Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.6.1.3 (ATPase)
65,361 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

In soybean (Glycine max L.), salicylic acid (SA) is converted primarily to SA 2-O-beta-d-glucose (SAG) in the cytoplasm and then accumulates exclusively in the vacuole. However, the mechanism involved in the vacuolar transport of SAG has not been investigated. The vacuolar transport of SAG was characterized by measuring the uptake of [(14)C]SAG into tonoplast vesicles isolated from etiolated soybean hypocotyls. The uptake of SAG was stimulated about six-fold when MgATP was included in the assay media. In contrast, the uptake of SA was only stimulated 1.25-fold by the addition of MgATP and was 2.2-fold less than the uptake of SAG providing an indication that the vacuolar uptake of SA is promoted by glucosylation. The ATP-dependent uptake of SAG was inhibited by increasing concentrations of vanadate (64% inhibition in the presence of 500 microM) but was not very sensitive to inhibition by bafilomycin A(1) (a specific inhibitor of vacuolar H(+)-ATPase; EC 3.6.1.3), and dissipation of the transtonoplast H(+)-electrochemical gradient. The SAG uptake exhibited Michaelis-Menten-type saturation kinetics with a K(m) value of 90 microM for SAG. SAG uptake was inhibited 60% by beta-estradiol 17-(beta-d-glucuronide), but glutathione conjugates and uncharged glucose conjugates were only slightly inhibitory. Based on the characteristics of SAG uptake into soybean tonoplast vesicles it is likely that this uptake occurs through an ATP-binding cassette transporter-type mechanism. However, this vacuolar uptake mechanism is not universal since the uptake of SAG by red beet (Beta vulgaris L) tonoplast vesicles appears to involve an H(+)-antiport mechanism.
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PMID:Uptake of salicylic acid 2-O-beta-D-glucose into soybean tonoplast vesicles by an ATP-binding cassette transporter-type mechanism. 1503 22

Regulation of ion homeostasis is fundamental to physiological activities in plants. Here, we report on the functional characterization of AcPMP3 [Aneurolepidium chinense (a monocotyledonous halophyte) plasma membrane protein 3] under salt stress. Expression of AcPMP3-1 and AcPMP3-2 genes was highly induced by various abiotic stresses, such as salt, cold and drought. Furthermore, abscisic acid, H(2)O(2) and salicylic acid also triggered expression of AcPMP3 genes. In the Deltanha1 Deltapmr2 Deltapmp3 yeast mutant, which lacks the major Na(+) efflux systems (Na(+)/H(+) antiporter and Na(+)-ATPase), its salt-sensitive phenotype was restored by expressing the AcPMP3-1 gene, and the transformants accumulated lesser amounts of Na(+) and K(+) than mutant cells under 50 mM NaCl and 500 mM KCl conditions, respectively. These results suggested that AcPMP3-1 plays a role as a regulator of both Na(+) and K(+) accumulation in the cells. In situ hybridization showed that the AcPMP3-1 transcript was localized in cells of the root cap and root epidermis, which strongly suggested that AcPMP3-1 is essential for regulating Na(+)/K(+) transportation between plant roots and the outer environment under salt stress.
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PMID:A stress-inducible plasma membrane protein 3 (AcPMP3) in a monocotyledonous halophyte, Aneurolepidium chinense, regulates cellular Na(+) and K(+) accumulation under salt stress. 1558 May 28

