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1.
Phytopathology ; 113(9): 1745-1760, 2023 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-37885045

RESUMEN

The success of virus transmission by vectors relies on intricate trophic interactions between three partners, the host plant, the virus, and the vector. Despite numerous studies that showed the capacity of plant viruses to manipulate their host plant to their benefit, and potentially of their transmission, the molecular mechanisms sustaining this phenomenon has not yet been extensively analyzed at the molecular level. In this study, we focused on the deregulations induced in Arabidopsis thaliana by an aphid vector that were alleviated when the plants were infected with turnip yellows virus (TuYV), a polerovirus strictly transmitted by aphids in a circulative and nonpropagative mode. By setting up an experimental design mimicking the natural conditions of virus transmission, we analyzed the deregulations in plants infected with TuYV and infested with aphids by a dual transcriptomic and metabolomic approach. We observed that the virus infection alleviated most of the gene deregulations induced by the aphids in a noninfected plant at both time points analyzed (6 and 72 h) with a more pronounced effect at the later time point of infestation. The metabolic composition of the infected and infested plants was altered in a way that could be beneficial for the vector and the virus transmission. Importantly, these substantial modifications observed in infected and infested plants correlated with a higher TuYV transmission efficiency. This study revealed the capacity of TuYV to alter the plant nutritive content and the defense reaction against the aphid vector to promote the viral transmission.


Asunto(s)
Áfidos , Arabidopsis , Luteoviridae , Virus de Plantas , Animales , Enfermedades de las Plantas , Insectos Vectores , Arabidopsis/genética , Luteoviridae/fisiología
2.
Neurobiol Dis ; 127: 242-252, 2019 07.
Artículo en Inglés | MEDLINE | ID: mdl-30872158

RESUMEN

Niemann-Pick type C disease (NPCD) is a neurodegenerative disease associated with increases in cellular cholesterol and glycolipids and most commonly caused by defective NPC1, a late endosomal protein. Using ratiometric probes we find that NPCD cells show increased endolysosomal pH. In addition U18666A, an inhibitor of NPC1, was found to increase endolysosomal pH, and the number, size and heterogeneity of endolysosomal vesicles. NPCD fibroblasts and cells treated with U18666A also show disrupted targeting of fluorescent lipid BODIPY-LacCer to high pH vesicles. Inhibiting non-lysosomal glucocerebrosidase (GBA2) reversed increases in endolysosomal pH and restored disrupted BODIPY-LacCer trafficking in NPCD fibroblasts. GBA2 KO cells also show decreased endolysosomal pH. NPCD fibroblasts also show increased expression of a key subunit of the lysosomal proton pump vATPase on GBA2 inhibition. The results are consistent with a model where both endolysosomal pH and Golgi targeting of BODIPY-LacCer are dependent on adequate levels of cytosolic-facing GlcCer, which are reduced in NPC disease.


Asunto(s)
Citosol/metabolismo , Endosomas/metabolismo , Glucosilceramidas/metabolismo , Lisosomas/metabolismo , Enfermedad de Niemann-Pick Tipo C/metabolismo , Androstenos/farmacología , Animales , Citosol/efectos de los fármacos , Endosomas/efectos de los fármacos , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Humanos , Lisosomas/efectos de los fármacos , Ratones , Proteína Niemann-Pick C1/antagonistas & inhibidores
3.
Int J Mol Sci ; 19(8)2018 Aug 07.
Artículo en Inglés | MEDLINE | ID: mdl-30087282

RESUMEN

Aphids are important pests which cause direct damage by feeding or indirect prejudice by transmitting plant viruses. Viruses are known to induce modifications of plant cues in ways that can alter vector behavior and virus transmission. In this work, we addressed whether the modifications induced by the aphid-transmitted Turnip yellows virus (TuYV) in the model plant Arabidopsis thaliana also apply to the cultivated plant Camelina sativa, both belonging to the Brassicaceae family. In most experiments, we observed a significant increase in the relative emission of volatiles from TuYV-infected plants. Moreover, due to plant size, the global amounts of volatiles emitted by C. sativa were higher than those released by A. thaliana. In addition, the volatiles released by TuYV-infected C. sativa attracted the TuYV vector Myzus persicae more efficiently than those emitted by non-infected plants. In contrast, no such preference was observed for A. thaliana. We propose that high amounts of volatiles rather than specific metabolites are responsible for aphid attraction to infected C. sativa. This study points out that the data obtained from the model pathosystem A. thaliana/TuYV cannot be straightforwardly extrapolated to a related plant species infected with the same virus.


