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Mediator contributes to IgH locus VDJ rearrangements by promoting usage of most distal V segments

Cellular & Molecular Immunology 2020年第4期17卷 407-409页

作者： Iman Dalloul Zeinab Dalloul Sandrine Le Noir François Boyer Bernardo Reina-San-Martin Jeanne Cook-Moreau Michel Cogné UMR CNRS 7276 INSERM U1262Universite de LimogesLimoges 87025France INSERM U1258 UMR CNRS 7104Institut de Genetique et de Biologie Moleculaire et CellulaireUniversitéde StrasbourgIllkirch 67400France

INTRODUCTION The immunoglobulin heavy(IgH)chain locus includes a large cluster of V,D and J segments spanning thousands of kilobases in mammals and its expression implies long-range 3D-regulation of various gene modifications.1 V(D)J recombination occurs during early B-cell ontogeny.2 It is regulated by multiple transcription factors and cis-regulatory elements,and most notably by the intronic 5′Eμelement located in the JH to Cμintron.3 The intergenic control region 1(IGCR1)located between the VH and D gene clusters inhibits rearrangement of the most DH-proximal VH gene segments,while promoting usage of distal VH gene segments.4 Other downstream elements of the locus,flanking its 3′end might have more subtle roles:while the large 3′regulatory region(3′RR)shows strong internal synergies2 and behaves as a superenhancer in mature B *** deletion showed no influence on the VDJ repertoire and rather increased transcription of unrearranged V segments.3–5 Insulators binding CTCF farther downstream of the 3′RR were shown,however,to promote usage of distal VH segments.6 Altogether,there are strong indications that elements involved in long-distance regulation of transcription might also influence VDJ ***-frame splicing of VDJ exons is also needed for full Ig expression.7 Given the functional importance of gene architecture and long-range interactions for the IgH locus physiology,we explored the dependence of the VDJ repertoire on Med1,a known actor of long-range enhancer interactions.

关键词： B cells mediator VDJ repertoire

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NTR/NRX Define a New Thioredoxin System in the Nucleus of Arabidopsis thaliana Cells

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Molecular Plant 2014年第1期7卷 30-44页

作者： Corinne Marchal Valerie Delorme-Hinoux Laetitia Bariat Wafi Siala Christophe Belin Julio Saez-Vasquez Christophe Riondet Jean-Philippe Reichheld Universite Perpignan Via Domitia CNRS Laboratoire Genome et Developpement des Plantes UMR 5096 58 Avenue Paul Alduy-Bat T F-66860 Perpignan France Pharmacologie cellulaire et moleculaire Louvain Drug Research Institute Universite catholique de Louvain Brussels Belgium

Thioredoxins （TRX） are key components of cellular redox balance, regulating many target proteins through thiol/disulfide exchange reactions. In higher plants, TRX constitute a complex multigenic family whose members have been found in almost all cellular compartments. Although chloroplastic and cytosolic TRX systems have been largely studied, the presence of a nuclear TRX system has been elusive for a long time. Nucleoredoxins （NRX） are potential nuclear TRX found in most eukaryotic organisms. In contrast to mammals, which harbor a unique NRX, angiosperms gen- erally possess multiple NRX organized in three subfamilies. Here, we show thatArabidopsis thaliana has two NRXgenes （AtNRX1 and AtNRX2）, respectively, belonging to subgroups I and III. While NRX1 harbors typical TRX active sites （WCG/ PPC）, NRX2 has atypical active sites （WCRPC and WCPPF）. Nevertheless, both NRXl and NRX2 have disulfide reduction capacities, although NRX1 alone can be reduced by the thioredoxin reductase NTRA. We also show that both NRX1 and NRX2 have a dual nuclear/cytosolic localization. Interestingly, we found that NTRA, previously identified as a cytosolic protein, is also partially localized in the nucleus, suggesting that a complete TRX system is functional in the nucleus. We show that NRX1 is mainly found as a dimer in vivo. nrxl and nrx2 knockout mutant plants exhibit no phenotypic perturbations under standard growth conditions. However, the nrxl mutant shows a reduced pollen fertility phenotype, suggesting a specific role of NRX1 at the haploid phase.

