidase (APX). Meanwhile, the highest activity of catalase (CAT) was seen with 150 mM NaCl. Interestingly, applied-ALA led to a substantial decrease in the concentration of both Na and the Na/K ratio. In contrast, K and Ca exhibited a considerable increase in this respect. The role of ALA in the regulation of K+/Na+ selectivity under saline condition was confirmed through a molecular study (RT-PCR). It was found that ALA treatment downregulated the relative gene expression of plasma membrane (SOS1) and vacuolar (NHX1) Na+/H+ antiporters. In contrast, the high-affinity potassium transporter protein (HKT1) was upregulated.

Augmented CO2 tolerance by expressing a single H+-pump enables microalgal valorization of industrial flue gas

Microalgae can accumulate various carbon-neutral products, but their real-world applications are hindered by their CO2 susceptibility. Herein, the transcriptomic changes in a model microalga, Chlamydomonas reinhardtii, in a high-CO2 milieu (20%) are evaluated. The primary toxicity mechanism consists of aberrantly low expression of plasma membrane H+-ATPases (PMAs) accompanied by intracellular acidification. Our results demonstrate that the expression of a universally expressible PMA in wild-type strains makes them capable of not only thriving in acidity levels that they usually cannot survive but also exhibiting 3.2-fold increased photoautotrophic production against high CO2 via maintenance of a higher cytoplasmic pH. A proof-of-concept experiment involving cultivation with toxic flue gas (13 vol% CO2, 20 ppm NOX, and 32 ppm SOX) shows that the production of CO2-based bioproducts by the strain is doubled compared with that by the wild-type, implying that this strategy potentially enables the microalgal valorization of CO2 in industrial exhaust.

60P CCL2 expression and the neutrophil plasma membrane characteristics in blood in different stages of endometrial cancer

The mechanisms of neutrophils (Nph) recruitment to a tumor and their pro-tumor polarization are poorly understood. The CCL family of chemokines enhance the tumor recruitment, and the plasma membrane topography affects Nph activity. We aimed to evaluate the CCL2 expression, the stiffness and surface markers expression of Nph plasma membrane in blood in endometrial cancer (EC). Blood samples were obtained from the primary endometrioid EC patients, stage I-III FIGO (n=58), uterine myoma patients (n=20) and healthy women (n=30). The CCL2 level was determined by ELISA (JSC Vektor-Best-Volga, Russia). Circulating Nph plasma membrane stiffness was measured by atomic force microscopy (NT-MDT, Russia). The CD16, CD95 Nph surface expression was assessed by fluorescence microscopy. Statistical analysis was performed using Statistica 13. We observed that serum CCL2 level was decreased in myoma (197.6 (Q1-Q3 147.1-255.1) pg/ml) (p=0.013) and increased in stage I EC (299.6 (255.8-411.8) pg/ml) (p=0.012) compared with the control (270.1 (220.1-310.8) pg/ml). In stage II-III, serum CCL2 level was lower (236.9 (190.2-276.3) pg/ml) than that in stage I (p=0.007). The Nph membrane stiffness in stage I was reduced compared with the control (37.3±2.2 MPa vs 56.7±2.4 MPa) (p=0.051). In stage II-III, the membrane stiffness (70.5±8.4 MPa) was higher than in myoma (p=0.021) and the control (p=0.046). There was a linear regression relationship between serum CCL2 level and Nph membrane stiffness value in stage I (p=0.023), but not in stage II-III. The expression of CD95, a marker involved in immune evasion, was lower in stage II-III compared with myoma (p=0.003) and the control (p=0.006). In EC, membrane stiffness negatively correlated with CD95 expression (r=-0.478, p=0.0005). The number of active mature (CD16+) Nph was lower in stage I compared with myoma (p=0.029) and negatively correlated with serum CCL2 level (r=-0.423, p=0.0024). The serum CCL2 expression, the structure and marker status of circulating Nph membrane in stage I endometrioid EC differ from those in stage II-III, probably reflecting differences in the Nph recruitment and polarization processes.

Gibberellin biosynthesis inhibitor mepiquat chloride enhances root K+ uptake in cotton by modulating plasma membrane H+-ATPase.

