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1.
Biomolecules ; 14(7)2024 Jul 05.
Article de Anglais | MEDLINE | ID: mdl-39062513

RÉSUMÉ

Lowe Syndrome (LS) is a rare X-linked disorder characterized by renal dysfunction, cataracts, and several central nervous system (CNS) anomalies. The mechanisms underlying the neurological dysfunction in LS remain unclear, albeit they share some phenotypic characteristics similar to the deficiency or dysfunction of the Reelin signaling, a relevant pathway with roles in CNS development and neuronal functions. In this study, we investigated the role of OCRL1, an inositol polyphosphate 5-phosphatase encoded by the OCRL gene, mutated in LS, focusing on its impact on endosomal trafficking and receptor recycling in human neuronal cells. Specifically, we tested the effects of OCRL1 deficiency in the trafficking and signaling of ApoER2/LRP8, a receptor for the ligand Reelin. We found that loss of OCRL1 impairs ApoER2 intracellular trafficking, leading to reduced receptor expression and decreased levels at the plasma membrane. Additionally, human neurons deficient in OCRL1 showed impairments in ApoER2/Reelin-induced responses. Our findings highlight the critical role of OCRL1 in regulating ApoER2 endosomal recycling and its impact on the ApoER2/Reelin signaling pathway, providing insights into potential mechanisms underlying the neurological manifestations of LS.


Sujet(s)
Molécules d'adhérence cellulaire neuronale , Endosomes , Protéines de la matrice extracellulaire , Protéines apparentées au récepteur LDL , Protéines de tissu nerveux , Neurones , Phosphoric monoester hydrolases , Transport des protéines , Protéine reeline , Serine endopeptidases , Humains , Phosphoric monoester hydrolases/métabolisme , Phosphoric monoester hydrolases/génétique , Phosphoric monoester hydrolases/déficit , Protéines de tissu nerveux/métabolisme , Protéines de tissu nerveux/génétique , Protéines de tissu nerveux/déficit , Serine endopeptidases/métabolisme , Serine endopeptidases/génétique , Serine endopeptidases/déficit , Molécules d'adhérence cellulaire neuronale/métabolisme , Molécules d'adhérence cellulaire neuronale/génétique , Molécules d'adhérence cellulaire neuronale/déficit , Protéines de la matrice extracellulaire/métabolisme , Protéines de la matrice extracellulaire/génétique , Protéines de la matrice extracellulaire/déficit , Endosomes/métabolisme , Neurones/métabolisme , Protéines apparentées au récepteur LDL/métabolisme , Protéines apparentées au récepteur LDL/génétique , Transduction du signal , Syndrome de Lowe/génétique , Syndrome de Lowe/métabolisme
2.
New Phytol ; 243(1): 132-144, 2024 Jul.
Article de Anglais | MEDLINE | ID: mdl-38742309

RÉSUMÉ

Nutrient limitation may constrain the ability of recovering and mature tropical forests to serve as a carbon sink. However, it is unclear to what extent trees can utilize nutrient acquisition strategies - especially root phosphatase enzymes and mycorrhizal symbioses - to overcome low nutrient availability across secondary succession. Using a large-scale, full factorial nitrogen and phosphorus fertilization experiment of 76 plots along a secondary successional gradient in lowland wet tropical forests of Panama, we tested the extent to which root phosphatase enzyme activity and mycorrhizal colonization are flexible, and if investment shifts over succession, reflective of changing nutrient limitation. We also conducted a meta-analysis to test how tropical trees adjust these strategies in response to nutrient additions and across succession. We find that tropical trees are dynamic, adjusting investment in strategies - particularly root phosphatase - in response to changing nutrient conditions through succession. These changes reflect a shift from strong nitrogen to weak phosphorus limitation over succession. Our meta-analysis findings were consistent with our field study; we found more predictable responses of root phosphatase than mycorrhizal colonization to nutrient availability. Our findings suggest that nutrient acquisition strategies respond to nutrient availability and demand in tropical forests, likely critical for alleviating nutrient limitation.


Sujet(s)
Forêts , Mycorhizes , Azote , Nutriments , Phosphore , Arbres , Climat tropical , Phosphore/métabolisme , Azote/métabolisme , Mycorhizes/physiologie , Nutriments/métabolisme , Racines de plante/métabolisme , Racines de plante/microbiologie , Phosphoric monoester hydrolases/métabolisme , Panama
3.
Mol Microbiol ; 120(1): 20-31, 2023 07.
Article de Anglais | MEDLINE | ID: mdl-37042030

RÉSUMÉ

In the last two decades, an increasing number of bacterial species have been recognized that are able to generate a phenotypically diverse population that shares an identical genotype. This ability is dependent on a complex genetic regulatory network that includes cellular and environmental signals, as well as stochastic elements. Among Bacilli, a broadly distributed family of Rap (Response-regulator aspartyl phosphate) phosphatases is known to modulate the function of the main phenotypic heterogeneity regulators by controlling their phosphorylation. Even more, their related extracellular Phr (Phosphatase regulator) peptides function as signals, creating a cell-cell communication network that regulates the phenotypic development of the entire population. In this review, we examine the role that the Rap phosphatases and their Phr peptides play in the regulation of Bacillus subtilis phenotypic differentiation, and in other members of the Bacillus genus. We also highlight the contribution of these regulatory elements to the fitness of bacterial cells and mobile genetic elements, for example, prophages and conjugative vectors.


