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See detailClonage, caractérisation et étude de la régulation transcriptionnelle du gène Aox1 encodant l'oxydase alternative chez Chlamydomonas reinhardtii
Baurain, Denis ULg

Doctoral thesis (2003)

Au sein de la membrane interne des mitochondries, quatre complexes multiprotéiques sont impliqués dans le transfert des électrons depuis les équivalents réducteurs jusqu'à l'oxygène moléculaire. L'énergie ... [more ▼]

Au sein de la membrane interne des mitochondries, quatre complexes multiprotéiques sont impliqués dans le transfert des électrons depuis les équivalents réducteurs jusqu'à l'oxygène moléculaire. L'énergie associée à ce transport au travers des complexes I, III et IV est couplée à la synthèse d'ATP par l'intermédiaire d'un gradient de protons. Chez les plantes supérieures, de nombreux champignons et quelques protistes, une seconde voie de transfert diverge de la voie principale au niveau du pool d'ubiquinone, celui-ci étant alors oxydé directement par l'oxygène moléculaire. Lorsque les électrons empruntent cette voie alternative, deux sites d'éjection de protons sont court-circuités et l'énergie produite est dissipée sous forme de chaleur. Cette réaction est catalysée par une enzyme unique, l'oxydase alternative (AOX), souvent encodée par une petite famille multigénique chez les plantes supérieures. Une activité accrue de la voie alternative est observée suite à divers stimuli développementaux et environnementaux, en particulier en conditions de stress. Cette augmentation d'activité résulte d'une activation transcriptionnelle du gène Aox et/ou de modifications post-traductionnelles de la protéine mature. L'AOX de l'algue verte unicellulaire Chlamydomonas reinhardtii est encodée par deux gènes différents, Aox1 et Aox2, le premier étant beaucoup plus transcrit que le second. Les cDNAs Aox1 et Aox2, de même que la séquence génomique Aox2, ont été isolés et caractérisés dans notre laboratoire. Dans un premier temps, nous avons entrepris le clonage et la caractérisation de la séquence génomique Aox1, ce qui nous a permis de comparer sa structure avec celle de son homologue Aox2. Ensuite, afin d'étudier sa régulation transcriptionnelle, nous avons fusionné un segment de 1,4 kb contenant la région promotrice Aox1 à la région codante du gène (Ars) de l'arylsulfatase et mesuré les activités ARS dans des transformants porteurs de la construction chimérique. Nous avons ainsi montré que le promoteur Aox1 est insensible à la plupart des inducteurs classiques de l'AOX, parmi lesquels des agents de stress, des inhibiteurs respiratoires et des métabolites. En revanche, l'expression du gène Aox1 répond à la nature de la source d'azote, sa transcription étant réprimée par l'ammonium et stimulée par le nitrate. De plus, en milieu contenant du nitrate, l'inactivation de la nitrate réductase (première enzyme de la voie d'assimilation du nitrate) conduit à une expression du gène Aox1 encore plus importante. Nous avons en outre observé que cette stimulation par le nitrate se répercute aux niveaux protéique et respiratoire. Une étude de délétion de la région promotrice Aox1 indique qu'un segment court (de −253 à +59 par rapport à l'origine de transcription) est suffisant pour assurer la transcription et la régulation du gène, mais que son expression maximale requiert également des éléments distaux. Aucun motif nucléotidique susceptible d'intervenir dans l'expression du gène Aox1 n'a été identifié à l'issue d'une analyse bioinformatique du promoteur. L'effet de la nature de la source d'azote sur l'expression de l'AOX est interprété sous l'angle d'une optimisation de la synthèse d'ATP mitochondrial sans modification de l'activité respiratoire, en relation avec un possible accroissement de la production d'ATP photosynthétique lorsque le nitrate est utilisé comme source d'azote. [less ▲]

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See detailClonal chromosome aberrations in Philadelphia-negative cells from chronic myelocytic leukemia patients treated with imatinib mesylate: report of two cases.
Herens, Christian ULg; Baron, Frédéric ULg; Croisiau, Christiane et al

in Cancer Genetics & Cytogenetics (2003), 147(1), 78-80

Imatinib mesylate (tested as STI571), an abl kinase inhibitor, induces sustained, complete hematologic and cytogenetic responses in chronic myelocytic leukemia (CML) patients; however, emergence of clonal ... [more ▼]