Potassium deficiency causes severe losses in yield and quality in crops. Mepiquat chloride, a plant growth regulator, can increase K+ uptake in cotton (Gossypium hirsutum), but the underlying physiological mechanisms remain unclear. In this study, we used a non-invasive micro-test technique to measure K+ and H+ fluxes in the root apex with or without inhibitors of K+ channels, K+ transporters, non-selective cation channels, and plasma membrane H+-ATPases. We found that soaking seeds in mepiquat chloride solution increased the K+ influx mediated by K+ channels and reduced the K+ efflux mediated by non-selective cation channels in cotton seedlings. Mepiquat chloride also increased negative membrane potential (Em) and the activity of plasma membrane H+-ATPases in roots, due to higher levels of gene expression and protein accumulation of plasma membrane H+-ATPases as well as phosphorylation of H+-ATPase 11 (GhAHA11). Thus, plasma membrane hyperpolarization mediated by H+-ATPases was able to stimulate the activity of K+ channels in roots treated with mepiquat chloride. In addition, reduced K+ efflux under mepiquat chloride treatment was associated with reduced accumulation of H2O2 in roots. Our results provide important insights into the mechanisms of mepiquat chloride-induced K+ uptake in cotton and hence have the potential to help in improving K nutrition for enhancing cotton production.

Appropriate NH4 +/NO3 - Ratio Triggers Plant Growth and Nutrient Uptake of Flowering Chinese Cabbage by Optimizing the pH Value of Nutrient Solution.

Compared with sole nitrogen (N), the nutrition mixture of ammonium (NH4+) and nitrate (NO3–) is known to better improve crop yield and quality. However, the mechanism underlying this improvement remains unclear. In the present study, we analyzed the changes in nutrient solution composition, content of different N forms in plant tissues and exudates, and expression of plasma membrane (PM) H+-ATPase genes (HAs) under different NH4+/NO3– ratios (0/100, 10/90, 25/75, 50/50 as control, T1, T2, and T3) in flowering Chinese cabbage. We observed that compared with the control, T1 and T2 increased the economical yield of flowering Chinese cabbage by 1.26- and 1.54-fold, respectively, whereas T3 significantly reduced plant yield. Compared with the control, T1–T3 significantly reduced the NO3– content and increased the NH4+, amino acid, and soluble protein contents of flowering Chinese cabbage to varying extents. T2 significantly increased the N use efficiency (NUE), whereas T3 significantly decreased it to only being 70.25% of that of the control. Owing to the difference in N absorption and utilization among seedlings, the pH value of the nutrient solution differed under different NH4+/NO3– ratios. At harvest, the pH value of T2 was 5.8; in the control and T1, it was approximately 8.0, and in T3 it was only 3.6. We speculated that appropriate NH4+/NO3– ratios may improve N absorption and assimilation and thus promote the growth of flowering Chinese cabbage, owing to the suitable pH value. On the contrary, addition of excessive NH4+ may induce rhizosphere acidification and ammonia toxicity, causing plant growth inhibition. We further analyzed the transcription of PM H+-ATPase genes (HAs). HA1 and HA7 transcription in roots was significantly down-regulated by the addition of the mixture of NH4+ and NO3–, whereas the transcription of HA2, HA9 in roots and HA7, HA8, and HA10 in leaves was sharply up-regulated by the addition of the mixture; the transcription of HA3 was mainly enhanced by the highest ratio of NH4+/NO3–. Our results provide valuable information about the effects of treatments with different NH4+/NO3– ratios on plant growth and N uptake and utilization.

Overtime expression of plasma membrane and mitochondrial function markers associated with cell death in human spermatozoa exposed to nonphysiological levels of reactive oxygen species.

In many cell types, the potential of reactive oxygen species to induce death processes has been largely demonstrated. Studies in spermatozoa have associated the imbalance of reactive oxygen species and phosphatidylserine externalisation as an apoptosis marker. However, the lack of consensus about time effect in the joint expression of these and other death markers has made it difficult to understand the set of mechanisms influenced beyond the concentration effect of reactive oxygen species to stimulate cell death. Here, the plasma membrane permeability and integrity, phosphatidylserine externalisation and mitochondrial membrane potential were jointly evaluated as death markers in human spermatozoa stimulated with H2 O2 . The results showed a profound and sustained effect of dissipation in the mitochondrial membrane potential and an increased phosphatidylserine externalisation in human spermatozoa exposed to 3mmol-1 of H2 O2 at 30min. This was followed by an increased membrane permeability after 45min. The last observed event was the loss of cell membrane integrity at 60min. In conclusion, mitochondria are rapidly affected in human spermatozoa exposed to reactive oxygen species, with the barely detectable mitochondrial membrane potential coexisting with the high phosphatidylserine externalisation in cells with normal membrane permeability.