Sujet(s)
Bacillus , Phosphoric monoester hydrolases , Phosphoric monoester hydrolases/génétique , Phosphoric monoester hydrolases/métabolisme , Bacillus/génétique , Réseaux de régulation génique , Protéines bactériennes/génétique , Protéines bactériennes/métabolisme , Peptides/génétique , Bacillus subtilis/métabolisme , Adaptation physiologique , Régulation de l'expression des gènes bactériens/génétique
4.
Methods Mol Biol ; 2555: 103-114, 2023.
Article de Anglais | MEDLINE | ID: mdl-36306081

RÉSUMÉ

Phosphate release from inorganic and organic phosphorus compounds can be enzymatically mediated. Phosphate-releasing enzymes, comprising acid and alkaline phosphatases, are recognized as useful biocatalysts in applications such as plant and animal nutrition, bioremediation, and diagnostic analysis. Here, we describe a functional metagenomics approach enabling rapid identification of genes encoding these enzymes. The target genes are detected based on small- and large-insert metagenomic libraries derived from diverse environments. This approach has the potential to unveil entirely new phosphatase families or subfamilies and members of known enzyme classes that hydrolyze phosphomonoester bonds such as phytases. Additionally, we provide a strategy for efficient heterologous expression of phosphatase genes.


Sujet(s)
Phytase , Métagénomique , Métagénome , Phosphoric monoester hydrolases/génétique , Phosphoric monoester hydrolases/métabolisme , Phytase/génétique , Phosphates
5.
Int J Mol Sci ; 23(23)2022 Dec 01.
Article de Anglais | MEDLINE | ID: mdl-36499456

RÉSUMÉ

Matrix vesicles (MVs) contain the whole machinery necessary to initiate apatite formation in their lumen. We suspected that, in addition to tissue-nonspecific alkaline phosphatase (TNAP), Na,K,-ATPase (NKA) could be involved in supplying phopshate (Pi) in the early stages of MV-mediated mineralization. MVs were extracted from the growth plate cartilage of chicken embryos. Their average mean diameters were determined by Dynamic Light Scattering (DLS) (212 ± 19 nm) and by Atomic Force Microcopy (AFM) (180 ± 85 nm). The MVs had a specific activity for TNAP of 9.2 ± 4.6 U·mg-1 confirming that the MVs were mineralization competent. The ability to hydrolyze ATP was assayed by a colorimetric method and by 31P NMR with and without Levamisole and SBI-425 (two TNAP inhibitors), ouabain (an NKA inhibitor), and ARL-67156 (an NTPDase1, NTPDase3 and Ecto-nucleotide pyrophosphatase/phosphodiesterase 1 (NPP1) competitive inhibitor). The mineralization profile served to monitor the formation of precipitated calcium phosphate complexes, while IR spectroscopy allowed the identification of apatite. Proteoliposomes containing NKA with either dipalmitoylphosphatidylcholine (DPPC) or a mixture of 1:1 of DPPC and dipalmitoylphosphatidylethanolamine (DPPE) served to verify if the proteoliposomes were able to initiate mineral formation. Around 69-72% of the total ATP hydrolysis by MVs was inhibited by 5 mM Levamisole, which indicated that TNAP was the main enzyme hydrolyzing ATP. The addition of 0.1 mM of ARL-67156 inhibited 8-13.7% of the total ATP hydrolysis in MVs, suggesting that NTPDase1, NTPDase3, and/or NPP1 could also participate in ATP hydrolysis. Ouabain (3 mM) inhibited 3-8% of the total ATP hydrolysis by MVs, suggesting that NKA contributed only a small percentage of the total ATP hydrolysis. MVs induced mineralization via ATP hydrolysis that was significantly inhibited by Levamisole and also by cleaving TNAP from MVs, confirming that TNAP is the main enzyme hydrolyzing this substrate, while the addition of either ARL-6715 or ouabain had a lesser effect on mineralization. DPPC:DPPE (1:1)-NKA liposome in the presence of a nucleator (PS-CPLX) was more efficient in mineralizing compared with a DPPC-NKA liposome due to a better orientation of the NKA active site. Both types of proteoliposomes were able to induce apatite formation, as evidenced by the presence of the 1040 cm-1 band. Taken together, the findings indicated that the hydrolysis of ATP was dominated by TNAP and other phosphatases present in MVs, while only 3-8% of the total hydrolysis of ATP could be attributed to NKA. It was hypothesized that the loss of Na/K asymmetry in MVs could be caused by a complete depletion of ATP inside MVs, impairing the maintenance of symmetry by NKA. Our study carried out on NKA-liposomes confirmed that NKA could contribute to mineral formation inside MVs, which might complement the known action of PHOSPHO1 in the MV lumen.