Imatinib mesylate (tested as STI571), an abl kinase inhibitor, induces sustained, complete hematologic and cytogenetic responses in chronic myelocytic leukemia (CML) patients; however, emergence of clonal chromosomal aberrations in Philadelphia-negative (Ph-) cells during treatment has been reported. We describe two CML patients in chronic phase who presented with complete cytogenetic responses during imatinib mesylate therapy but developed new clonal chromosomal rearrangements in Ph- cells. The first patient presented with a duplication of chromosome 1, dup(1)(q21q42), and the second showed two new clonal aberrations consisting of inv(1)(q12q32) and del(7)(q22) in the same clone. [less ▲]

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See detailCloned Human Polyomavirus JC Can Transform Human Amnion Cells
Howley, P. M.; Rentier-Delrue, Françoise ULg; Heilman, C. A. et al

in Journal of Virology (1980), 36

The genome of the human polyomavirus JC (Mad-1 strain) was molecularly cloned in Escherichia coli by using the plasmid vector pBR322. Recombinant DNA molecules were constructed with the entire JC genome ... [more ▼]

The genome of the human polyomavirus JC (Mad-1 strain) was molecularly cloned in Escherichia coli by using the plasmid vector pBR322. Recombinant DNA molecules were constructed with the entire JC genome inserted either at its unique EcoRI site at 0.0 map units or at its unique BamHI site at 0.51 map units. Viral DNA from each of these recombinant plasmids was capable of transforming human amnion cells, and cell lines established from transformed foci were positive for JC tumor antigen as assayed by indirect immunofluorescence. [less ▲]

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See detailClonidine and ketanserin both are effective treatment for postanesthetic shivering.
Joris, Jean ULg; Banache, Maryse; Bonnet, Francis et al

in Anesthesiology (1993), 79(3), 532-9

BACKGROUND: Although meperidine is an effective treatment of postanesthetic shivering, its mechanism of action remains unknown. Investigation of other drugs might help clarify the mechanisms by which ... [more ▼]

BACKGROUND: Although meperidine is an effective treatment of postanesthetic shivering, its mechanism of action remains unknown. Investigation of other drugs might help clarify the mechanisms by which shivering can be controlled. Accordingly, we investigated the efficacy of clonidine, an alpha 2-adrenergic agonist, and ketanserin, a 5-hydroxytryptamine antagonist, in treating postanesthetic shivering. METHODS: First, 54 patients shivering after general anesthesia were allocated randomly to receive an intravenous bolus of saline, 150 micrograms clonidine, or 10 mg ketanserin. A second study explored the dose-dependence of clonidine. Forty shivering patients were given saline or clonidine, 37.5, 75, or 150 micrograms. RESULTS: The duration of shivering was significantly shorter in those given clonidine (2.1 +/- 0.9 min) than in the other two groups and shorter in the ketanserin group (4.3 +/- 0.9 min) than in the saline group (12.0 +/- 1.6 min). Clonidine and ketanserin significantly decreased systolic arterial blood pressure when compared to saline. Core rewarming was significantly slower in the clonidine group. In the second study, 37.5 micrograms clonidine was no more effective than saline. Two minutes after treatment, 150 micrograms obliterated shivering in all patients. Five minutes after treatment, all patients given 75 micrograms had stopped shivering. Systolic arterial pressure and heart rate decreased significantly in patients given 75 and 150 micrograms clonidine. CONCLUSIONS: Clonidine (150 micrograms) and ketanserin (10 mg) both are effective treatment for postanesthetic shivering. The effect of clonidine on shivering is dose-dependent: whereas 37.5 micrograms had no effect, 75 micrograms clonidine stopped shivering within 5 min. [less ▲]

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See detailThe clonidine test in posttraumatic stress disorder.
Hansenne, Michel ULg; Pitchot, William ULg; Ansseau, Marc ULg

in American Journal of Psychiatry (The) (1991), 148(6), 810-1

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See detailCloning and amplified expression in Streptomyces lividans of a gene encoding extracellular β-lactamase from Streptomyces albus G
Dehottay, Philippe; Dusart, Jean; Duez, Colette ULg et al

in Gene (1986), 42(1), 31-36

A 4.9-kb DNA fragment containing the bla gene for the extracellular beta-lactamase (BLA) of Streptomyces albus G was cloned in Streptomyces lividans using the conjugative, low-copy-number plasmid pIJ61 as ... [more ▼]