NO is involved in H2-induced adventitious rooting in cucumber by regulating the expression and interaction of plasma membrane H+-ATPase and 14-3-3.

NO was involved in H2-induced adventitious rooting by regulating the protein and gene expressions of PM H+-ATPase and 14-3-3. Simultaneously, the interaction of PM H+-ATPase and 14-3-3 protein was also involved in this process. Hydrogen gas (H2) and nitric oxide (NO) have been shown to be involved in plant growth and development. The results in this study revealed that NO was involved in H2-induced adventitious root formation. Western blot (WB) analysis showed that the protein abundances of plasma membrane H+-ATPase (PM H+-ATPase) and 14-3-3 protein were increased after H2, NO, H2 plus NO treatments, whereas their protein abundances were down regulated when NO scavenger carboxy-2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTI O) was added. Moreover, the mRNA abundances of the HA3 and 14-3-3(7) gene as well as the activities of PM H+-ATPase (EC 3.6.1.35) and H+ pump were in full agreement with the changes of protein abundance. Phosphorylation of PM H+-ATPase and the interaction of PM H+-ATPase and 14-3-3 protein were detected by co-immunoprecipitation analysis. H2 and NO significantly up regulated the phosphorylation of PM H+-ATPase and the interaction of PM H+-ATPase and 14-3-3 protein. Conversely, the stimulation of PM H+-ATPase phosphorylation and protein interaction were significantly diminished by cPTIO. Protein interaction activator fusicoccin (FC) and inhibitor adenosine monophosphate (AMP) of PM H+-ATPase and 14-3-3 were used in this study, and the results showed that FC significantly increased the abundances of PM H+-ATPase and 14-3-3, while AMP showed opposite trends. We further proved the critical roles of PM H+-ATPase and 14-3-3 protein interaction in NO-H2-induced adventitious root formation. Taken together, our results suggested that NO might be involved in H2-induced adventitious rooting by regulating the expression and the interaction of PM H+-ATPase and 14-3-3 protein.

Partial cloning, characterization, and analysis of expression and activity of plasma membrane H+-ATPase in Kallar grass [Leptochloa fusca (L.) Kunth] under salt stress.

Kallar grass (Leptochloa fusca) is a highly salt-tolerant C4 perennial halophytic forage. The regulation of ion movement across the plasma membrane (PM) to improve salinity tolerance of plant is thought to be accomplished with the aid of the proton electrochemical gradient generated by PM H+-ATPase. In this study, we cloned a partial gene sequence of the Lf PM H+-ATPase and investigated its expression and activity under salt stress. The amino acid sequence of the isolated region of Lf PM H+-ATPase possesses the maximum identity up to 96% to its ortholog in Aeluropus littoralis. The isolated fragment of Lf PM H+-ATPase gene is a member of the subfamily Π of plant PM H+-ATPase and is most closely related to the Oryza sativa gene OSA7. The transcript level and activity of the PM H+-ATPase were increased in roots and shoots in response to NaCl and were peaked at 450mM NaCl in both tissues. The induction of activity and gene expression of PM H+-ATPase in roots and shoots of Kallar grass under salinity indicate the necessity for this pump in these organs during salinity adaptation to establish and maintain the electrochemical gradient across the PM of the cells for adjusting ion homeostasis.

Tissue Tolerance Coupled With Ionic Discrimination Can Potentially Minimize the Energy Cost of Salinity Tolerance in Rice.