Asunto(s)
Áfidos/virología , Brassica/virología , Herbivoria , Insectos Vectores/virología , Enfermedades de las Plantas/virología , Virus de Plantas/aislamiento & purificación , Animales , Áfidos/fisiología , Arabidopsis/fisiología , Arabidopsis/virología , Brassica/fisiología , Insectos Vectores/fisiología , Compuestos Orgánicos Volátiles/análisis , Compuestos Orgánicos Volátiles/metabolismo
4.
New Phytol ; 213(1): 264-274, 2017 01.
Artículo en Inglés | MEDLINE | ID: mdl-27560385

RESUMEN

Monoterpenes are important constituents of the aromas of food and beverages, including wine. Among monoterpenes in wines, wine lactone has the most potent odor. It was proposed to form via acid-catalyzed cyclization of (E)-8-carboxylinalool during wine maturation. It only reaches very low concentrations in wine but its extremely low odor detection threshold makes it an important aroma compound. Using LC-MS/MS, we show here that the (E)-8-carboxylinalool content in wines correlates with their wine lactone content and estimate the kinetic constant for the very slow formation of wine lactone from (E)-8-carboxylinalool. We show that (E)-8-carboxylinalool is accumulated as a glycoside in grape (Vitis vinifera) berries and that one of the cytochrome P450 enzymes most highly expressed in maturing berries, CYP76F14, efficiently oxidizes linalool to (E)-8-carboxylinalool. Our analysis of (E)-8-carboxylinalool in Riesling × Gewurztraminer grapevine progeny established that the CYP76F14 gene co-locates with a quantitative trait locus for (E)-8-carboxylinalool content in grape berries. Our data support the role of CYP76F14 as the major (E)-8-carboxylinalool synthase in grape berries and the role of (E)-8-carboxylinalool as a precursor to wine lactone in wine, providing new insights into wine and grape aroma metabolism, and new methods for food and aroma research and production.


Asunto(s)
Sistema Enzimático del Citocromo P-450/metabolismo , Lactonas/metabolismo , Odorantes/análisis , Vitis/enzimología , Vino/análisis , Monoterpenos Acíclicos , Frutas/enzimología , Frutas/genética , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Lactonas/química , Monoterpenos/química , Monoterpenos/metabolismo , Hojas de la Planta/metabolismo , Proteínas de Plantas/metabolismo , Sitios de Carácter Cuantitativo/genética , Nicotiana/metabolismo , Vitis/genética
5.
Environ Microbiol ; 18(4): 1289-300, 2016 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-26769162

RESUMEN

Arsenic is a toxic metalloid known to generate an important oxidative stress in cells. In the present study, we focused our attention on an alga related to the genus Coccomyxa, exhibiting an extraordinary capacity to resist high concentrations of arsenite and arsenate. The integrated analysis of high-throughput transcriptomic data and non-targeted metabolomic approaches highlighted multiple levels of protection against arsenite. Indeed, Coccomyxa sp. Carn induced a set of transporters potentially preventing the accumulation of this metalloid in the cells and presented a distinct arsenic metabolism in comparison to another species more sensitive to that compound, i.e. Euglena gracilis, especially in regard to arsenic methylation. Interestingly, Coccomyxa sp. Carn was characterized by a remarkable accumulation of the strong antioxidant glutathione (GSH). Such observation could explain the apparent low oxidative stress in the intracellular compartment, as suggested by the transcriptomic analysis. In particular, the high amount of GSH in the cell could play an important role for the tolerance to arsenate, as suggested by its partial oxidation into oxidized glutathione in presence of this metalloid. Our results therefore reveal that this alga has acquired multiple and original defence mechanisms allowing the colonization of extreme ecosystems such as acid mine drainages.