The hypersensitive response (HR) is one of the most critical defense systems in higher plants. In order to understand its molecular basis, we have screened tobacco genes that are transcriptionally activated during the early stage of the HR by the differential display method. Among six genes initially identified, one was found encoding a 57 kDa polypeptide with 497 amino acids not showing significant similarity to any reported proteins except for the AAA domain (ATPase associated with various cellular activities) spanning over 230 amino acids. The bacterially expressed protein exhibited ATP hydrolysis activity, and a green fluorescent protein-fusion protein localized in the cytoplasm of onion epidermis cells. The protein was subsequently designated as NtAAA1 (Nicotiana tabacum AAA1). NtAAA1 transcripts were induced 6 h after HR onset not only by TMV but also by incompatible Psuedomonas syringae, indicating that NtAAA1 is under the control of the N-gene with a common role in pathogen responses. Expression of NtAAA1 was induced by jasmonic acid and ethylene, but not by salicylic acid (SA). It also occurred at a high level in SA-deficient tobacco plants upon TMV infection. When NtAAA1 was silenced by the RNAi method, accumulation of transcripts for PR-1a significantly increased during the HR. Treatments with SA induced higher expression of PR-1a and acidic PR-2 in RNAi transgenic plants than in wild-type counterparts. These results suggest that NtAAA1 mitigates the SA signaling pathway, and therefore that NtAAA1 modulates the pathogen response of the host plants by adjusting the HR to an appropriate level.
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PMID:A hypersensitive response-induced ATPase associated with various cellular activities (AAA) protein from tobacco plants. 1582 94

The metabolism of salicylic acid (SA) in tobacco (Nicotiana tabacum L. cv. KY 14) cell suspension cultures was examined by adding [7-14C]SA to the cell cultures for 24 h and identifying the metabolites through high performance liquid chromatography analysis. The three major metabolites of SA were SA 2-O-beta-D: -glucose (SAG), methylsalicylate 2-O-beta-D: -glucose (MeSAG) and methylsalicylate. Studies on the intracellular localization of the metabolites revealed that all of the SAG associated with tobacco protoplasts was localized in the vacuole. However, the majority of the MeSAG was located outside the vacuole. The tobacco cells contained an SA inducible SA glucosyltransferase (SAGT) enzyme that formed SAG. The SAGT enzyme was not associated with the vacuole and appeared to be a cytoplasmic enzyme. The vacuolar transport of SAG was characterized by measuring the uptake of [14C]SAG into tonoplast vesicles isolated from tobacco cell cultures. SAG uptake was stimulated eightfold by the addition of MgATP. The ATP-dependent uptake of SAG was inhibited by bafilomycin A1 (a specific inhibitor of the vacuolar H(+)-ATPase) and dissipation of the transtonoplast H(+)-electrochemical gradient. Vanadate was not an inhibitor of SAG uptake. Several beta-glucose conjugates were strong inhibitors of SAG uptake, whereas glutathione and glucuronide conjugates were only marginally inhibitory. The SAG uptake exhibited Michaelis-Menten type saturation kinetics with a K(m) and V(max) value of 11 microM and 205 pmol min-1 mg-1, respectively, for SAG. Based on the transport characteristics it appears as if the vacuolar uptake of SAG in tobacco cells occurs through an H(+)-antiport-type mechanism.
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PMID:The formation, vacuolar localization, and tonoplast transport of salicylic acid glucose conjugates in tobacco cell suspension cultures. 1587 Oct 31

The hypersensitive response (HR) is defined as rapid cell collapse at the infection site and often accompanies plant resistance. The physiological processes leading to HR are not well understood. Here, we report an electrophysiological characterization of bacterial HR caused by a single avirulence gene in the absence of other bacterial signals. We used dexamethasone (dex)-inducible transgenic Arabidopsis (Arabidopsis thaliana) plants containing the avrRpt2 gene from Pseudomonas syringae pv tomato. Membrane depolarization in these plants began 1 to 1.5 h after dex application, hours before electrolyte leakage. Progressive depolarization was a sensitive early indicator of HR that occurred only in Arabidopsis leaf cells expressing both avrRpt2 and a functional RPS2 gene. Hyperpolarization of fully depolarized membranes by fusicoccin, a fungal toxin that activates the H(+)-ATPase, indicates that depolarization did not result from a nonfunctional pump or leaky membranes. Depolarization and electrolyte leakage were inhibited in RPS2 plants by the calcium channel blocker LaCl(3), highly correlating these events and suggesting that Ca(2+) entry into cells is required for both. Also correlated were inhibition of depolarization, electrolyte leakage, and HR following salicylic acid pretreatment. In salicylic acid-pretreated RPS2 seedlings, avrRpt2 transcript was produced after dex treatment. However, AvrRpt2 protein accumulation was greatly reduced, suggesting a possible mechanism for inhibition of HR in plants with induced resistance. This experimental system is a very sensitive assay that lends itself to the dissection of physiological processes leading to HR in plants, and provides a baseline for future research within a genetic framework.
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PMID:Electrophysiological characterization of the Arabidopsis avrRpt2-specific hypersensitive response in the absence of other bacterial signals. 1590 9