关键词： thioredoxin nucleoredoxin nucleus Arabidopsis.

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Mutations of the aurora kinase C gene causing macrozoospermia are the most frequent genetic cause of male infertility in Algerian men

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Asian Journal of Andrology 2015年第1期17卷 68-73,I0008页

作者： Leyla Ounis Abdelali Zoghmar Charles Coutton Leila Rouabah Maroua Hachemil Delphine Martinez Guillaume Martinez Ines Bellil Douadi Khelifi Christophe Arnoult Julien Faure Sebti Benbouhedja Abdelkader Rouabah Pierre F Ray Laboratoire de Biologie moleculaire et cellulaire Faculte des Sciences de la Nature et de la Vie Universite Constantine I Algerie Clinique Ibn Rochd Centre de Medecine de la Reproduction Constantine Algerie Universite Grenoble Alpes Grenoble France Equipe Genetique Epigenetique et Therapies de I'lnfertilitE CNRS AGIM Grenoble France Laboratoire de GEnetique Chromosomique CHU Grenoble Grenoble France Laboratoire de Biochimie et Genetique moleculaire CHU Grenoble Grenoble France Equipe Muscle et Pathologies INSERM U836 Grenoble Institut des Neurosciences Grenoble France.

Klinefelter syndrome and Y-chromosomal microdeletion analyses were once the only two genetic tests offered to infertile men. Analyses of aurora kinase C （AURKCj and DPY19L2 are now recommended for patients presenting macrozoospermia and globozoospermia, respectively, two rare forms of teratozoospermia particularly frequent among North African men. We carried out genetic analyses on Algerian patients, to evaluate the prevalence of these syndromes in this population and to compare it with the expected frequency of Klinefelter syndrome and Y-microdeletions. We carried out a retrospective study on 599 consecutive patients consulting for couple infertility at the assisted reproduction unit of the Ibn Rochd Clinique, Constantine, Algeria. Abnormal sperm parameters were observed in 404 men. Fourteen and seven men had typical macrozoospermia and globozoospermia profiles, respectively. Molecular diagnosis was carried out for these patients, for the AURKC and DPY19L2 genes. Eleven men with macrozoospermia had a homozygous AURKC mutation （79%）, corresponding to 2.7% of all patients with abnormal spermograms. All the men with globozoospermia studied （n = 5）, corresponding to 1.2% of all infertile men, presented a homozygous DPY19L2deletion. By comparison, we would expect 1.6% of the patients in this cohort to have Klinefelter syndrome and 0.23% to have Y-microdeletion. Our findings thus indicate that AURKCmutations are more frequent than Klinefelter syndrome and constitute the leading genetic cause of infertility in North African men. Furthermore, we estimate that AURKCand DPY19L2 molecular defects are 10 and 5 times more frequent, respectively, than Y-microdeletions.

关键词： aurora kinase C DPY19L2 globozoospermia intracytoplasmic sperm injection infertility macrozoospermia

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High-Resolution Crystal Structure and Redox Properties of Chloroplastic Triosephosphate Isomerase from Chlamydomonas reinhardtii

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Molecular Plant 2014年第1期7卷 101-120页

作者： Mirko Zaffagnini Laure Michelet Chiara Sciabolini Nastasia Di Giacinto Samuel Morisse Christophe H. Marchand Paolo Trost Simona Fermani Stephane D Lemaire Laboratoire de Biologie moleculaire et cellulaire des Eucaryotes FRE3354 Centre National de la Recherche Scientifique Universite Pierre et Marie Curie Institut de Biologie Physico-Chimique 13 rue Pierre et Marie Curie 75005 Paris France Laboratory of Plant Redox Biology Department of Pharmacy and Biotechnology (FaBiT) University of Bologna via Irnerio 42 40126 Bologna Italy Department of Chemistry 'G. Ciamician' University of Bologna via Selmi 2 40126 Bologna Italy