Asunto(s)
Arseniatos/metabolismo , Arsenitos/metabolismo , Chlorophyta/metabolismo , Glutatión/metabolismo , Metabolómica , Proteínas de Transporte de Membrana/metabolismo , Metilación , Oxidación-Reducción
6.
Environ Microbiol ; 17(6): 1941-9, 2015 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-24698441

RESUMEN

Arsenic is a toxic metalloid known to cause multiple and severe cellular damages, including lipid peroxidation, protein misfolding, mutagenesis and double and single-stranded DNA breaks. Thus, exposure to this compound is lethal for most organisms but some species such as the photosynthetic protist Euglena mutabilis are able to cope with very high concentrations of this metalloid. Our comparative transcriptomic approaches performed on both an arsenic hypertolerant protist, i.e. E. mutabilis, and a more sensitive one, i.e. E. gracilis, revealed multiple mechanisms involved in arsenic tolerance. Indeed, E. mutabilis prevents efficiently the accumulation of arsenic in the cell through the expression of several transporters. More surprisingly, this protist induced the expression of active DNA reparation and protein turnover mechanisms, which allow E. mutabilis to maintain functional integrity of the cell under challenging conditions. Our observations suggest that this protist has acquired specific functions regarding arsenic and has developed an original metabolism to cope with acid mine drainages-related stresses.


Asunto(s)
Arsénico/metabolismo , Transporte Biológico/genética , Euglena/metabolismo , Proteínas de Transporte de Membrana/genética , Transporte Biológico/fisiología , Resistencia a Medicamentos/genética , Resistencia a Medicamentos/fisiología , Euglena/efectos de los fármacos , Euglena/genética , Proteínas de Transporte de Membrana/metabolismo , Fotosíntesis
7.
J Exp Bot ; 66(11): 3243-57, 2015 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-25873669

RESUMEN

Stilbenes, as important secondary metabolites of grapevine, represent central phytoalexins and therefore constitute an important element of basal immunity. In this study, potential genetic variation in Vitis vinifera ssp. sylvestris, the ancestor of cultivated grapevine, was sought with respect to their output of stilbenes and potential use for resistance breeding. Considerable variation in stilbene inducibility was identified in V. vinifera ssp. sylvestris. Genotypic differences in abundance and profiles of stilbenes that are induced in response to a UV-C pulse are shown. Two clusters of stilbene 'chemovars' emerged: one cluster showed quick and strong accumulation of stilbenes, almost exclusively in the form of non-glycosylated resveratrol and viniferin, while the second cluster accumulated fewer stilbenes and relatively high proportions of piceatannol and the glycosylated piceid. For all 86 genotypes, a time dependence of the stilbene pattern was observed: piceid, resveratrol, and piceatannol accumulated earlier, whereas the viniferins were found later. It was further observed that the genotypic differences in stilbene accumulation were preceded by differential accumulation of the transcripts for chalcone synthase (CHS) and stilbene-related genes: phenylalanine ammonium lyase (PAL), stilbene synthase (StSy), and resveratrol synthase (RS). A screen of the population with respect to susceptibility to downy mildew of grapevine (Plasmopara viticola) revealed considerable variability. The subpopulation of genotypes with high stilbene inducibility was significantly less susceptible as compared with low-stilbene genotypes, and for representative genotypes it could be shown that the inducibility of stilbene synthase by UV correlated with the inducibility by the pathogen.