A 4.9-kb DNA fragment containing the bla gene for the extracellular beta-lactamase (BLA) of Streptomyces albus G was cloned in Streptomyces lividans using the conjugative, low-copy-number plasmid pIJ61 as vector. No expression of bla was observed when this DNA fragment was introduced into Escherichia coli HB101 on a plasmid vector. A 1.5-kb PstI-SstI fragment containing the bla gene was cloned in S. lividans on the nonconjugative, high-copy-number plasmid pIJ702. A tenfold higher yield of BLA was obtained from S. lividans carrying this plasmid than from S. albus G grown under optimal production conditions. The BLA from the clone reacts with beta-iodopenicillanate according to a branched pathway which is characteristic of the original S. albus G BLA enzyme. [less ▲]

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See detailCloning and amplified expression in Streptomyces lividans of the gene encoding the extracellular β-lactamase from Streptomyces cacaoi
Lenzini, Mauro V; Nojima, Shiego; Dusart, Jean et al

in Journal of General Microbiology (1987), 133(10), 2915-2920

A 19 kb SphI DNA fragment containing the gene for the extracellular active-site serine beta-lactamase of Streptomyces cacaoi KCC-SO352 was cloned in Streptomyces lividans TK24 using the high-copy-number ... [more ▼]

A 19 kb SphI DNA fragment containing the gene for the extracellular active-site serine beta-lactamase of Streptomyces cacaoi KCC-SO352 was cloned in Streptomyces lividans TK24 using the high-copy-number plasmid pIJ702 as vector. A 30-fold higher yield of beta-lactamase was obtained from S. lividans strain ML1, carrying the recombinant plasmid pDML51, than from S. cacaoi grown under optimal production conditions. In all respects (molecular mass, isoelectric point, kinetics of inhibition by beta-iodopenicillanate) the overproduced S. lividans ML1 beta-lactamase was identical to the original S. cacaoi enzyme. A considerable reduction of beta-lactamase production was caused by elimination of a 12.8 kb portion of the 19 kb DNA fragment by cleavage at an internal SphI site located more than 3 kb upstream of the beta-lactamase structural gene. The beta-lactamase gene was located within a 1.8 NcoI-BclI fragment but when this fragment was cloned in S. lividans pIJ702, the resulting strain produced hardly any more beta-lactamase than the original S. cacaoi. [less ▲]

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See detailCloning and amplified expression in Streptomyces lividans of the gene encoding the extracellular β-lactamase of Actinomadura R39
Fraipont, Claudine ULg; Duez, Colette ULg; Matagne, André ULg et al

in Biochemical Journal (1989), 262(3), 849-854

By using the promoter-probe plasmid pIJ424, genomic DNA fragments of Actinomadura R39 were shown to have promoter activity in Streptomyces lividans. The same 100-200-copy-number plasmid was used to clone ... [more ▼]

By using the promoter-probe plasmid pIJ424, genomic DNA fragments of Actinomadura R39 were shown to have promoter activity in Streptomyces lividans. The same 100-200-copy-number plasmid was used to clone in S. lividans TK24, the gene that encodes the Actinomadura R39, β-lactamase. Gene cloning resulted in an amplified expression of the , β lactamase when compared with the amounts of enzyme produced by the original strain (1 mg versus 0.008 mg . litre of culture-1). [less ▲]

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See detailCloning and characterisation of the primary structure of the sheep lymphocyte function-associated antigen-1 alpha subunit
Fett, Thomas ULg; Zecchinon, Laurent ULg; Baise, Etienne ULg et al

in Molecular Immunology (2005), 42(12), 1503-1508

The leukocyte integrins play a critical role in a number of cellular adhesive interactions during the immune response. The ovine cDNA encoding CD1 1a, the predominant a subunit of the beta(2)-integrin ... [more ▼]

The leukocyte integrins play a critical role in a number of cellular adhesive interactions during the immune response. The ovine cDNA encoding CD1 1a, the predominant a subunit of the beta(2)-integrin family, was sequenced and compared with the human, bovine and murine sequences. Despite some focal differences, it shares all the main characteristics of its known mammalian homologues. Along with the ovine CD18-encoding cDNA, which is available for a few months, the sequence data provided here will allow the Ovis aries beta(2)-integrin CD1 1a/CD18 (LEA-1, alpha(L)beta 2) expression in vitro as a tool to examine the specificities of inflammation in the ovine species. (c) 2005 Elsevier Ltd. All rights reserved. [less ▲]