Sujet(s)
Calcinose , Phosphoric monoester hydrolases , Animaux , Embryon de poulet , Phosphoric monoester hydrolases/métabolisme , Sodium-Potassium-Exchanging ATPase , Calcification physiologique , Phosphatase alcaline/métabolisme , Hydrolyse , Adénosine triphosphate , Liposomes/composition chimique , Minéraux/métabolisme
6.
Life Sci ; 308: 120962, 2022 Nov 01.
Article de Anglais | MEDLINE | ID: mdl-36113732

RÉSUMÉ

l-Amino acid oxidase isolated from Calloselasma rhodostoma (Cr-LAAO) snake venom is a potent stimulus for neutrophil activation and production of inflammatory mediators, contributing to local inflammatory effects in victims of envenoming. Cr-LAAO triggered the activation of nicotinamide adenine dinucleotide phosphatase (NADPH) oxidase complex and protein kinase C (PKC)-α signaling protein for reactive oxygen species (ROS) production. This study aims to evaluate the ROS participation in the NLRP3 inflammasome complex activation in human neutrophil. Human neutrophils were isolated and stimulated for 1 or 2 h with RPMI (negative control), LPS (1 µg/mL, positive control) or Cr-LAAO (50 µg/mL). The neutrophil transcriptome was examined using the microarray technique, and RT-qPCR for confirmation of gene expression. Immunofluorescence assays for NLRP3, caspase-1, IL-1ß and GSDMD proteins was performed by Western blot in the presence and/or absence of Apocynin, an inhibitor of NADPH oxidase. IL-1ß release was also detected in the presence and/or absence of NLRP3, caspase-1 and NADPH oxidase inhibitors. Results showed that Cr-LAAO upregulated the expression of genes that participate in the NADPH oxidase complex formation and inflammasome assembly. NLRP3 was activated and accumulated in the cytosol forming punctas, indicating its activation. Gasdermin D was not cleaved but lactate dehydrogenase was released. Furthermore, ROS inhibition decreased the expression of NLRP3 inflammasome complex proteins, as observed by protein expression in the presence and/or absence of apocynin, an NADPH oxidase inhibitor. IL-1ß was also released, and pharmacological inhibition of NLRP3, caspase-1, and ROS reduced the amount of released cytokine. This is the first report demonstrating the activation of the NLRP3 inflammasome complex via ROS generation by Cr-LAAO, which may lead to the development of local inflammatory effects observed in snakebite victims.


Sujet(s)
Inflammasomes , L-Amino acid oxidase , Acétophénones , Caspase-1/métabolisme , Cytokines/métabolisme , Humains , Inflammasomes/métabolisme , Médiateurs de l'inflammation/métabolisme , Interleukine-1 bêta/métabolisme , L-Amino acid oxidase/métabolisme , L-Amino acid oxidase/pharmacologie , Lactate dehydrogenases/métabolisme , Lipopolysaccharides/pharmacologie , NAD/métabolisme , NADP/métabolisme , NADPH oxidase/métabolisme , Protéine-3 de la famille des NLR contenant un domaine pyrine/génétique , Protéine-3 de la famille des NLR contenant un domaine pyrine/métabolisme , Granulocytes neutrophiles/métabolisme , Phosphoric monoester hydrolases/métabolisme , Protéine kinase C/métabolisme , Espèces réactives de l'oxygène/métabolisme , Venins de serpent/métabolisme , Venins de serpent/pharmacologie
7.
Clin Immunol ; 236: 108952, 2022 03.
Article de Anglais | MEDLINE | ID: mdl-35149196

RÉSUMÉ

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease associated with multiple phenotypic and functional aberrations in T lymphocytes. Among these, altered expression and/or activity of several protein kinases and phosphatases has been consistently documented in T cells obtained from patients with SLE. In this review, we describe and contextualize some of the kinase and phosphatase defects reported in T cells from patients with SLE, highlighting their relevance and possible consequences. Additionally, we discuss the origin of the defects and its significance for disease development and expression.


Sujet(s)
Maladies auto-immunes , Lupus érythémateux disséminé , Maladies auto-immunes/métabolisme , Humains , Phosphoric monoester hydrolases/métabolisme , Protein kinases/métabolisme , Lymphocytes T
8.
Int J Mol Sci ; 22(4)2021 Feb 22.
Article de Anglais | MEDLINE | ID: mdl-33671604

RÉSUMÉ

Phosphatases are hydrolytic enzymes that cleave the phosphoester bond of numerous substrates containing phosphorylated residues. The typical classification divides them into acid or alkaline depending on the pH at which they have optimal activity. The histidine phosphatase (HP) superfamily is a large group of functionally diverse enzymes characterized by having an active-site His residue that becomes phosphorylated during catalysis. HP enzymes are relevant biomolecules due to their current and potential application in medicine and biotechnology. Entamoeba histolytica, the causative agent of human amoebiasis, contains a gene (EHI_146950) that encodes a putative secretory acid phosphatase (EhHAPp49), exhibiting sequence similarity to histidine acid phosphatase (HAP)/phytase enzymes, i.e., branch-2 of HP superfamily. To assess whether it has the potential as a biocatalyst in removing phosphate groups from natural substrates, we studied the EhHAPp49 structural and functional features using a computational-experimental approach. Although the combined outcome of computational analyses confirmed its structural similarity with HP branch-2 proteins, the experimental results showed that the recombinant enzyme (rEhHAPp49) has negligible HAP/phytase activity. Nonetheless, results from supplementary activity evaluations revealed that rEhHAPp49 exhibits Mg2+-dependent alkaline pyrophosphatase activity. To our knowledge, this study represents the first computational-experimental characterization of EhHAPp49, which offers further insights into the structure-function relationship and the basis for future research.