Asunto(s)
Variación Genética , Enfermedades de las Plantas/inmunología , Estilbenos/metabolismo , Vitis/genética , Aciltransferasas/genética , Aciltransferasas/metabolismo , Regulación de la Expresión Génica de las Plantas , Genotipo , Glucósidos/metabolismo , Oomicetos/fisiología , Inmunidad de la Planta , Hojas de la Planta/genética , Hojas de la Planta/inmunología , Hojas de la Planta/metabolismo , Hojas de la Planta/efectos de la radiación , Estomas de Plantas/genética , Estomas de Plantas/inmunología , Estomas de Plantas/metabolismo , Estomas de Plantas/efectos de la radiación , Resveratrol , Sesquiterpenos/metabolismo , Rayos Ultravioleta , Vitis/inmunología , Vitis/metabolismo , Vitis/efectos de la radiación , Fitoalexinas
8.
Metabolites ; 13(1)2023 Jan 02.
Artículo en Inglés | MEDLINE | ID: mdl-36676996

RESUMEN

Alternaria leaf blight, caused by the fungus Alternaria dauci, is the most damaging foliar disease of carrot. Some carrot genotypes exhibit partial resistance to this pathogen and resistance Quantitative Trait Loci (rQTL) have been identified. Co-localization of metabolic QTL and rQTL identified camphene, α-pinene, α-bisabolene, ß-cubebene, caryophyllene, germacrene D and α-humulene as terpenes potentially involved in carrot resistance against ALB. By combining genomic and transcriptomic analyses, we identified, under the co-localization regions, terpene-related genes which are differentially expressed between a resistant and a susceptible carrot genotype. These genes include five terpene synthases and twenty transcription factors. In addition, significant mycelial growth inhibition was observed in the presence of α-humulene and caryophyllene.

9.
J Cell Biol ; 179(1): 101-15, 2007 Oct 08.
Artículo en Inglés | MEDLINE | ID: mdl-17923531

RESUMEN

Glycosphingolipids are controlled by the spatial organization of their metabolism and by transport specificity. Using immunoelectron microscopy, we localize to the Golgi stack the glycosyltransferases that produce glucosylceramide (GlcCer), lactosylceramide (LacCer), and GM3. GlcCer is synthesized on the cytosolic side and must translocate across to the Golgi lumen for LacCer synthesis. However, only very little natural GlcCer translocates across the Golgi in vitro. As GlcCer reaches the cell surface when Golgi vesicular trafficking is inhibited, it must translocate across a post-Golgi membrane. Concanamycin, a vacuolar proton pump inhibitor, blocks translocation independently of multidrug transporters that are known to translocate short-chain GlcCer. Concanamycin did not reduce LacCer and GM3 synthesis. Thus, GlcCer destined for glycolipid synthesis follows a different pathway and transports back into the endoplasmic reticulum (ER) via the late Golgi protein FAPP2. FAPP2 knockdown strongly reduces GM3 synthesis. Overall, we show that newly synthesized GlcCer enters two pathways: one toward the noncytosolic surface of a post-Golgi membrane and one via the ER toward the Golgi lumen LacCer synthase.


Asunto(s)
Glucosilceramidas/metabolismo , Glicoesfingolípidos/biosíntesis , Aparato de Golgi/metabolismo , Animales , Antígenos CD/metabolismo , Transporte Biológico/efectos de los fármacos , Transporte Biológico/fisiología , Brefeldino A/farmacología , Proteínas Portadoras/metabolismo , Proteínas Portadoras/fisiología , Bovinos , Línea Celular , Cricetinae , Cricetulus , Retículo Endoplásmico/metabolismo , Glicosiltransferasas/metabolismo , Aparato de Golgi/enzimología , Humanos , Membranas Intracelulares/metabolismo , Lactosilceramidos/metabolismo , Macrólidos/farmacología , Ratones , Microscopía Fluorescente , Modelos Biológicos , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas Serina-Treonina Quinasas/fisiología , Inhibidores de la Bomba de Protones/farmacología , Ratas
10.
Appl Microbiol Biotechnol ; 93(4): 1735-44, 2012 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-21792588