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See detailCloning and characterization of a cDNA encoding the beta-subunit of the bovine insulin-like growth factor-1 receptor.
Sneyers, M.; Kettmann, Richard ULg; Massart, Serge et al

in DNA Sequence : The Journal of DNA Sequencing & Mapping (1991), 1(6),

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See detailCloning and characterization of a cDNA encoding the bovine insulin-like growth factor binding protein 1 (bIGFBP-1).
Sneyers, M.; Kettmann, Richard ULg; Massart, Serge et al

in DNA Sequence : The Journal of DNA Sequencing & Mapping (1991), 1(6),

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See detailCloning and characterization of a cDNA encoding the bovine insulin-like growth factor binding protein I (bIGF-I)
Sneyers, Myriam; Kettmann, Richard ULg; Massart, Serge et al

in III Inter Symp on Molecular and Cell Biol of Insulin and IGF's (1990)

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See detailCloning and characterization of a cDNA encoding the ß-subunit of the bovine insulin-like growth factor-I receptor.
Sneyers, Myriam; Kettmann, Richard ULg; Massart, Serge et al

Poster (1991)

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See detailCloning and characterization of ADAMTS-14, a novel ADAMTS displaying high homology with ADAMTS-2 and ADAMTS-3.
Colige, Alain ULg; Vandenberghe, Isabel; Thiry, Marc ULg et al

in Journal of Biological Chemistry (2002), 277(8), 5756-66

The processing of amino- and carboxyl-propeptides of fibrillar collagens is required to generate collagen monomers that correctly assemble into fibrils. Mutations in the ADAMTS2 gene, the ... [more ▼]

The processing of amino- and carboxyl-propeptides of fibrillar collagens is required to generate collagen monomers that correctly assemble into fibrils. Mutations in the ADAMTS2 gene, the aminopropeptidase of procollagen I and II, result in the accumulation of non-fully processed type I procollagen, causing human Ehlers-Danlos syndrome type VIIC and animal dermatosparaxis. In this study, we show that the aminopropeptide of type I procollagen can be cleaved in vivo in absence of ADAMTS-2 activity and that this processing is performed at the cleavage site for ADAMTS-2. In an attempt to identify the enzyme responsible for this alternative aminoprocollagen peptidase activity, we have cloned the cDNA and determined the primary structure of human and mouse ADAMTS-14, a novel ADAMTS displaying striking homologies with ADAMTS-2 and -3. The structure of the human gene, which maps to 10q21.3, and the mechanisms of generation of the various transcripts are described. The existence of two sites of initiation of transcription, in two different promoter contexts, suggests that transcripts resulting from these two sites can be differently regulated. The tissue distribution of ADAMTS-14, the regulation of the gene expression by various cytokines and the activity of the recombinant enzyme are evaluated. The potential function of ADAMTS-14 as a physiological aminoprocollagen peptidase in vivo is discussed. [less ▲]

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See detailCloning and characterization of G protein-coupled receptors
Parmentier, M.; Libert, F.; Perret, J. et al

in Advances in Second Messenger and Phosphoprotein Research (1993), 28

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See detailCloning And Characterization Of Mn, A Human Tumor-Associated Protein With A Domain Homologous To Carbonic-Anhydrase And A Putative Helix-Loop-Helix Dna-Binding Segment
Pastorek, J.; Pastorekova, S.; Callebaut, I. et al

in Oncogene (1994), 9(10),

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See detailCloning and distribution of steroid receptor coactivator SRC-1 in quail.
Charlier, Thierry ULg; Lakaye, Bernard ULg; Ball, Gregory F. et al

Poster (2001)

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See detailCloning and embryonic expression of zebrafish PLAG genes.
Pendeville, Helene; Peers, Bernard ULg; Kas, Koen et al

in Gene Expression Patterns (2006), 6(3), 267-76

PLAG transcription factors play important roles in oncogenesis. To date three members of this subfamily of zinc finger proteins have been identified in humans and mice: PLAG1, PLAGL1 and PLAGL2. In this ... [more ▼]