Sujet(s)
Entamoeba histolytica/enzymologie , Phosphoric monoester hydrolases/composition chimique , Phosphoric monoester hydrolases/métabolisme , Relation structure-activité , Phytase/métabolisme , Sites de fixation , Domaine catalytique , Diphosphates/métabolisme , Entamoeba histolytica/génétique , Humains , Simulation de docking moléculaire , Phosphoric monoester hydrolases/génétique , Conformation des protéines , Protéines de protozoaire/composition chimique , Protéines de protozoaire/génétique , Protéines de protozoaire/métabolisme , Protéines recombinantes/génétique , Protéines recombinantes/métabolisme
9.
J Inorg Biochem ; 219: 111392, 2021 06.
Article de Anglais | MEDLINE | ID: mdl-33752123

RÉSUMÉ

Herein, we report the synthesis and characterization of the first two AlIII(µ-OH)MII (M = Zn (1) and Cu (2)) complexes with the unsymmetrical ligand H2L{2-[[(2-hydroxybenzyl)(2-pyridylmethyl)]aminomethyl]-6-bis(pyridylmethyl)aminomethyl}-4-methylphenol. The complexes were characterized through elemental analysis, X-ray crystallography, IR spectroscopy, mass spectrometry and potentiometric titration. In addition, complex 2 was characterized by electronic spectroscopy. Kinetics studies on the hydrolysis of the model substrate bis(2,4-dinitrophenyl)phosphate by 1 and 2 show Michaelis-Menten behavior, with 1 being slightly more active (8.31%) than 2 (at pH 7.0). The antimicrobial effect of the compounds was studied using four bacterial strains (Staphylococcus aureus, Pseudomonas aeuruginosa, Shigella sonnei and Shigella dysenteriae) and for both complexes the inhibition of bacterial growth was superior to that caused by sulfapyridine, but inferior to that of tetracycline. The dark cytotoxicity and photocytotoxicity (under UV-A light) of the complexes in a chronic myelogenous leukemia cell line were investigated. Complexes 1 and 2 exhibited significant cytotoxic activity against K562 cells, which undergoes a 2-fold increase on applying 5 min of irradiation with UV-A light. Complex 2 was more effective and a good correlation between cytotoxicity and intracellular concentration was observed, the intracellular copper concentration required to inhibit 50% of cell growth being 3.5 × 10-15 mol cell-1.


Sujet(s)
Aluminium/pharmacologie , Antinéoplasiques/pharmacologie , Complexes de coordination/pharmacologie , Cuivre/pharmacologie , Phosphoric monoester hydrolases/métabolisme , Zinc/pharmacologie , Aluminium/composition chimique , Anti-infectieux/pharmacologie , Bactéries/effets des médicaments et des substances chimiques , Survie cellulaire/effets des médicaments et des substances chimiques , Complexes de coordination/composition chimique , Cuivre/composition chimique , Cristallographie aux rayons X/méthodes , Humains , Hydrolyse , Cellules K562 , Cinétique , Ligands , Spectrométrie de masse/méthodes , Zinc/composition chimique
10.
Plant Sci ; 304: 110808, 2021 Mar.
Article de Anglais | MEDLINE | ID: mdl-33568304

RÉSUMÉ

Iron (Fe) is an essential micronutrient for plants and is present abundantly in the Earth's crust. However, Fe bioavailability in alkaline soils is low due to the decreased solubility of the ferric ions. Previously, we have demonstrated the relationship between the PAP/SAL1 retrograde signaling pathway, the activity of Strategy I Fe uptake genes (FIT, FRO2, IRT1), and ethylene signaling. In this work, we have characterized mutant lines that are deficient in this retrograde signaling pathway and their ability to grow in alkaline soils. This adverse growth condition caused less impact on mutant plants, which showed less reduced rosette area, and higher carotenoid, chlorophyll and Fe content than wild-type plants. Several genes involved in the biosynthesis and excretion of secondary metabolites derived from the phenylpropanoid pathway, which improve Fe uptake, were elevated in mutant plants. Finally, we observed an increase in excreted fluorescent phenolic compounds in mutant lines compared to wild-type plants. In this way, PAP/SAL1 mutants showed alterations in the biosynthesis of metabolites that mobilize Fe, which ultimately improved these plants ability to grow in alkaline soils. Results agree with the existence of a link between the PAP/SAL1 retrograde signaling pathway and the regulation of Fe deficiency responses in Arabidopsis.