Salinity is one of the major constraints in rice production. To date, development of salt-tolerant rice cultivar is primarily focused on salt-exclusion strategies, which incur greater energy cost. The present study aimed to evaluate a balancing strategy of ionic discrimination vis-à-vis tissue tolerance, which could potentially minimize the energy cost of salt tolerance in rice. Four rice genotypes, viz., FL478, IR29, Kamini, and AC847, were grown hydroponically and subjected to salt stress equivalent to 12 dS m–1 at early vegetative stage. Different physiological observations (leaf chlorophyll content, chlorophyll fluorescence traits, and tissue Na+ and K+ content) and visual scoring suggested a superior Na+-partitioning strategy operating in FL478. A very low tissue Na+/K+ ratio in the leaves of FL478 after 7 days of stress hinted the existence of selective ion transport mechanism in this genotype. On the contrary, Kamini, an equally salt-tolerant genotype, was found to possess a higher leaf Na+/K+ ratio than does FL478 under similar stress condition. Salt-induced expression of different Na+ and K+ transporters indicated significant upregulation of SOS, HKT, NHX, and HAK groups of transporters in both leaves and roots of FL478, followed by Kamini. The expression of plasma membrane and vacuolar H+ pumps (OsAHA1, OsAHA7, and OsV-ATPase) were also upregulated in these two genotypes. On the other hand, IR29 and AC847 showed greater salt susceptibility owing to excess upward transport of Na+ and eventually died within a few days of stress imposition. But in the “leaf clip” assay, it was found that both IR29 and Kamini had high tissue-tolerance and chlorophyll-retention abilities. On the contrary, FL478, although having higher ionic-discrimination ability, showed the least degree of tissue tolerance as evident from the LC50 score (amount of Na+ required to reduce the initial chlorophyll content to half) of 336 mmol g–1 as against 459 and 424 mmol g–1 for IR29 and Kamini, respectively. Overall, the present study indicated that two components (ionic selectivity and tissue tolerance) of salt tolerance mechanism are distinct in rice. Unique genotypes like Kamini could effectively balance both of these strategies to achieve considerable salt tolerance, perhaps with lesser energy cost.

Expression of plasma membrane H+‐ATPase in cluster roots of white lupin under phosphorus deficiency

AbstractIt was previously shown that plasma membrane (PM) H+‐ATPases of active cluster roots are acclimated to phosphorus (P) deficiency. In the present study, we followed the question whether the qualitative acclimation of PM H+‐ATPase is based on isoform‐specific expression. Therefore, three different PM H+‐ATPase isoforms (LHA1–LHA3) were investigated at the transcriptional level. Our data indicate that LHA1 may play a key role in the qualitative acclimation of PM H+‐ATPase in cluster roots to P deficiency.

Mitigation of Salinity Stress in Plants by Arbuscular Mycorrhizal Symbiosis: Current Understanding and New Challenges.

Modern agriculture is facing twin challenge of ensuring global food security and executing it in a sustainable manner. However, the rapidly expanding salinity stress in cultivable areas poses a major peril to crop yield. Among various biotechnological techniques being used to reduce the negative effects of salinity, the use of arbuscular mycorrhizal fungi (AMF) is considered to be an efficient approach for bio-amelioration of salinity stress. AMF deploy an array of biochemical and physiological mechanisms that act in a concerted manner to provide more salinity tolerance to the host plant. Some of the well-known mechanisms include improved nutrient uptake and maintenance of ionic homeostasis, superior water use efficiency and osmoprotection, enhanced photosynthetic efficiency, preservation of cell ultrastructure, and reinforced antioxidant metabolism. Molecular studies in past one decade have further elucidated the processes involved in amelioration of salt stress in mycorrhizal plants. The participating AMF induce expression of genes involved in Na+ extrusion to the soil solution, K+ acquisition (by phloem loading and unloading) and release into the xylem, therefore maintaining favorable Na+:K+ ratio. Colonization by AMF differentially affects expression of plasma membrane and tonoplast aquaporins (PIPs and TIPs), which consequently improves water status of the plant. Formation of AM (arbuscular mycorrhiza) surges the capacity of plant to mend photosystem-II (PSII) and boosts quantum efficiency of PSII under salt stress conditions by mounting the transcript levels of chloroplast genes encoding antenna proteins involved in transfer of excitation energy. Furthermore, AM-induced interplay of phytohormones, including strigolactones, abscisic acid, gibberellic acid, salicylic acid, and jasmonic acid have also been associated with the salt tolerance mechanism. This review comprehensively covers major research advances on physiological, biochemical, and molecular mechanisms implicated in AM-induced salt stress tolerance in plants. The review identifies the challenges involved in the application of AM in alleviation of salt stress in plants in order to improve crop productivity.

Intracellular Mechanism of Rosuvastatin-Induced Decrease in Mature hERG Protein Expression on Membrane.

