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91.
Miguel ángel Rubio Mauro Napolitano Jesús?A.?G. Ochoa?de?Alda Javier Santamaría-Gómez Carl J. Patterson Andrew W. Foster Roque Bru-Martínez Nigel J. Robinson Ignacio Luque 《Nucleic acids research》2015,43(20):9905-9917
Aminoacyl-tRNA synthetases (aaRSs) play a key role in deciphering the genetic message by producing charged tRNAs and are equipped with proofreading mechanisms to ensure correct pairing of tRNAs with their cognate amino acid. Duplicated aaRSs are very frequent in Nature, with 25,913 cases observed in 26,837 genomes. The oligomeric nature of many aaRSs raises the question of how the functioning and oligomerization of duplicated enzymes is organized. We characterized this issue in a model prokaryotic organism that expresses two different threonyl-tRNA synthetases, responsible for Thr-tRNAThr synthesis: one accurate and constitutively expressed (T1) and another (T2) with impaired proofreading activity that also generates mischarged Ser-tRNAThr. Low zinc promotes dissociation of dimeric T1 into monomers deprived of aminoacylation activity and simultaneous induction of T2, which is active for aminoacylation under low zinc. T2 either forms homodimers or heterodimerizes with T1 subunits that provide essential proofreading activity in trans. These findings evidence that in organisms with duplicated genes, cells can orchestrate the assemblage of aaRSs oligomers that meet the necessities of the cell in each situation. We propose that controlled oligomerization of duplicated aaRSs is an adaptive mechanism that can potentially be expanded to the plethora of organisms with duplicated oligomeric aaRSs. 相似文献
92.
Rapid detection and discrimination of fabaviruses by flow‐through hybridisation with genus‐ and species‐specific riboprobes
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I. Ferriol E. A. Rangel S. Panno S. Davino C.G. Han A. Olmos L. Rubio 《The Annals of applied biology》2015,167(1):26-35
Viruses cause significant damage in agricultural crops worldwide. Disease management requires sensitive and specific tools for virus detection and identification. Also, detection techniques need to be rapid to keep pace with the continuous emergence of new viral diseases. The genus Fabavirus is composed of five viruses infecting many economically important crops worldwide. This research describes the development of a procedure based on flow‐through hybridisation (FTH), which is faster than and as sensitive as conventional hybridisation for virus detection in tissue‐prints from infected plants. Six digoxigenin‐labelled RNA probes were synthesised with two levels of specificity: (a) five specific for each viral species within this genus, and (b) a genus‐specific probe that hybridises with a nucleotide sequence signature only found in the 5′‐untranslated region of the genus Fabavirus, which is the first of this type reported for plant viruses. The new procedure developed is useful for rapid detection and discrimination of the five fabaviruses identified so far and opens the possibility of discovering new species of this genus. 相似文献
93.
Teresa de Diego Puente Julia Gallego-Jara Sara Casta?o-Cerezo Vicente Bernal Sánchez Vanesa Fernández Espín José García de la Torre Arturo Manjón Rubio Manuel Cánovas Díaz 《The Journal of biological chemistry》2015,290(38):23077-23093
Lysine acetylation is an important post-translational modification in the metabolic regulation of both prokaryotes and eukaryotes. In Escherichia coli, PatZ (formerly YfiQ) is the only known acetyltransferase protein and is responsible for acetyl-CoA synthetase acetylation. In this study, we demonstrated PatZ-positive cooperativity in response to acetyl-CoA and the regulation of acetyl-CoA synthetase activity by the acetylation level. Furthermore, functional analysis of an E809A mutant showed that the conserved glutamate residue is not relevant for the PatZ catalytic mechanism. Biophysical studies demonstrated that PatZ is a stable tetramer in solution and is transformed to its octameric form by autoacetylation. Moreover, this modification is reversed by the sirtuin CobB. Finally, an in silico PatZ tetramerization model based on hydrophobic and electrostatic interactions is proposed and validated by three-dimensional hydrodynamic analysis. These data reveal, for the first time, the structural regulation of an acetyltransferase by autoacetylation in a prokaryotic organism. 相似文献
94.