Triosephosphate isomerase （TPI） catalyzes the interconversion of glyceraldehyde-3-phosphate to dihydroxyacetone phosphate. Photosynthetic organisms generally contain two isoforms of TPI located in both cytoplasm and chloroplasts. While the cytoplasmic TPI is involved in the glycolysis, the chloroplastic isoform participates in the Calvin-Benson cycle, a key photosynthetic process responsible for carbon fixation. Compared with its cytoplasmic counterpart, the functional features of chloroplastic TPI have been poorly investigated and its three-dimensional structure has not been solved. Recently, several studies proposed TPI as a potential target of different redox modifications including dithiol/disulfide interchanges, glutathionylation, and nitrosylation. However, neither the effects on protein activity nor the molecular mechanisms underlying these redox modifications have been investigated. Here, we have produced recombinantly and purified TPI from the unicellular green alga Chlamydomonas reinhardtii （Cr）. The biochemical properties of the enzyme were delineated and its crystallographic structure was determined at a resolution of 1.1 A. CrTPI is a homodimer with subunits containing the typical （β/α）8-barrel fold. Although no evidence for TRX regulation was obtained, CrTPI was found to undergo glutathionylation by oxidized glutathione and trans-nitrosylation by nitrosoglutathione, confirming its sensitivity to multiple redox modifications.

关键词： triosephosphate isomerase three-dimensional structure TIM-barrel thiol-based redox regulation transnitrosylation.

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Effect of Mitochondrial Dysfunction on Carbon Metabolism and Gene Expression in Flower Tissues of Arabidopsis thaliana

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Molecular Plant 2011年第1期4卷 127-143页

作者： Maria V. Busi Maria E. Gomez-Lobato Sebastian P. Rius Valeria R. Turowski Paula Casati Eduardo J. Zabaleta Diego F. Gomez-Casati Alejandro Araya Instituto de Investigaciones Biotecnologicas-lnstituto Tecnologico de ChascomOs (IIB-INTECH) CONICet/UNSAM Chascomus Argentina Centro de Estudios Fotosinteticos y Bioquimicos (CEFOBI) Universidad Nacional de Rosario Argentina Instituto de Investigaciones Biologicas liB CONICET-Universidad Nacional de Mar del Plata Argentina microbiologie cellulaire et moleculaire et pathogenicite UMR 5234 Centre National de la Recherche Scientifique and Universite Victor Segalen-Bordeaux 2 Bordeaux France

We characterized the transcriptomic response of transgenic plants carrying a mitochondrial dysfunction induced by the expression of the unedited form of the ATP synthase subunit 9. The u-ATP9 transgene driven by A9 and APetALA3 promoters induce mitochondrial dysfunction revealed by a decrease in both oxygen uptake and adenine nucleo- tides （ATP, ADP） levels without changes in the ATIP/ADP ratio. Furthermore, we measured an increase in ROS accumulation and a decrease in glutathione and ascorbate levels with a concomitant oxidative stress response. The transcriptome analysis of young Arabidopsis flowers, validated by qRT-PCR and enzymatic or functional tests, showed dramatic changes in u-ATP9 plants. Both lines display a modification in the expression of various genes involved in carbon, lipid, and cell wall metabolism, suggesting that an important metabolic readjustment occurs in plants with a mitochondrial dysfunction. Interestingly, transcript levels involved in mitochondrial respiration, protein synthesis, and degradation are affected. More- over, the levels of several mRNAs encoding for transcription factors and DNA binding proteins were also changed. Some of them are involved in stress and hormone responses, suggesting that several signaling pathways overlap. Indeed, the tran- scriptome data revealed that the mitochondrial dysfunction dramatically alters the expression of genes involved in signaling pathways, including those related to ethylene, absicic acid, and auxin signal transduction. Our data suggest that the mitochondrial dysfunction model used in this report may be useful to uncover the retrograde signaling mechanism between the nucleus and mitochondria in plant cells.

关键词： Electron transport transcriptome analysis Arabidopsis.