Sujet(s)
Protéines d'Arabidopsis/métabolisme , Arabidopsis/métabolisme , Carences en fer , PAPS/métabolisme , Phosphoric monoester hydrolases/métabolisme , Transduction du signal , Arabidopsis/physiologie , Concentration en ions d'hydrogène , Fer/métabolisme , Réaction de polymérisation en chaine en temps réel , Sol/composition chimique
11.
Biochim Biophys Acta Mol Cell Res ; 1868(3): 118927, 2021 03.
Article de Anglais | MEDLINE | ID: mdl-33310067

RÉSUMÉ

Platelets are small enucleated cell fragments specialized in the control of hemostasis, but also playing a role in angiogenesis, inflammation and immunity. This plasticity demands a broad range of physiological processes. Platelet functions are mediated through a variety of receptors, the concerted action of which must be tightly regulated, in order to allow specific and timely responses to different stimuli. Protein phosphorylation is one of the main key regulatory mechanisms by which extracellular signals are conveyed. Despite the importance of platelets in health and disease, the molecular pathways underlying the activation of these cells are still under investigation. Here, we review current literature on signaling platelet biology and in particular emphasize the newly emerging role of phosphatases in these processes.


Sujet(s)
Plaquettes/métabolisme , Phosphoric monoester hydrolases/métabolisme , Tyrosine/métabolisme , Régulation de l'expression des gènes , Hémostase , Humains , Phosphorylation , Transduction du signal
12.
Int J Mol Sci ; 21(16)2020 Aug 13.
Article de Anglais | MEDLINE | ID: mdl-32823483

RÉSUMÉ

Intestinal ischemia reperfusion injury (iIRI) is a severe clinical condition presenting high morbidity and mortality worldwide. Some of the systemic consequences of IRI can be prevented by applying ischemic preconditioning (IPC), a series of short ischemia/reperfusion events preceding the major ischemia. Although neutrophils are key players in the pathophysiology of ischemic injuries, neither the dysregulation presented by these cells in iIRI nor the protective effect of iIPC have their regulation mechanisms fully understood. Protein phosphorylation, as well as the regulation of the respective phosphatases and kinases are responsible for regulating a large number of cellular functions in the inflammatory response. Moreover, in previous work we found hydrolases and transferases to be modulated in iIR and iIPC, suggesting the possible involvement of phosphatases and kinases in the process. Therefore, in the present study, we analyzed the phosphoproteome of neutrophils from rats submitted to mesenteric ischemia and reperfusion, either submitted or not to IPC, compared to quiescent controls and sham laparotomy. Proteomic analysis was performed by multi-step enrichment of phosphopeptides, isobaric labeling, and LC-MS/MS analysis. Bioinformatics was used to determine phosphosite and phosphopeptide abundance and clustering, as well as kinases and phosphatases sites and domains. We found that most of the phosphorylation-regulated proteins are involved in apoptosis and migration, and most of the regulatory kinases belong to CAMK and CMGC families. An interesting finding revealed groups of proteins that are modulated by iIR, but such modulation can be prevented by iIPC. Among the regulated proteins related to the iIPC protective effect, Vamp8 and Inpp5d/Ship are discussed as possible candidates for control of the iIR damage.


Sujet(s)
Intestins/anatomopathologie , Préconditionnement ischémique , Granulocytes neutrophiles/métabolisme , Phosphoprotéines/métabolisme , Phosphoric monoester hydrolases/métabolisme , Protein kinases/métabolisme , Protéomique , Lésion d'ischémie-reperfusion/métabolisme , Motifs d'acides aminés , Séquence d'acides aminés , Animaux , Phosphopeptides/composition chimique , Phosphopeptides/métabolisme , Phosphoprotéines/composition chimique , Phosphorylation , Domaines protéiques , Protéome/métabolisme , Rats , Lésion d'ischémie-reperfusion/anatomopathologie , Transduction du signal
13.
Sci Rep ; 10(1): 11365, 2020 07 09.
Article de Anglais | MEDLINE | ID: mdl-32647221

RÉSUMÉ

Worldwide, alien plant invasions have been intensively studied in the past decades, but mechanisms controlling the invasibility of native communities are not fully understood yet. The stochastic niche hypothesis predicts that species-rich plant communities are less prone to alien plant invasions than species-poor communities, which is supported by some but not all field studies, with some very species-rich communities such as the Brazilian Cerrado becoming heavily invaded. However, species-rich communities potentially contain a greater variety of facilitative interactions in resource exploitation than species-poor communities, from which invasive plants might benefit. This alternative hypothetical mechanism might explain why nutrient-poor, species-rich ecosystems are prone to invasion. Here we show that a high species richness both impedes and promotes invasive plants in the Brazilian Cerrado, using structural equation modelling and data from 38 field sites. We found support for the stochastic niche hypothesis through an observed direct negative influence of species richness on abundance of alien invasive species, but an indirect positive effect of species richness on invasive alien plants through soil phosphatase activity that enhances P availability was also found. These field observations were supported with results from a mesocosm experiment. Root phosphatase activity of plants increased with species richness in the mesocosms, which was associated with greater community P and N uptake. The most prominent alien grass species of the region, Melinis minutiflora, benefited most from the higher N and P availability in the species mixtures. Hence, this study provides a novel explanation of why species-richness may sometimes promote rather than impede invasion, and highlights the need to perform facilitation experiments in multi-species communities.