We have cloned and characterized the cDNA, genomic clone and upstream promoter region of a vacuolar ATPase (V-ATPase) c subunit (PgVHA-c1) from Pennisetum glaucum. The deduced amino acid sequence shows 98-71% sequence identity with V-ATPase from rice and Arabidopsis, and is a highly hydrophobic protein with four transmembrane regions. PgVHA-c1-GFP fusion protein is expressed in BY2 cells on the endo-membranes surrounding vacuoles; however, PgVHA-c1 could not functionally complement V-ATPase-c deletion mutants of yeast. The sequence analysis of the genomic clone revealed the presence of two introns in the coding region, and the splice junctions followed the typical canonical GU-AG consensus sequence. The transcript analysis showed that the expression of PgVHA-c1 was stimulated more in response to salinity stress and very marginally in response to drought and low temperature stress. Exogenous application of abscisic acid, salicylic acid and calcium stimulated the transcript level in the absence of stress. We have cloned the 5'-flanking regions of PgVHA-c1 and mapped its transcript start site at 78 bp upstream of ATG. Transgenic tobacco with promoter::GUS constructs showed that the region -288/+78 was sufficient for GUS expression. The expression of the reporter gene even with the full-length promoter was limited to shoot hairs and to male and female reproductive organs. The dehydration-responsive element (DRE) and ABA-responsive element (ABRE) in the promoter did not show consensus flanking regions; however, gel mobility shift assays showed that Pennisetum has specific transacting factors that showed binding to the core DRE, ABRE and TCA elements.
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PMID:Cloning and regulation of a stress-regulated Pennisetum glaucum vacuolar ATPase c gene and characterization of its promoter that is expressed in shoot hairs and floral organs. 1595 96

Fusarium spp. are ubiquitous fungi found in soil worldwide as both pathogenic and nonpathogenic strains. The signals leading to disease or the absence of disease are poorly understood. We recently showed that fusaric acid (FA), a nonspecific toxin produced by most Fusarium spp., could elicit various plant defense responses at 100 nM without toxic effect. In this study, we checked for the effect of FA on root and root hairs, probable first site of contact between the fungi and the host. Large FA concentrations reduce root and root-hair growth and induce a rapid transient membrane hyperpolarization, followed by a large depolarization, due to the inhibition of H(+)-ATPase currents. Nanomolar concentrations of FA induced only an early transient membrane hyperpolarization of root hairs compatible with the induction of a signal transduction pathway. FA at 10(-7) M failed to induce salicylic acid- and jasmonic acid/ethylene-dependent defense-related genes but inhibited the germination of the angiosperm parasite Orobanche ramosa in contact of FA-pretreated Arabidopsis thaliana seedlings. These data suggest that FA at nontoxic concentrations could activate signal transduction components necessary for plant-defense responses that could contribute to biocontrol activity of Fusarium spp.
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PMID:A putative role for fusaric acid in biocontrol of the parasitic angiosperm Orobanche ramosa. 1667 42

The aim of this study was to evaluate the effect of 2,5-dihydroxybenzoic acid, a salicylate derived from Acetyl salicylic acid (ASA) and vitamin A (vit A) on Na(+), K(+) ATPase enzyme and GSH levels in brain of rats exposed to hyperoxia (Hyp) as oxidant protocol. Rats were treated as follow: group I (control), group II (Hyp), group III (Hyp, ASA), group IV (vit A), group V (Hyp, vit A), group VI (Hyp, vit A, ASA). Vit A was given 5 days before and during Hyp, aspirin at the end of Hyp. Na(+),K(+) ATPase and total ATPase activity was significantly increased in group V. Levels of GSH showed a significant increase in group III, besides, levels of 2,5-dihydroxybenzoic acid as salicylate in plasma were significantly increased in group II. These results elucidate differences in the biochemical response of animal towards intake of various types of antioxidant substances, with increased GSH and salicylate in hyperoxia.
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PMID:Assessment of antioxidant effect of 2,5-dihydroxybenzoic acid and vitamin a in brains of rats with induced hyperoxia. 1740 73