The hERG potassium channel (IKr) encoded by human ether-a-go-go-related gene plays an important role in cardiac repolarization. Decreased IKr may lead to long QT syndrome, which subsequently causes torsade de pointes and sudden cardiac death. Previous studies have shown that statins inhibit IKr and are more potent in inhibiting hERG currents when combined with other drugs. Since chemical structure of rosuvastatin is similar to that of several IKr blockers (ibutilide and E-4031), the present study aimed to reveal the mechanism that underlies rosuvastatin-induced hERG current reduction and to evaluate the possibility of cardiac toxicity. The results showed that rosuvastatin reduced hERG currents by accelerating the inactivation and prolonged action potential duration (APD) in hiPSC-CMs. Meanwhile, it was observed that rosuvastatin reduced the expression of the mature hERG. Transcription factor Sp1 was involved in hERG protein downregulation induced by rosuvastatin, and the result was verified by Sp1 siRNA and Sp1 agonist epicatechin. These results indicated that rosuvastatin could potentially inhibit transcription and reduce hERG mRNA expression. The interaction between hERG and heat shock protein was evaluated to study the mechanism of trafficking inhibition through co-immunoprecipitation. We found that rosuvastatin reduces the interaction of heat shock protein 70 (Hsp70) with the hERG protein, thereby affecting the folding of the hERG channel. Additionally, rosuvastatin significantly activates ATF6, which plays a key role in the activation of the unfolded protein response (UPR) pathway. Increased expression of the molecular chaperone calnexin and calreticulin, which are activated by ATF6 to help channel folding, further confirmed UPR activation. Meanwhile, the degradation of the hERG channel was mediated by lysosomes and proteasomes. In conclusion, Rosuvastatin reduced the expression of hERG plasma membrane by two pathways, the first is to disrupt the transport of immature hERG channels to the membrane, and the second is to increase the degradation of mature hERG channels. In addition, Rosuvastatin potently blocked hERG current, delayed cardiac repolarization, and thereby prolonged APDs and QTc intervals. Therefore, caution should be taken when rosuvastatin is used in the treatment of hyperlipidemia, especially when combined with drugs that can prolong the QT interval.

De novo transcriptomics analysis revealed a global reprogramming towards dehydration and hyposalinity in Bangia fuscopurpurea gametophytes (Rhodophyta)

Bangia fuscopurpurea is a farmed species in the ancient family Bangiales. It inhabits upper intertidal zones and suffers periodical desiccation and osmotic stress. The transcriptomic regulation under dehydration and hyposalinity was investigated. The differentially expressed genes (DEGs) accounted for 18.7% of the unigenes obtained by de novo transcriptome assembly (|log2fold-change| ≥ 1, FDR ≤ 0.001). Over 72% of the DEGs were downregulated under stress. The DEGs were predominantly enriched into the KEGG pathways “metabolic pathways,” “ribosome,” “biosynthesis of secondary metabolites,” “protein processing in endoplasmic reticulum,” and “oxidative phosphorylation.” The optimum photosynthetic efficiency (Fv/Fm) and photochemical quenching (qP) dropped significantly with 89% relative water loss and recovered rapidly after being rehydrated. Most DEGs regarding “photosynthesis” and “C3 carbon fixation” were upregulated in the dehydrated thalli, which may enable the thalli to gain photosynthetic recovery once being rehydrated. Fv/Fm and qP decreased significantly with 1 h of 90% freshwater treatment and then recovered to the control level 1 day later. With 6 h hyposaline treatment, expression of plasma membrane H+-ATPase genes was strongly and predominantly induced while the mRNA abundance of vacuolar, chloroplastic, and mitochondrial H+-ATPase genes decreased or showed no significant change. Some transporter, ion channel, and transmembrane protein genes together with the gene-encoding key enzymes involving in proline and heteroside metabolism were upregulated under hyposalinity. The results indicated that transmembrane exchange of ion and osmolytes was induced under hyposalinity to balance the osmotic fluctuation, which seemed to be triggered by plasma membrane H+-ATPases. These findings will facilitate elucidating the stress acclimation mechanism of B. fuscopurpurea.