PLAG transcription factors play important roles in oncogenesis. To date three members of this subfamily of zinc finger proteins have been identified in humans and mice: PLAG1, PLAGL1 and PLAGL2. In this study, we identified zebrafish orthologs of PLAG1 and PLAGL2 and a novel member of this family, PLAGX. We examined the temporal expression of these three genes by quantitative real time RT-PCR and found that all three genes are maternally provided, expressed at low level during early somitogenesis and, during late somitogenesis and beyond, PLAG expression increases to reach a plateau level around 5 dpf. Whole mount in situ experiments revealed that PLAG1, PLAGL2 and PLAGX display a similar pattern of expression characterized by a low ubiquitous expression overcame by high expression in some restricted compartments such as the ventricular zone of the brain, the pectoral fin buds, the developing pharyngeal arches and the axial vasculature. We show that this pattern resembles the one observed for the proliferative marker PCNA, suggesting that the PLAG genes are expressed more strongly in zones of active proliferation. This hypothesis was proven for the ventricular zone shown to be a highly proliferative zone using the anti-phosphohistone H3 antibody that detects cells in mitosis. [less ▲]

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See detailCloning and expression analysis of an inducible HSP70 gene from tilapia fish
Molina, Alfredo; Biemar, Frédéric; Muller, Ferenc et al

in FEBS Letters (2000), 474(1), 5-10

We isolated and characterized the tilapia (Oreochromis mossambicus) HSP70 gene, highly homologous to other HSP70 genes. A dramatic increase of tilapia HSP70 mRNA levels was observed after heat shock of ... [more ▼]

We isolated and characterized the tilapia (Oreochromis mossambicus) HSP70 gene, highly homologous to other HSP70 genes. A dramatic increase of tilapia HSP70 mRNA levels was observed after heat shock of whole animals in all organs tested. Reporter constructs were tested for transient expression in carp cells and in microinjected zebrafish embryos. The entire isolated regulatory region (-851/+157) was able to mediate heat shock inducible expression of the reporter gene, with no preference for a particular tissue. Our studies represent the first transcriptional analysis of a HSP70 promoter from fish, revealing a powerful tool to direct controlled, tissue-independent gene expression in fish. [less ▲]

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See detailCloning and Expression Analysis of Cdnas Corresponding to Genes Activated in Cucumber Showing Systemic Acquired Resistance after Bth Treatment
Bovie, C.; Ongena, MARC ULg; Thonart, Philippe ULg et al

in BMC Plant Biology (2004), 4

BACKGROUND: Infection of plants by necrotizing pathogens can lead to the rapid and localized induction of a complex set of defense responses resulting in a restriction of pathogen growth and spread ... [more ▼]

BACKGROUND: Infection of plants by necrotizing pathogens can lead to the rapid and localized induction of a complex set of defense responses resulting in a restriction of pathogen growth and spread. Subsequently, an increase of plant resistance against a broad spectrum of pathogens is observed systemically. This plant immunity is known as Systemic Acquired Resistance. To identify components of the transduction pathway, we cloned and analysed the expression pattern of several mRNAs accumulating in cucumber plants after induction of Systemic Acquired Resistance. RESULTS: We tested on cucumber different compounds known to induce systemic acquired resistance. Among these, BTH (benzo(1,2,3)thiadiazole-7-carbothioic acid S-methyl ester) proved to be very effective. mRNA RT-PCR differential display was used to identify mRNA sequences induced 24 hours after the application of 10 microM BTH to cucumber plants. A cDNA library constructed from cucumber plants sprayed with 10 microM BTH was screened to get corresponding full length cDNAs. Among the identified cDNAs were those coding for a putative ras-related GTP-binding protein, a putative beta-1,4-N-Acetylglucosaminyltranferase III and a putative pathogenesis related protein. The time course of accumulation of the three corresponding mRNAs was analysed by northern blotting in plants treated by BTH or in plants infected by Colletotrichum lagenarium. CONCLUSIONS: The mRNA RT-PCR differential display technique allowed the identification of three genes possibly involved in Systemic Acquired Resistance in cucumber. Pathogenesis-related proteins are known to be involved in plant defence against pathogens. GTP-binding protein and N-acetylglucosaminyltranferase III have been reported to be components of signal transduction pathways in mammals and plants. [less ▲]

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