Sujet(s)
Espèce introduite/statistiques et données numériques , Dispersion des plantes/physiologie , Poaceae/physiologie , Brésil , Phosphoric monoester hydrolases/métabolisme , Protéines végétales/métabolisme , Racines de plante/métabolisme , Sol/composition chimique , Microbiologie du sol , Processus stochastiques
14.
Biochem Soc Trans ; 48(3): 1035-1046, 2020 06 30.
Article de Anglais | MEDLINE | ID: mdl-32490527

RÉSUMÉ

Cardiolipin (CL) and its precursor phosphatidylglycerol (PG) are important anionic phospholipids widely distributed throughout all domains of life. They have key roles in several cellular processes by shaping membranes and modulating the activity of the proteins inserted into those membranes. They are synthesized by two main pathways, the so-called eukaryotic pathway, exclusively found in mitochondria, and the prokaryotic pathway, present in most bacteria and archaea. In the prokaryotic pathway, the first and the third reactions are catalyzed by phosphatidylglycerol phosphate synthase (Pgps) belonging to the transferase family and cardiolipin synthase (Cls) belonging to the hydrolase family, while in the eukaryotic pathway, those same reactions are catalyzed by unrelated homonymous enzymes: Pgps of the hydrolase family and Cls of the transferase family. Because of the enzymatic arrangement found in both pathways, it seems that the eukaryotic pathway evolved by convergence to the prokaryotic pathway. However, since mitochondria evolved from a bacterial endosymbiont, it would suggest that the eukaryotic pathway arose from the prokaryotic pathway. In this review, it is proposed that the eukaryote pathway evolved directly from a prokaryotic pathway by the neofunctionalization of the bacterial enzymes. Moreover, after the eukaryotic radiation, this pathway was reshaped by horizontal gene transfers or subsequent endosymbiotic processes.


Sujet(s)
Archéobactéries/enzymologie , Bactéries/enzymologie , Cardiolipides/biosynthèse , Eucaryotes/enzymologie , Phosphatidylglycérol/métabolisme , Sites de fixation , Voies de biosynthèse , Catalyse , Évolution moléculaire , Transfert horizontal de gène , Hydrolases/métabolisme , Mitochondries/métabolisme , Modèles moléculaires , Phospholipides/métabolisme , Phosphoric monoester hydrolases/métabolisme , Phylogenèse
15.
J Chem Inf Model ; 60(2): 1019-1027, 2020 02 24.
Article de Anglais | MEDLINE | ID: mdl-31967472

RÉSUMÉ

Inositol polyphosphate 5-phosphatase (OCRL-1) participates in the regulation of multiple cellular processes, through the conversion of phosphatidylinositol 4,5-phosphate to phosphatidylinositol 4-phosphate. Mutations in this protein are related to Lowe syndrome (LS) and Dent-2 disease. In this study, the impact of Lowe syndrome mutations on the interactions of OCRL-1 with other proteins was evaluated through bioinformatic and computational approaches. In the functional analysis of the interaction network of the proteins, we found that the terms of gene ontology (GO) of greater significance were related to the intracellular transport of proteins, the signal transduction mediated by small G proteins and vesicles associated with the Golgi apparatus. From the proteins present in the GO terms of greater significance Rab8a was selected because its interaction facilitates the intracellular distribution of OCRL-1. The mutation p.Asn591Lys, present in the interaction domain of OCRL-1 and Rab8a, was studied using molecular dynamics. The molecular dynamics analysis showed that the presence of this mutation causes changes in the positional fluctuations of the amino acids and affects the flexibility of the protein making the interaction with Rab8a weaker. Rab proteins establish some specific interactions, which are important for the intracellular localization of OCRL-1; therefore, our findings suggest that the phenotype observed in patients with LS, in this case, is due to the destabilizing effect of p.Asn591Lys affecting the localization of OCRL-1 and indirectly its 5-phosphatase activity in the Golgi apparatus, endosomes, and cilia.


Sujet(s)
Simulation de dynamique moléculaire , Mutation , Syndrome de Lowe/génétique , Phosphoric monoester hydrolases/génétique , Phosphoric monoester hydrolases/métabolisme , Cartographie d'interactions entre protéines , Substitution d'acide aminé , Liaison hydrogène , Phosphoric monoester hydrolases/composition chimique , Conformation des protéines , Thermodynamique , Protéines G rab/métabolisme
16.
Plant Mol Biol ; 102(3): 323-337, 2020 Feb.
Article de Anglais | MEDLINE | ID: mdl-31900819