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Evaluation of in vitro antioxidant and apoptotic activities of Cyperus rotundus

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Asian Pacific Journal of Tropical Medicine 2014年第2期7卷 105-112页

作者： Kilani-Jaziri Soumaya Ghedira Zied Nasr Nouha Krifa Mounira Ghedira Kamel Franca Dijoux Marie Genviève Ghedira Chekir Leila Unité de Substances Naturelles Bioactives et Biotechnologies UR12ES12.Universite de Monastic Faculte de Pharmacie de Monastir Laboratoire de Biologie moleculaire et cellulaire Faculté de Medecine Dentaire de MonastirUniversite de MonastirRue Aricenne5019 MonastirTunisie Laboratoire de Botanique.Pharmacognosie et Phytothérapie UMR CNRS 5557 Ecologie Microbienne Institut des Sciences Pharmaceutiques et Biologiques-Faculte de Pharmacie Université Claude Bernard Btiment Netien-8 Avenue Rockefeller-69373 Lyon cedex 08France

Objective:To evaluate in vitro antioxidant and apoptotic activities of Crperus rotundas(***).Methods:The phytochemical study and the antioxidant activities of both methanol and aqueous extracts from *** aerial part were *** addition,these extracts were also investigated for their cytotoxic and apoptotic *** major compound of the methanol extract was *** metlianol and aqueous extracts(300,150,and 50μg/mL)were evaluated for their antioxidant activity by the xanthine/xanthine oxidase assay ***,16,8,and 4 mg/mL of each extract were tested to investigate their *** scavenging *** extract(800,*** 200μg/mL)and melhunol extract(350,175,and 88μg/mL)were tested against lipid peroxidation,induced by 75μM H_2O_2,The cytotoxicity(by MTT assay)and cell DNA fragmentation of both extracts were evaluated Inwards K562 and L1210 cell *** major compound was obtained from the butanol fraction of methanol extract and its structure was determined by KMN spectroscopic ***:The methanol and aqueous extracts showed respectively,88%and 19%inhibition of xanthine oxidase *** same extracts inhibited lipid peroxidation by 61.5%and 42.0%.*** extracts inhibited *** by 27.1%and 25.3%,*** methanol cxtract induced DNA *** was determined as the major compound isolated from the butanol fraction of metlianol ***:It appears that *** extracts exhibit a potential use as a natural antioxidant and an apoptosis inducer.

关键词： Polyphenols Orientin Xanthine/Xanthine oxidase assay Lipid peroxidation assay Deoxyribose assay DNA fragmentation

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A LIM Domain Protein from Tobacco Involved in Actin-Bundling and Histone Gene Transcription

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Molecular Plant 2013年第2期6卷 483-502页

作者： Danièle Moes Sabrina Gatti CEline Hoffmann Monika Dieterle Flora Moreau Katrin Neumann Marc Schumacher Marc Diederich Erwin Grill Wen-Hui Shen André Steinmetz Cldment Thomas Centre de Recherche Public-SantE 84 Val Fleuri L-1526 Luxembourg Luxembourg Laboratoire de Biologie moleculaire et cellulaire du Cancer HSpital Kirchberg 9 rue Edward Steichen L-2540 Luxembourg Luxembourg Lehrstuhl for Botanik Technische Universitat Munchen EmiI-Ramann-StraBe 4 D-85354 Freising Germany Jnstitut de Biologie moleculaire des Plantes du CNRS 12 rue du Général Zimmer F-67084 Strasbourg Ceédex France

