References of "Remacle, Claire"
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See detailFunctional distribution and dynamics of Arabidopsis SR splicing factors in living plant cells
Tillemans, Vinciane ULg; Dispa, Laurence ULg; Remacle, Claire ULg et al

in Plant Journal (The) (2005), 41(4), 567-582

Serine/arginine-rich (SR) proteins constitute an important class of splicing regulators in higher eukaryotes that share a modular structure consisting of one or two N-terminal RNA recognition motif (RRM ... [more ▼]

Serine/arginine-rich (SR) proteins constitute an important class of splicing regulators in higher eukaryotes that share a modular structure consisting of one or two N-terminal RNA recognition motif (RRM) domains and a C-terminal RS-rich domain. Herein, we have investigated the in vivo functional distribution of Arabidopsis SR factors. Agrobacterium-mediated transient transformation revealed nuclear speckled distribution and the overall colocalization of fluorescent protein (FP)-tagged SR factors in both tobacco and Arabidopsis cells. Their overall colocalization in larger nucleoplasmic domains was further observed after transcriptional and phosphorylation/dephosphorylation inhibition, indicating a close functional association between SR factors, independent of their phosphorylation state. Furthermore, we demonstrated in vivo the conserved role of the RS and RRM domains in the efficient targeting of Arabidopsis SR proteins to nuclear speckles by using a series of structural domain-deleted mutants of atRSp31 and atRSZp22. We suggest additional roles of RS domain such as the shuttling of atRSZp22 between nucleoplasm and nucleolus through its phosphorylation level. The coexpression of deletion mutants with wild-type SR proteins revealed potential complex associations between them. Fluorescence recovery after photobleaching demonstrated similar dynamic properties of SR factors in both tobacco transiently expressing cells and Arabidopsis transgenics. Cell cycle phase-dependent organization of FP-tagged SR proteins was observed in living tobacco BY-2 cells. We showed that atRSp31 is degraded at metaphase by fluorescence quantification. SR proteins also localized within small foci at anaphase. These results demonstrate interesting related features as well as potentially important differences between plant and animal SR proteins. [less ▲]

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See detailThe mitochondrial oxidative phosphorylation proteome of Chlamydomonas reinhardtii deduced from the genome sequencing project
Cardol, Pierre ULg; Gonzalez-Halphen, Diego; Reyes-Prieto, Adrian et al

in Plant Physiology (2005), 137(2), 447-459

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See detailStructural organization of mitochondrial human complex I: role of the ND4 and ND5 mitochondria-encoded subunits and interaction with prohibitin
Bourges, I.; Ramus, C.; de Camaret, B. M. et al

in Biochemical Journal (2004), 383(Part 3), 491-499

Mitochondria-encoded ND (NADH dehydrogenase) subunits, as components of the hydrophobic part of complex I, are essential for NADH:ubiquinone oxidoreductase activity. Mutations or lack of expression of ... [more ▼]

Mitochondria-encoded ND (NADH dehydrogenase) subunits, as components of the hydrophobic part of complex I, are essential for NADH:ubiquinone oxidoreductase activity. Mutations or lack of expression of these subunits have significant pathogenic consequences in humans. However, the way these events affect complex I assembly is poorly documented. To understand the effects of particular mutations in ND subunits on complex I assembly, we studied four human cell lines: ND4 non-expressing cells, ND5 non-expressing cells, and rhodegrees cells that do not express any ND subunits, in comparison with normal complex I control cells. In control cells. all the seven analysed nuclear-encoded complex I subunits Were found to be attached to the mitochondrial inner membrane, except for the 24 kDa subunit, which was nearly equally partitioned between the membranes and the matrix. Absence of a single ND subunit, or even all the seven ND subunits, caused no major changes in the nuclear-encoded complex I subunit content of mitochondria. However, in cells lacking ND4 or ND5, very low amounts of 24 kDa subunit were found associated with the membranes, whereas most of the other nuclear-encoded subunits remained attached. In contrast, membrane association of most of the nuclear subunits was significantly reduced in the absence of all seven ND proteins. Immunopurification detected several subcomplexes. One of these, containing the 23, 30 and 49 kDa subunits, also contained prohibitin. This is the first description of prohibitin interaction with complex I subunits and suggests that this protein might play a role in the assembly or degradation of mitochondrial complex I. [less ▲]