ATPase associated with various cellular activities (AAA) proteins are commonly distributed among eukaryotes, and are involved in a multitude of cellular functions. NtAAA1 is one such example, being involved in pathogen response in tobacco plants. When its activity was suppressed in RNAi transgenic tobacco plants, an elevated resistance to the pathogenic bacterium Pseudomonas syringae was observed in comparison with the wild type. As AAA proteins function through interaction with specific partners, NtAAA1-interacting proteins were screened by the yeast two-hybrid assay, and one particular gene encoding a small GTPase, an ADP ribosylation factor, was identified and designated as NtARF. Its specific binding to NtAAA1 was confirmed by in vitro pull-down assay, and their interaction was predominant between active forms of NtARF and NtAAA1, each bound to GTP and ATP, respectively. Their physical interaction in vivo around the plasma membrane was shown by fluorescence resonance energy transfer assays, suggesting their role in membrane trafficking. Transgenic tobacco plants constitutively expressing NtARF under the control of a cauliflower mosaic virus 35S promoter exhibited spontaneous and wound-induced lesion formation, and enhanced resistance to pathogen attack. Expression of NtAAA1 in leaves of NtARF transgenic plants attenuated lesion and suppressed pathogen resistance. In wild-type tobacco plants, transcripts of NtAAA1 and NtARF could be induced by ethylene and salicylic acid, respectively. These results suggest that NtAAA1 balances plant resistance through suppression of NtARF, and that the molecular basis for the known antagonistic actions of ethylene and salicylic acid in defense response could be partly attributable to these two proteins.
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PMID:Attenuation of the hypersensitive response by an ATPase associated with various cellular activities (AAA) protein through suppression of a small GTPase, ADP ribosylation factor, in tobacco plants. 1755 12

H(2)O(2), plasma membrane H(+)-ATPase (PM H(+)-ATPase) and salicylic acid (SA) play important roles in sensing external stimulation and activating defense responses in plants. However, it remains uncertain whether they are involved and interrelated in response to heat acclimation. Experiments were performed by pharmacological methods, and the relationship and the connection between endogenous H(2)O(2), free SA and PM H(+)-ATPase were investigated in pea plants (Pisum sativum L.) during heat acclimation. The results showed that an accumulation peaks of H(2)O(2), free SA and PM H(+)-ATPase, were detected during heat acclimation at 37 degrees C for 2 h and H(2)O(2) burst appeared before SA accumulation that followed by increase of PM H(+)-ATPase activity (Fig.1). Pretreatments with either scavengers of active oxygen species (dimethyl sulfoxide and ascorbic acid) or antioxidant (reduced glutathione) inhibited the increases in both H(2)O(2) and free SA contents as a part of heat acclimation (Fig.2). Additionally, changes in activity of plasma membrane NADPH oxidase paralleled with H(2)O(2) level during heat acclimation (Figs.1 and 3), implicating that H(2)O(2) might be generated by plasma membrane NADPH oxidase. Moreover, pretreatments with either diphenylene iodonium (DPI), a suicide substrate inhibitor of plasma membrane NADPH oxidase, or dimethylthiourea (DMTU), a quencher of H(2)O(2), could block the increase in free SA content and activity of plasma membrane NADPH oxidase as a part of heat acclimation (Fig.4). According to the assay described above, it is suggested that both H(2)O(2) and PM H(+)-ATPase participate in SA signaling that leads to the development of thermotolerance in pea plant, and H(2)O(2) functions upstream and PM H(+)-ATPase functions downstream of the SA signal. Also, the regulation mechanism of PM H(+)-ATPase activity was investigated, which showed that during heat acclimation, increase of PM H(+)-ATPase activity was independent of PM H(+)-ATPase amount and the enzyme activity may be modulated at post-translational level that may involve in reversible protein phosphorylation (Fig.5).
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PMID:[Changes in H2O2 and salicylic acid contents as well as plasma membrane H+-ATPase activity and their relations in pea leaves during thermotolerance induction]. 1796 46


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