The two LIM domain-containing proteins from plants （LIMs） typically exhibit a dual cytoplasmic-nuclear dis-tribution, suggesting that, in addition to their previously described roles in actin cytoskeleton organization, they partici-pate in nuclear processes. Using a south-western blot-based screen aimed at identifying factors that bind to plant histone gene promoters, we isolated a positive clone containing the tobacco LIM protein WLIM2 （NtWLIM2） cDNA. Using both green fluorescent protein （GFP） fusion-and immunology-based strategies, we provide clear evidence that NtWLIM2 local-izes to the actin cytoskeleton, the nucleus, and the nucleolus. Interestingly, the disruption of the actin cytoskeleton by latrunculin B significantly increases NtWLIM2 nuclear fraction, pinpointing a possible novel cytoskeletal-nuclear crosstalk. Biochemical and electron microscopy experiments reveal the ability of NtWLIM2 to directly bind to actin filaments and to crosslink the latter into thick actin bundles. Electrophoretic mobility shift assays show that NtWLIM2 specifically binds to the conserved octameric cis-elements （Oct） of the Arabidopsis histone H4A748 gene promoter and that this binding largely relies on both LIM domains. Importantly, reporter-based experiments conducted in Arabidopsis and tobacco proto-plasts confirm the ability of NtWLIM2 to bind to and activate the H4A748 gene promoter in live cells. Expression studies indicate the constitutive presence of NtWLIM2 mRNA and NtWLIM2 protein during tobacco BY-2 cell proliferation and cell cycle progression, suggesting a role of NtWLIM2 in the activation of basal histone gene expression. Interestingly, both live cell and in vitro data support NtWLIM2 di/oligomerization. We propose that NtWLIM2 functions as an actin-stabilizing protein, which, upon cytoskeleton remodeling, shuttles to the nucleus in order to modify gene expression.

关键词： actin BY-2 cytoskeleton DNA-binding histone genes LIM Nicotiana tabacum promoter regulation trans-acting factors.

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Improved maize growth in condition controlled by PGPR inoculation on ferruginous soil in central Benin

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Food and Nutrition Sciences 2019年第12期10卷 1433-1451页

作者： Olarewadjou Amogou Nadege Adouke Agbodjato Gustave Dagbenonbakin Pacome Agossou Noumavo Haziz Sina Assogba Abado Sylvestre Marcel Yevedo Adoko Martial Nounagnon Romain Glele Kakai Adolphe Adjanohoun Lamine Baba-Moussa Laboratoire de Biologie et de Typage moleculaire en microbiologie Departement de Biochimie et de Biologie CellulaireFaculte des Sciences et TechniquesUniversite d’Abomey-CalaviCotonouBenin Centre de Recherches Agricoles d’Agonkanmey Institut National des Recherches Agricoles du BeninCotonouBenin Laboratoire de microbiologie et des Technologies Alimentaires Departement de Biologie VegetaleFaculte des Sciences et TechniquesUniversite d’Abomey-CalaviCotonouBenin Laboratoire de Biomathematique et d’Estimation Forestiere Faculty of Agronomic SciencesUniversity of Abomey-CalaviCotonouBenin

Abstract The use of microbial technologies in agriculture is rapidly expanding with the discovery of new bacterial strains effective in improving plant growth. In this study, we tested and highlighted the efficacy of PGPR (Plant Growth Promoting Rhizobacteria) alone or in a consortium on maize growth. For this purpose, a greenhouse experiment was carried out in pots containing sterilized ferruginous soil for 30 days. The corn seeds of the EVDT 97 SRT C1 variety were inoculated with bacterial suspensions of concentration 108 UFC/ml. The experimental device was a random block of 16 three-repeat treatments. The incidence of PGPR inoculated strains is assessed on the biomass growth and yield parameters of maize. At the end of the trial, the results showed that inoculation stimulated plant growth and development and resulted in a significant increase in the height, diameter at the collar, leaf surface and dry weight of aerial biomass. 20.15%, 21%, 32.77% and 37.73% respectively compared to controls, especially in corn plants inoculated with B. thurengiensis;*** ***- *** et Pseudomonas cichorii- Pseudomonas. putida- Pseudomonas syringae. These results show the potential of using these rhizobacteria as biological inoculants to improve maize productivity in Benin.