RESUMEN

Euglena mutabilis is a protist ubiquitously found in extreme environments such as acid mine drainages which are often rich in arsenic. The response of E. mutabilis to this metalloid was compared to that of Euglena gracilis, a protist not found in such environments. Membrane fatty acid composition, cell surface properties, arsenic accumulation kinetics, and intracellular arsenic speciation were determined. The results revealed a modification in fatty acid composition leading to an increased membrane fluidity in both Euglena species under sublethal arsenic concentrations exposure. This increased membrane fluidity correlated to an induced gliding motility observed in E. mutabilis in the presence of this metalloid but did not affect the flagellar dependent motility of E. gracilis. Moreover, when compared to E. gracilis, E. mutabilis showed highly hydrophobic cell surface properties and a higher tolerance to water-soluble arsenical compounds but not to hydrophobic ones. Finally, E. mutabilis showed a lower accumulation of total arsenic in the intracellular compartment and an absence of arsenic methylated species in contrast to E. gracilis. Taken together, our results revealed the existence of a specific arsenical response of E. mutabilis that may play a role in its hypertolerance to this toxic metalloid.


Asunto(s)
Adaptación Fisiológica , Arsénico/toxicidad , Euglena/efectos de los fármacos , Contaminantes del Suelo/toxicidad , Membrana Celular/química , Membrana Celular/efectos de los fármacos , Tolerancia a Medicamentos , Euglena/química , Euglena/fisiología , Ácidos Grasos/análisis , Interacciones Hidrofóbicas e Hidrofílicas , Locomoción , Fluidez de la Membrana/efectos de los fármacos , Propiedades de Superficie
11.
Appl Environ Microbiol ; 77(13): 4685-92, 2011 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-21571879

RESUMEN

In this study, new strains were isolated from an environment with elevated arsenic levels, Sainte-Marie-aux-Mines (France), and the diversity of aoxB genes encoding the arsenite oxidase large subunit was investigated. The distribution of bacterial aoxB genes is wider than what was previously thought. AoxB subfamilies characterized by specific signatures were identified. An exhaustive analysis of AoxB sequences from this study and from public databases shows that horizontal gene transfer has likely played a role in the spreading of aoxB in prokaryotic communities.


Asunto(s)
Archaea/enzimología , Archaea/metabolismo , Arsenitos/metabolismo , Bacterias/enzimología , Bacterias/metabolismo , Microbiología Ambiental , Oxidorreductasas/genética , Archaea/genética , Archaea/aislamiento & purificación , Bacterias/genética , Bacterias/aislamiento & purificación , Análisis por Conglomerados , ADN de Archaea/química , ADN de Archaea/genética , ADN Bacteriano/química , ADN Bacteriano/genética , Francia , Datos de Secuencia Molecular , Oxidación-Reducción , Filogenia , Análisis de Secuencia de ADN , Homología de Secuencia
12.
IEEE J Biomed Health Inform ; 25(2): 381-392, 2021 02.
Artículo en Inglés | MEDLINE | ID: mdl-32750943

RESUMEN

There is a need for a reliable and reproducible quantification of the immune infiltrate within the heterogeneous microenvironment of tumors in order to support therapy selection in oncology. Here we present an automated, modular method for whole-slide image analysis of the spatial distribution of tumor-infiltrating CD8-positive lymphocytes. The method uses a deep learning tissue-type classification algorithm on the hematoxylin eosin (HE) stained tissue section to identify the central tumor (CT) and invasive margin (IM) of the tumor. A CD8-positive cell detection algorithm using a deep learning-based nucleus detection is applied to a sequential immunohistochemistry (IHC)-stained tissue section. Image registration then allows obtaining IHC-derived CD8 scores for the HE-derived CT and the IM, respectively. Both, the mean and the standard deviation of the spatial CD8-positive density distributions were determined for the CT and IM in a cohort of post-menopausal, estrogen receptor-positive invasive breast cancer patients who received adjuvant tamoxifen therapy. Spatial density distributions were found to be highly heterogeneous. In contrast to previous studies, CD8 density in the IM and CT correlated positively with clinical outcome. However, statistical significance was only achieved for the standard deviation of the CD8 density distribution. We hypothesize that this is due to the positive contribution of local high-density areas. The IM/CT density ratio did not correlate with outcome. In view of the clinical relevance of our finding, we would like to encourage a study with a larger cohort. Our modular pipeline approach allows a robust and objective scoring of CD8 infiltrate based on routine pathology staining and should contribute to clinical adoption of computational pathology.