RÉSUMÉ

KEY MESSAGE: There is a link between PAP/SAL retrograde pathway, ethylene signaling and Fe metabolism in Arabidopsis. Nuclear gene expression is regulated by a diversity of retrograde signals that travel from organelles to the nucleus in a lineal or classical model. One such signal molecule is 3'-phosphoadenisine-5'-phosphate (PAP) and it's in vivo levels are regulated by SAL1/FRY1, a phosphatase enzyme located in chloroplast and mitochondria. This metabolite inhibits the action of a group of exorribonucleases which participate in post-transcriptional gene expression regulation. Transcriptome analysis of Arabidopsis thaliana mutant plants in PAP-SAL1 pathway revealed that the ferritin genes AtFER1, AtFER3, and AtFER4 are up-regulated. In this work we studied Fe metabolism in three different mutants of the PAP/SAL1 retrograde pathway. Mutant plants showed increased Fe accumulation in roots, shoots and seeds when grown in Fe-sufficient condition, and a constitutive activation of the Strategy I Fe uptake genes. As a consequence, they grew more vigorously than wild type plants in Fe-deficient medium. However, when mutant plants grown in Fe-deficient conditions were sprayed with Fe in their leaves, they were unable to deactivate root Fe uptake. Ethylene synthesis inhibition revert the constitutive Fe uptake phenotype. We propose that there is a link between PAP/SAL pathway, ethylene signaling and Fe metabolism.


Sujet(s)
ADP/métabolisme , Protéines d'Arabidopsis/métabolisme , Arabidopsis/métabolisme , Fer/métabolisme , Phosphoric monoester hydrolases/métabolisme , Transduction du signal , ADP/génétique , Arabidopsis/génétique , Arabidopsis/croissance et développement , Protéines d'Arabidopsis/génétique , Chlorophylle , Chloroplastes/métabolisme , Ferritines/génétique , Ferritines/métabolisme , Analyse de profil d'expression de gènes , Régulation de l'expression des gènes végétaux , Gènes de plante/génétique , Homéostasie , Mitochondries/métabolisme , Mutation , Phosphoric monoester hydrolases/génétique , Feuilles de plante/métabolisme , Racines de plante/croissance et développement , Racines de plante/métabolisme
17.
Infect Immun ; 88(2)2020 01 22.
Article de Anglais | MEDLINE | ID: mdl-31740531

RÉSUMÉ

l-Serine is a nonessential amino acid and a key intermediate in several relevant metabolic pathways. In bacteria, the major source of l-serine is the phosphorylated pathway, which comprises three enzymes: d-3-phosphoglycerate dehydrogenase (PGDH; SerA), phosphoserine amino transferase (PSAT; SerC), and l-phosphoserine phosphatase (PSP; SerB). The Brucella abortus genome encodes two PGDHs (SerA-1 and SerA-2), involved in the first step in l-serine biosynthesis, and one PSAT and one PSP, responsible for the second and third steps, respectively. In this study, we demonstrate that the serA1 serA2 double mutant and the serC and serB single mutants are auxotrophic for l-serine. These auxotrophic mutants can be internalized but are unable to replicate in HeLa cells and in J774A.1 macrophage-like cells. Replication defects of auxotrophic mutants can be reverted by cell medium supplementation with l-serine at early times postinfection. In addition, the serB mutant is attenuated in the murine intraperitoneal infection model and has an altered lipid composition, since the lack of l-serine abrogates phosphatidylethanolamine synthesis in this strain. Taken together, these results reveal that limited availability of l-serine within the host cell impairs proliferation of the auxotrophic strains, highlighting the relevance of this biosynthetic pathway in Brucella pathogenicity.


Sujet(s)
Brucella abortus/croissance et développement , Brucella abortus/métabolisme , Prolifération cellulaire/physiologie , Sérine/métabolisme , Animaux , Voies de biosynthèse/physiologie , Lignée cellulaire tumorale , Femelle , Cellules HeLa , Humains , Macrophages/métabolisme , Macrophages/microbiologie , Voies et réseaux métaboliques/physiologie , Souris , Souris de lignée BALB C , Phosphoric monoester hydrolases/métabolisme , Phosphorylation/physiologie
18.
Sci Rep ; 9(1): 11923, 2019 08 15.
Article de Anglais | MEDLINE | ID: mdl-31417135

RÉSUMÉ

Cryptococcosis is a fungal disease caused by C. neoformans. To adapt and survive in diverse ecological niches, including the animal host, this opportunistic pathogen relies on its ability to uptake nutrients, such as carbon, nitrogen, iron, phosphate, sulfur, and amino acids. Genetic circuits play a role in the response to environmental changes, modulating gene expression and adjusting the microbial metabolism to the nutrients available for the best energy usage and survival. We studied the sulfur amino acid biosynthesis and its implications on C. neoformans biology and virulence. CNAG_04798 encodes a BZip protein and was annotated as CYS3, which has been considered an essential gene. However, we demonstrated that CYS3 is not essential, in fact, its knockout led to sulfur amino acids auxotroph. Western blots and fluorescence microscopy indicated that GFP-Cys3, which is expressed from a constitutive promoter, localizes to the nucleus in rich medium (YEPD); the addition of methionine and cysteine as sole nitrogen source (SD-N + Met/Cys) led to reduced nuclear localization and protein degradation. By proteomics, we identified and confirmed physical interaction among Gpp2, Cna1, Cnb1 and GFP-Cys3. Deletion of the calcineurin and GPP2 genes in a GFP-Cys3 background demonstrated that calcineurin is required to maintain Cys3 high protein levels in YEPD and that deletion of GPP2 causes GFP-Cys3 to persist in the presence of sulfur amino acids. Global transcriptional profile of mutant and wild type by RNAseq revealed that Cys3 controls all branches of the sulfur amino acid biosynthesis, and sulfur starvation leads to induction of several amino acid biosynthetic routes. In addition, we found that Cys3 is required for virulence in Galleria mellonella animal model.