Lauriane Sollelis Mehdi Ghorbal Cameron Ross MacPherson Rafael Miyazawa Martins Nada Kuk Lucien Crobu Patrick Bastien Artur Scherf Jose‐Juan Lopez‐Rubio Yvon Sterkers 《Cellular microbiology》2015,17(10):1405-1412
Protozoan pathogens that cause leishmaniasis in humans are relatively refractory to genetic manipulation. In this work, we implemented the CRISPR‐Cas9 system in Leishmania parasites and demonstrated its efficient use for genome editing. The Cas9 endonuclease was expressed under the control of the Dihydrofolate Reductase‐Thymidylate Synthase (DHFR‐TS) promoter and the single guide RNA was produced under the control of the U6snRNA promoter and terminator. As a proof of concept, we chose to knockout a tandemly repeated gene family, the paraflagellar rod‐2 locus. We were able to obtain null mutants in a single round of transfection. In addition, we confirmed the absence of off‐target editions by whole genome sequencing of two independent clones. Our work demonstrates that CRISPR‐Cas9‐mediated gene knockout represents a major improvement in comparison with existing methods. Beyond gene knockout, this genome editing tool opens avenues for a multitude of functional studies to speed up research on leishmaniasis. 相似文献
95.
Discovery of a novel and conserved Plasmodium falciparum exported protein that is important for adhesion of PfEMP1 at the surface of infected erythrocytes
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Adéla Nacer Aurélie Claes Amy Roberts Christine Scheidig‐Benatar Hiroshi Sakamoto Mehdi Ghorbal Jose‐Juan Lopez‐Rubio Denise Mattei 《Cellular microbiology》2015,17(8):1205-1216
Plasmodium falciparum virulence is linked to its ability to sequester in post‐capillary venules in the human host. Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) is the main variant surface antigen implicated in this process. Complete loss of parasite adhesion is linked to a large subtelomeric deletion on chromosome 9 in a number of laboratory strains such as D10 and T9‐96. Similar to the cytoadherent reference line FCR3, D10 strain expresses PfEMP1 on the surface of parasitized erythrocytes, however without any detectable cytoadhesion. To investigate which of the deleted subtelomeric genes may be implicated in parasite adhesion, we selected 12 genes for D10 complementation studies that are predicted to code for proteins exported to the red blood cell. We identified a novel single copy gene (PF3D7_0936500) restricted to P. falciparum that restores adhesion to CD36, termed here virulence‐associated protein 1 (Pfvap1). Protein knockdown and gene knockout experiments confirmed a role of PfVAP1 in the adhesion process in FCR3 parasites. PfVAP1 is co‐exported with PfEMP1 into the host cell via vesicle‐like structures called Maurer's clefts. This study identifies a novel highly conserved parasite molecule that contributes to parasite virulence possibly by assisting PfEMP1 to establish functional adhesion at the host cell surface. 相似文献
96.
97.
Michael Dewaele Wim Martinet Noemí Rubio Tom Verfaillie Peter A. de Witte Jacques Piette Patrizia Agostinis 《Journal of cellular and molecular medicine》2011,15(6):1402-1414
Reactive oxygen species (ROS) concurrently instigate apoptosis and autophagy pathways, but the link between these processes remains unclear. Because cytotoxic ROS formation is exploited in anticancer therapy, such as in photodynamic therapy (PDT), a better understanding of the complex interplay between autophagy and apoptosis is urgently required. Previously, we reported that ROS generated by PDT with an endoplasmic reticulum (ER)-associated sensitizer leads to loss of ER-Ca2+ homeostasis, ER stress and apoptosis. Here we show that PDT prompted Akt-mTOR (mammalian target of rapamycin) pathway down-regulation and stimulated macroautophagy (MA) in cancer and normal cells. Overexpression of the antioxidant enzyme glutathione peroxidase-4 reversed mTOR down-regulation and blocked MA progression and apoptosis. Attenuating MA using Atg5 knockdown or 3-methyladenine, reduced clearance of oxidatively damaged proteins and increased apoptosis, thus revealing a cytoprotective role of MA in PDT. Paradoxically, genetic loss of MA improved clearance of oxidized proteins and reduced photokilling. We found that up-regulation of chaperone-mediated autophagy (CMA) in unstressed Atg5−/− cells compensated for MA loss and increased cellular resistance to PDT. CMA-deficient cells were significantly sensitized to photokilling but were protected against the ER stressor thapsigargin. These results disclose a stress-specific recruitment of autophagy pathways with cytoprotective function and unravel CMA as the dominant defence mechanism against PDT. 相似文献
98.