Asunto(s)
Neoplasias de la Mama , Neoplasias de la Mama/diagnóstico por imagen , Neoplasias de la Mama/tratamiento farmacológico , Linfocitos T CD8-positivos , Femenino , Humanos , Procesamiento de Imagen Asistido por Computador , Inmunohistoquímica , Linfocitos Infiltrantes de Tumor , Microambiente Tumoral
13.
Front Plant Sci ; 9: 360, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-29619037

RESUMEN

The Oomycete Plasmopara viticola is responsible for downy mildew, which is one of the most damaging grapevine diseases. Due to the strictly biotrophic way of life of P. viticola, its metabolome is relatively poorly characterized. In this work, we have used a mass spectrometry-based non-targeted metabolomic approach to identify potential Plasmopara-specific metabolites. This has led to the characterization and structural elucidation of compounds belonging to three families of atypical lipids, which are not detected in healthy grapevine tissues. These lipids include ceramides and derivatives of arachidonic and eicosapentaenoic acid, most of which had not been previously described in Oomycetes. Furthermore, we show that these lipids can be detected in Plasmopara-infected tissues at very early stages of the infection process, long before the appearance the first visible symptoms of the disease. Therefore, the potential use of these specific lipids as markers to monitor the development of P. viticola is discussed.

14.
Sci Rep ; 8(1): 13746, 2018 09 13.
Artículo en Inglés | MEDLINE | ID: mdl-30213972

RESUMEN

Alternaria Leaf Blight (ALB), caused by the fungus Alternaria dauci, is the most damaging foliar disease affecting carrots (Daucus carota). In order to identify compounds potentially linked to the resistance to A. dauci, we have used a combination of targeted and non-targeted metabolomics to compare the leaf metabolome of four carrot genotypes with different resistance levels. Targeted analyses were focused on terpene volatiles, while total leaf methanolic extracts were subjected to non-targeted analyses using liquid chromatography couple to high-resolution mass spectrometry. Differences in the accumulation of major metabolites were highlighted among genotypes and some of these metabolites were identified as potentially involved in resistance or susceptibility. A bulk segregant analysis on F3 progenies obtained from a cross between one of the resistant genotypes and a susceptible one, confirmed or refuted the hypothesis that the metabolites differentially accumulated by these two parents could be linked to resistance.


Asunto(s)
Alternaria/metabolismo , Daucus carota/metabolismo , Hojas de la Planta/metabolismo , Alternaria/química , Daucus carota/genética , Genotipo , Metaboloma/genética , Enfermedades de las Plantas/genética , Hojas de la Planta/química , Metabolismo Secundario/genética
15.
FEBS Lett ; 580(4): 1171-7, 2006 Feb 13.
Artículo en Inglés | MEDLINE | ID: mdl-16376334

RESUMEN

Many mammalian ABC transporters move membrane lipids to acceptor lipid assemblies in the extracellular aqueous milieu. Because the desorption from the membrane costs more energy than provided by two ATPs, the transporter probably only translocates the lipid to a partially hydrophilic site on its extracellular face. From this high-energy site, the lipid may efficiently move to the acceptor, which ideally is bound to the transporter, or, in the absence of an acceptor, fall back into the membrane. If the lipid originated from the cytosolic membrane surface, this represents lipid flop and is probably a side activity of the transporters.