关键词： PGPR PGPR Inoculation Growth Biomass Ferruginous Soil

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Influence of Isolated PGPR Rhizobacteria in Central and Northern Benin on Maize Germination and Greenhouse Growth

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American Journal of Plant Sciences 2018年第13期9卷 2775-2793页

作者： Olaréwadjou Amogou Gustave Dagbénonbakin Nadege Adouke Agbodjato Pacome Agossou Noumavo Hafiz Adio Salami Salako Valere Aguegue Mevognon Ricardos Assogba Abado Sylvestre Koda Fousseni Abdel Djihal Adolphe Adjanohoun Lamine Baba-Moussa Departement de Biochimie et de Bio-logie cellulaire Faculte des Sciences et Techniques(FAST)Laboratoire de Biologie et de Typage Moleculaire en MicrobiologieUniversite d’Abomey-Calavi(UAC)CotonouBénin Centre de Recherches Agricoles d’Agonkanmey Institut National des Recherches Agricoles du BéninCotonouBénin Département de Biologie Végétale Laboratoire de Microbiologie et des Technologies AlimentairesCotonouBénin Faculte des Science Agronomique(FSA) Laboratoire de Biomathématique et d’Estimation ForestièreUniversite d’Abomey-Calavi(UAC)CotonouBénin Centre de Recherches Agricoles Sud Institut National des Recherches Agricoles du BéninAttogonBénin

The objective of the study was to evaluate the effects of nine plant growth-promoting rhizobacteria (PGPR) alone or in combination on maize seed germination and seedling growth under laboratory and greenhouse conditions. The germination and growth tests were carried out in square petri dishes and pots. Maize seeds were inoculated with suspension of 108 CFU/ml of rhizobacteria. The experimental device was a random block of 16 treatments with four repetitions. Germination test results showed that seeds inoculated with PGPR including the control induced good germination in the range of 93.75% to 100%. The vigor index and root length of the seeds treated with Bacillus panthothenicus were significantly improved by 76.64% and 58.86%, respectively, while the maximal lengths of the seedlings were obtained with Pseudomonas cichorii with an increase of 118.95%. In greenhouse experience, data demonstrated that Serratia marcescens better improved the leaf area, height and underground biomass, respectively by 58.83%, 108.43%, and 59.16% as compared to the control. The highest fresh aerial biomass and air dry matter was obtained with plants treated only with Pseudomonas putida. These results show the potential to use such rhizobacteria as biofertilizers to improve maize productivity in Benin.

关键词： PGPR Biofertilization Germination Growth Soil Ferruginous

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Two Highly Variable Vpr<sub>84</sub>and Vpr<sub>85</sub>Residues within the HIV-1-Vpr C-Terminal Protein Transduction Domain Control Transductionnal Activity and Define a Clade Specific Polymorphism

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World Journal of AIDS 2014年第2期4卷 148-155页

作者： Jean-Herve Colle Thiery Rose Christine Rouzioux Alphonse Garcia EA 3620 Universite Paris Descartes AP-HP Laboratoire de Virologie Hopital Necker Paris France Laboratoire E3 Phosphatases Unite Signalisation Moleculaire et Activation Cellulaire Institut Pasteur Paris France Unite d’Immunogenetique cellulaire Institut Pasteur Paris France Unite de Biologie des Populations Lymphocytaires Institut Pasteur Paris France

The virally encoded HIV-1 viral protein R (VPR) is a multifunctional factor that is required for induced HIV-1 pathogenesis. VPR is also a cell-penetrating protein found in biological fluids from HIV-1 infected individuals. In this regard, we previously published that the C-terminal VPR77-92 sequence from HIV-1 89.6, but not from pNL4.3 strain, is a new pro-apoptotic and protein transduction domain (PTD). Here we report on a sequence analysis of VPR77-92 domain using the Los Alamos HIV-1 sequence database. The analysis showed that the two residues of the domain VPR84 and VPR85 are highly variable and differently biased in HIV-1 clade B and HIV-1 clade C. Furthermore, when Jurkat lymphoblastoid cells or PBMC were incubated with chemically synthesized peptides containing distinct VPR77-92 C-terminal sequences from clades B or C, we found that a clade-dependent polymorphism in VPR84 and VPR85 residues controlled the transducing activity of the C-terminal HIV-1 VPR77-92 domain. Together our data indicate that clade-dependent polymorphism in the VPR84 and VPR85?residues defines the transducing properties mediated by the C-terminal domain of HIV-1 VPR. Identification of this VPR polymorphism suggests new approaches to understand the HIV-1 biology and/or pathogenesis.

关键词： VPR HIV-1 Subtypes Polymorphism Transduction

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