Effect of Lanthanum on Plasma Membrane H+-ATPase in Rice (Oryza sativa) Under Acid Rain Stress

Global food security is threatened by damage to crop production by acid rain. To alleviate acid rain damage, we studied the effects of lanthanum (La3+) (0.06 and 0.12 mmol L−1) on plasma membrane H+-ATPase in rice seedlings under acid rain stress (pH 3.5 and 2.5). Relative growth rate, intracellular H+, ATP content, the activity, and gene expression of plasma membrane H+-ATPase were measured to validate the association of La3+ and plasma membrane H+-ATPase. We found that 0.06 mmol L−1 La3+ increased the plasma membrane H+-ATPase activity in rice treated with acid rain (pH 3.5) by increasing transcript levels of OSA 1,OSA 5, and OSA 7 genes, and thus, beneficial to pump excess H+ out of cells by supplying ATP energy. Thus, the decrease in relative growth rate was alleviated because of the application of 0.06 mmol L−1 La3+, showing an antagonistic interaction of acid rain and La3+ (0.06 mmol L−1). Contrarily, the application of 0.12 mmol L−1 La3+ aggravated the decrease in relative growth rate of rice under acid rain by decreasing the activity and expression of plasma membrane H+-ATPase, showing a synergistic interaction of acid rain and La3+ (0.12 mmol L−1). These results indicate that La3+ at the proper concentration can enhance the tolerance of rice seedlings to acid rain stress by increasing the activity of plasma membrane H+-ATPase, whereas La3+ at a higher concentration can aggravate the damage caused by acid rain.

Humic acid differentially improves nitrate kinetics under low- and high-affinity systems and alters the expression of plasma membrane H+-ATPases and nitrate transporters in rice

Humic acids (HAs) have a major effect on nutrient uptake, metabolism, growth and development in plants. Here, we evaluated the effect of HA pretreatment applied with a nutrient solution on the uptake kinetics of nitrate nitrogen (N-NO3−) and the metabolism of nitrogen (N) in rice under conditions of high and low NO3− supply. In addition, the kinetic parameters of NO3− uptake, N metabolites, and nitrate transporters (NRTs) and the plasma membrane (PM) H+-ATPase gene expression were examined. The plants were grown in a growth chamber with modified Hoagland and Arnon solution until 21 days after germination (DAG), and they were then transferred to a solution without N for 48 h and then to another solution without N and with and without the addition of HAs for another 48 h. After this period of N deprivation, the plants received new nutrient solutions containing 0.2 and 2.0 mM N-NO3−. Treatment of rice plants with HA promoted the induction of the genes OsNRT2.1-2.2/OsNAR2.1 and some isoforms PM H+-ATPase in roots. The application of HAs differentially modified the parameters of the uptake kinetics of NO3− under both concentrations. When grown with 0.2 mM NO3−, the plants pretreated with HA had lower Km and Cmin values as well as a higher Vmax/Km ratio. When grown with 2 mM NO3−, the plants pretreated with HA had a higher Vmax value, a greater root and shoot mass, and a lower root/shoot ratio. The N fractions were also altered by pretreatment with HA, and a greater accumulation of NO3− and N-amino was observed in the roots and shoots, respectively, of plants pretreated with HA. The results suggest that pretreatment with HA modifies root morphology and gene expression of PM H+-ATPases and NO3− transporters, resulting in a greater efficiency of NO3− acquisition by high- and low-affinity systems.

B-lymphocytes expressing an Ig specificity recognizing the pancreatic ß-cell autoantigen peripherin are potent contributors to type 1 diabetes development in NOD mice.

Although the autoimmune destruction of pancreatic β-cells underlying type 1 diabetes (T1D) development is ultimately mediated by T cells in NOD mice and also likely in humans, B cells play an additional key pathogenic role. It appears that the expression of plasma membrane–bound Ig molecules that efficiently capture β-cell antigens allows autoreactive B cells that bypass normal tolerance induction processes to be the subset of antigen-presenting cells most efficiently activating diabetogenic T cells. NOD mice transgenically expressing Ig molecules recognizing antigens that are (insulin) or are not (hen egg lysozyme [HEL]) expressed by β-cells have proven useful in dissecting the developmental basis of diabetogenic B cells. However, these transgenic Ig specificities were originally selected for their ability to recognize insulin or HEL as foreign, rather than autoantigens. Thus, we generated and characterized NOD mice transgenically expressing an Ig molecule representative of a large proportion of naturally occurring islet-infiltrating B cells in NOD mice recognizing the neuronal antigen peripherin. Transgenic peripherin-autoreactive B cells infiltrate NOD pancreatic islets, acquire an activated proliferative phenotype, and potently support accelerated T1D development. These results support the concept of neuronal autoimmunity as a pathogenic feature of T1D, and targeting such responses could ultimately provide an effective disease intervention approach.