Asunto(s)
Transportadoras de Casetes de Unión a ATP/metabolismo , Metabolismo de los Lípidos , Humanos , Recién Nacido , Errores Innatos del Metabolismo Lipídico/metabolismo
16.
Viruses ; 8(11)2016 11 19.
Artículo en Inglés | MEDLINE | ID: mdl-27869783

RESUMEN

With the increasing availability of aphid genomic data, it is necessary to develop robust functional validation methods to evaluate the role of specific aphid genes. This work represents the first study in which five different techniques, all based on RNA interference and on oral acquisition of double-stranded RNA (dsRNA), were developed to silence two genes, ALY and Eph, potentially involved in polerovirus transmission by aphids. Efficient silencing of only Eph transcripts, which are less abundant than those of ALY, could be achieved by feeding aphids on transgenic Arabidopsis thaliana expressing an RNA hairpin targeting Eph, on Nicotiana benthamiana infected with a Tobacco rattle virus (TRV)-Eph recombinant virus, or on in vitro-synthesized Eph-targeting dsRNA. These experiments showed that the silencing efficiency may differ greatly between genes and that aphid gut cells seem to be preferentially affected by the silencing mechanism after oral acquisition of dsRNA. In addition, the use of plants infected with recombinant TRV proved to be a promising technique to silence aphid genes as it does not require plant transformation. This work highlights the need to pursue development of innovative strategies to reproducibly achieve reduction of expression of aphid genes.


Asunto(s)
Áfidos/genética , Entomología/métodos , Técnicas de Silenciamiento del Gen/métodos , Genes de Insecto , Interferencia de ARN , Animales , Áfidos/crecimiento & desarrollo , Arabidopsis/parasitología , Nicotiana/parasitología
17.
Front Microbiol ; 6: 993, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26441922

RESUMEN

The acid mine drainage (AMD) in Carnoulès (France) is characterized by the presence of toxic metals such as arsenic. Several bacterial strains belonging to the Thiomonas genus, which were isolated from this AMD, are able to withstand these conditions. Their genomes carry several genomic islands (GEIs), which are known to be potentially advantageous in some particular ecological niches. This study focused on the role of the "urea island" present in the Thiomonas CB2 strain, which carry the genes involved in urea degradation processes. First, genomic comparisons showed that the genome of Thiomonas sp. CB2, which is able to degrade urea, contains a urea genomic island which is incomplete in the genome of other strains showing no urease activity. The urease activity of Thiomonas sp. CB2 enabled this bacterium to maintain a neutral pH in cell cultures in vitro and prevented the occurrence of cell death during the growth of the bacterium in a chemically defined medium. In AMD water supplemented with urea, the degradation of urea promotes iron, aluminum and arsenic precipitation. Our data show that ureC was expressed in situ, which suggests that the ability to degrade urea may be expressed in some Thiomonas strains in AMD, and that this urease activity may contribute to their survival in contaminated environments.

18.
PLoS One ; 9(1): e86218, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24465968

RESUMEN

Eukaryotic cells control their proteome by regulating protein production and protein clearance. Protein production is determined to a large extent by mRNA levels, whereas protein degradation depends mostly upon the proteasome. Dysfunction of the proteasome leads to the accumulation of non-functional proteins that can aggregate, be toxic for the cell, and, in extreme cases, lead to cell death. mRNA levels are controlled by their rates of synthesis and degradation. Recent evidence indicates that these rates have oppositely co-evolved to ensure appropriate mRNA levels. This opposite co-evolution has been correlated with the mutations in the Ccr4-Not complex. Consistently, the deadenylation enzymes responsible for the rate-limiting step in eukaryotic mRNA degradation, Caf1 and Ccr4, are subunits of the Ccr4-Not complex. Another subunit of this complex is a RING E3 ligase, Not4. It is essential for cellular protein solubility and has been proposed to be involved in co-translational quality control. An open question has been whether this role of Not4 resides strictly in the regulation of the deadenylation module of the Ccr4-Not complex. However, Not4 is important for proper assembly of the proteasome, and the Ccr4-Not complex may have multiple functional modules that participate in protein quality control in different ways. In this work we studied how the functions of the Caf1/Ccr4 and Not4 modules are connected. We concluded that Not4 plays a role in protein quality control independently of the Ccr4 deadenylase, and that it is involved in clearance of aberrant proteins at least in part via the proteasome.