Sujet(s)
Acides aminés soufrés/biosynthèse , Voies de biosynthèse , Calcineurine/métabolisme , Cryptococcus neoformans/métabolisme , Protéines fongiques/métabolisme , Phosphoric monoester hydrolases/métabolisme , Voies de biosynthèse/génétique , Cryptococcus neoformans/génétique , Cryptococcus neoformans/croissance et développement , Cryptococcus neoformans/pathogénicité , Régulation de l'expression des gènes fongiques , Gene Ontology , Protéines à fluorescence verte/métabolisme , Modèles biologiques , État nutritionnel , Transport des protéines , Protéomique , Soufre/métabolisme , Facteurs de transcription/métabolisme , Transcription génétique , Virulence/génétique
19.
Molecules ; 24(13)2019 Jul 07.
Article de Anglais | MEDLINE | ID: mdl-31284667

RÉSUMÉ

Salinity stress limited the production in over 30% of irrigated crops and 7% of dryland agriculture worldwide. The objective was to evaluate the effects of NaCl-stress on the enzymatic activity in tomato. Two experiments were carried out in germination and early vegetative growth stages. The activity of proline and peroxidase of eight varieties (Missouri, Yaqui, Vita, Feroz, Rio Grande, Tropic, Ace, and Floradade) submitted to NaCl concentrations (0, 50, 100, 150 and 200 mM de NaCl) and the semi-quantitative activity of 19 enzymes APY ZYM® were measured under a completely randomized design with four replications. Data were analyzed using univariate-multivariate analysis of variance, Tukey's HSD (p = 0.05), canonical discriminant and cluster analysis. The results showed significant differences between varieties and NaCl in proline content. Proline increased as the NaCl concentration increased. Peroxidase did no show significant differences. Eight enzymes were included within the model to properly classify the varieties and NaCl. In shoots, varieties and NaCl showed that enzymatic activity was higher in the order of alkaline-phosphatase > leucine arylamidase > acid phosphatase > naphthol-AS-BI-phosphohydrolase > n-acetyl-ß-glucosaminidase > ß-galactosidase, while in roots was higher in the order of alkaline-phosphatase > naphthol-AS-BI-phosphohydrolase > acid phosphatase > n-acetyl-ß-glucosaminidase. Acid and alkali phosphatase, lipase, esterase, ß-galactosidase, and trypsin can be a potential biomarker for NaCl-stress tolerance in tomato.


Sujet(s)
Esterases/métabolisme , N-Glycosyl hydrolases/métabolisme , Peptide hydrolases/métabolisme , Phosphoric monoester hydrolases/métabolisme , Tolérance au sel , Chlorure de sodium/pharmacologie , Solanum lycopersicum/effets des médicaments et des substances chimiques , Solanum lycopersicum/physiologie , Marqueurs biologiques , Analyse de regroupements , Activation enzymatique , Pousses de plante/physiologie , Proline/métabolisme , Protéome , Protéomique , Plant/physiologie
20.
Chem Biol Interact ; 308: 323-331, 2019 Aug 01.
Article de Anglais | MEDLINE | ID: mdl-31173750

RÉSUMÉ

Organophosphorus compounds have been widely employed to the development of warfare nerve agents and pesticides, resulting in a huge number of people intoxicated annually, being a serious problem of public health. Efforts worldwide have been done in order to design new technologies that are capable of combating or even reversing the poisoning caused by these OP nerve agents. In this line, the bioremediation arises as a promising and efficient alternative for this purpose. As an example of degrading enzymes, there is the organophosphate-degrading (OpdA) enzyme from Agrobacterium radiobacter, which has been quite investigated experimentally due to its high performance in the degradation of neurotoxic nerve agents. This work aims to look into the structural and electronic details that govern the interaction modes of these compounds in the OpdA active site, with the posterior hydrolysis reaction prediction. Our findings have brought about data about the OpdA performance towards different nerve agents, and among them, we may realize that the degradation efficiency strongly depends on the nerve agent structure and its stereochemistry, being in this case the compound Tabun the one more effectively hydrolyzed. By means of the chemical bonds (AIM) and orbitals (FERMO) analysis, it is suggested that the initial reactivity of the OP nerve agents in the OpdA active site does not necessarily dictate the reactivity and interaction modes over the reaction coordinate.


Sujet(s)
Dépollution biologique de l'environnement , Agents neurotoxiques/métabolisme , Agrobacterium tumefaciens/enzymologie , Protéines bactériennes/composition chimique , Protéines bactériennes/métabolisme , Sites de fixation , Biocatalyse , Domaine catalytique , Humains , Simulation de docking moléculaire , Agents neurotoxiques/composition chimique , Phosphoric monoester hydrolases/composition chimique , Phosphoric monoester hydrolases/métabolisme , Théorie quantique , Sarin/composition chimique , Sarin/métabolisme
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