Rubio M Villain H Docagne F Roussel BD Ramos JA Vivien D Fernandez-Ruiz J Ali C 《PloS one》2011,6(8):e23690
Cessation of chronic ethanol consumption can increase the sensitivity of the brain to excitotoxic damages. Cannabinoids have been proposed as neuroprotectants in different models of neuronal injury, but their effect have never been investigated in a context of excitotoxicity after alcohol cessation. Here we examined the effects of the pharmacological activation/inhibition of the endocannabinoid system in an in vitro model of chronic ethanol exposure and withdrawal followed by an excitotoxic challenge. Ethanol withdrawal increased N-methyl-D-aspartate (NMDA)-evoked neuronal death, probably by altering the ratio between GluN2A and GluN2B NMDA receptor subunits. The stimulation of the endocannabinoid system with the cannabinoid agonist HU-210 decreased NMDA-induced neuronal death exclusively in ethanol-withdrawn neurons. This neuroprotection could be explained by a decrease in NMDA-stimulated calcium influx after the administration of HU-210, found exclusively in ethanol-withdrawn neurons. By contrast, the inhibition of the cannabinoid system with the CB1 receptor antagonist rimonabant (SR141716) during ethanol withdrawal increased death of ethanol-withdrawn neurons without any modification of NMDA-stimulated calcium influx. Moreover, chronic administration of rimonabant increased NMDA-stimulated toxicity not only in withdrawn neurons, but also in control neurons. In summary, we show for the first time that the stimulation of the endocannabinoid system is protective against the hyperexcitability developed during alcohol withdrawal. By contrast, the blockade of the endocannabinoid system is highly counterproductive during alcohol withdrawal. 相似文献
99.
100.
Plant establishment and growth on rocky outcrops in the Itatiaia massif (2400 m a.s.l.), southeastern Brazil, are limited by lack of soil and by freezing temperatures in winter nights. Mat‐forming species of different sizes and shapes on bare rock provide substrate for other plants to establish. The habitat preference of two geophytes, Stevia camporum (Asteraceae) and Alstroemeria foliosa (Alstroemeriaceae), was compared with regard to their association with the type of mat species and distinct rock topographies. The habitat preference of the mat species in regard to topography was also assessed. We found 1706 ramets of S. camporum and 1317 of A. foliosa in 253 vegetation islands ranging in size from 0.005 to 18.097 m2. Mat species in these islands were Fernseea itatiaiae, Vriesea itatiaiae (both Bromeliaceae), Pleurostima gounelleana (Velloziaceae) and/or Campylopus pilifer (Dicranaceae) and other mosses. Mat species were segregated by topography. Geophytes were similarly distributed across types of topography but showed negative interspecific association. S. camporum occurred mostly on the moss‐dominated islands, whereas A. foliosa was more common in P. gounelleana islands. We found geophyte co‐occurrence in larger vegetation islands containing both P. gounelleana and mosses as mat species. Thus, the effect of topography on geophyte segregation was indirect, since topography affects mat species distribution, and the geophyte preference for distinct mat species as substrate resulted in their segregation. 相似文献