Asunto(s)
Ribonucleasas/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/enzimología , Ubiquitina-Proteína Ligasas/metabolismo , Ácido Azetidinocarboxílico/farmacología , Catepsina A/metabolismo , Estabilidad de Enzimas/efectos de los fármacos , Eliminación de Gen , Proteínas Mutantes/metabolismo , Fenotipo , Polirribosomas/efectos de los fármacos , Polirribosomas/metabolismo , Poliubiquitina/metabolismo , Complejo de la Endopetidasa Proteasomal/metabolismo , Biosíntesis de Proteínas/efectos de los fármacos , Estructura Cuaternaria de Proteína , Proteolisis/efectos de los fármacos , Proteínas Represoras , Saccharomyces cerevisiae/efectos de los fármacos , Especificidad por Sustrato/efectos de los fármacos , Temperatura , Ubiquitinación/efectos de los fármacos
19.
ISME J ; 6(7): 1391-402, 2012 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-22237547

RESUMEN

Euglena mutabilis is a photosynthetic protist found in acidic aquatic environments such as peat bogs, volcanic lakes and acid mine drainages (AMDs). Through its photosynthetic metabolism, this protist is supposed to have an important role in primary production in such oligotrophic ecosystems. Nevertheless, the exact contribution of E. mutabilis in organic matter synthesis remains unclear and no evidence of metabolite secretion by this protist has been established so far. Here we combined in situ proteo-metabolomic approaches to determine the nature of the metabolites accumulated by this protist or potentially secreted into an AMD. Our results revealed that the secreted metabolites are represented by a large number of amino acids, polyamine compounds, urea and some sugars but no fatty acids, suggesting a selective organic matter contribution in this ecosystem. Such a production may have a crucial impact on the bacterial community present on the study site, as it has been suggested previously that prokaryotes transport and recycle in situ most of the metabolites secreted by E. mutabilis. Consequently, this protist may have an indirect but important role in AMD ecosystems but also in other ecological niches often described as nitrogen-limited.


Asunto(s)
Ecosistema , Euglena/aislamiento & purificación , Euglena/metabolismo , Minería , Ácidos/metabolismo , Francia , Metabolómica , Fotosíntesis
20.
PLoS One ; 6(8): e23181, 2011.
Artículo en Inglés | MEDLINE | ID: mdl-21876737

RESUMEN

Biofilms represent the most common microbial lifestyle, allowing the survival of microbial populations exposed to harsh environmental conditions. Here, we show that the biofilm development of a bacterial species belonging to the Thiomonas genus, frequently found in arsenic polluted sites and playing a key role in arsenic natural remediation, is markedly modified when exposed to subinhibitory doses of this toxic element. Indeed, arsenite [As(III)] exposure led to a considerable impact on biofilm maturation by strongly increasing the extracellular matrix synthesis and by promoting significant cell death and lysis within microcolonies. These events were followed by the development of complex 3D-biofilm structures and subsequently by the dispersal of remobilized cells observed inside the previously formed hollow voids. Our results demonstrate that this biofilm community responds to arsenite stress in a multimodal way, enhancing both survival and dispersal. Addressing this complex bacterial response to As(III) stress, which might be used by other microorganisms under various adverse conditions, may be essential to understand how Thiomonas strains persist in extreme environments.


Asunto(s)
Arsenitos/toxicidad , Betaproteobacteria/efectos de los fármacos , Betaproteobacteria/crecimiento & desarrollo , Betaproteobacteria/citología , Betaproteobacteria/fisiología , Biopelículas/efectos de los fármacos , Recuento de Colonia Microbiana , Matriz Extracelular/efectos de los fármacos , Matriz Extracelular/metabolismo , Movimiento/efectos de los fármacos , Ácidos Nucleicos/metabolismo , Polisacáridos Bacterianos/efectos de los fármacos , Polisacáridos Bacterianos/metabolismo
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