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See detailHigher plant-like subunit composition of mitochondrial complex I from Chlamydomonas reinhardtii: 31 conserved components among eukaryotes
Cardol, Pierre ULg; Vanrobaeys, F.; Devreese, B. et al

in Biochimica et Biophysica Acta-Bioenergetics (2004), 1658(3), 212-224

The rotenone-sensitive NADH:ubiquinone oxidoreductase (complex I) is the most intricate membrane-bound enzyme of the mitochondrial respiratory chain. Notably the bovine enzyme comprises up to 46 subunits ... [more ▼]

The rotenone-sensitive NADH:ubiquinone oxidoreductase (complex I) is the most intricate membrane-bound enzyme of the mitochondrial respiratory chain. Notably the bovine enzyme comprises up to 46 subunits, while 27 subunits could be considered as widely conserved among eukaryotic complex I. By combining proteomic and genomic approaches, we characterized the complex I composition from the unicellular green alga Chlamydomonas reinhardtii. After purification by blue-native polyacrylamide gel electrophoresis (BN-PAGE), constitutive subunits were analyzed by SDS-PAGE coupled to tandem mass spectrometry (MS) that allowed the identification of 30 proteins. We compared the known complex I components from higher plants, mammals, nematodes and fungi with this MS data set and the translated sequences from the algal genome project. This revealed that the Chlamydomonas complex I is likely composed of 42 proteins, for a total molecular mass of about 970 kDa. In addition to the 27 typical components, we have identified four new complex I subunit families (bovine ESSS, PFFD, B16.6, B12 homologues), extending the number of widely conserved eukaryote complex I components to 31. In parallel, our analysis showed that a variable number of subunits appears to be specific to each eukaryotic kingdom (animals, fungi or plants). Protein sequence divergence in these kingdom-specific sets is significant and currently we cannot exclude the possibility that homology between them exists, but has not yet been detected. (C) 2004 Elsevier B.V. All rights reserved. [less ▲]

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See detailImpact of a mutation in the mitochondrial LSU rRNA gene from Chlamydomonas reinhardtii on the activity and the assembly of respiratory-chain complexes
Remacle, Claire ULg; Gloire, Geoffrey ULg; Cardol, Pierre ULg et al

in Current Genetics (2004), 45(5), 323-330

Two substitutions A1090G and A1098C (together called the m mutation) located in the conserved GTPase domain of the mitochondrial LSU rRNA gene were recently shown to weakly compensate for the phenotypical ... [more ▼]

Two substitutions A1090G and A1098C (together called the m mutation) located in the conserved GTPase domain of the mitochondrial LSU rRNA gene were recently shown to weakly compensate for the phenotypical effect of a -1T frameshift mutation in the mitochondrial cox1 gene of C. reinhardtii. In order to analyze the impact of the m mutation on the mitochondrial translational machinery, a strain carrying the m mutation but wild-type for the cox1 gene was isolated. We found that the growth and the respiratory rate of the m mutant were affected and that the activities of complexes I, III, and IV, all containing mitochondria-encoded subunits, were lowered. In contrast the activities of complex II and of the alternative oxidase, both encoded exclusively by the nuclear genome, were not modified. The steady-state levels of complex I enzyme and of several components of the respiratory complexes I, III, and IV were also reduced in the mutant. We moreover showed that m did not suppress other frameshift or UGA stop mutations which affect mitochondrial genes. [less ▲]

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See detailProteomic and genomic analysis of Chlamydomonas reinhardtii complex 1: conserved components in eukaryotes
Cardol, Pierre ULg; Vanrobaeys, F.; Devreese, B. et al

in Biochimica et Biophysica Acta-Bioenergetics (2004), 1657(Suppl. 1), 41-42

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See detailPhotosynthesis and state transitions in mitochondrial mutants of Chlamydomonas reinhardtii affected in respiration
Cardol, Pierre ULg; Gloire, Geoffrey ULg; Havaux, M. et al

in Plant Physiology (2003), 133(4), 2010-2020

Photosynthetic activities were analyzed in Chlamydomonas reinhardtii mitochondrial mutants affected in different complexes (I, III, IV, I + III, and I + IV) of the respiratory chain. Oxygen evolution ... [more ▼]

Photosynthetic activities were analyzed in Chlamydomonas reinhardtii mitochondrial mutants affected in different complexes (I, III, IV, I + III, and I + IV) of the respiratory chain. Oxygen evolution curves showed a positive relationship between the apparent yield of photosynthetic linear electron transport and the number of active proton-pumping sites in mitochondria. Although no significant alterations of the quantitative relationships between major photosynthetic complexes were found in the mutants, 77 K fluorescence spectra showed a preferential excitation of photosystem I (PSI) compared with wild type, which was indicative of a shift toward state 2. This effect was correlated with high levels of phosphorylation of light-harvesting complex II polypeptides, indicating the preferential association of light-harvesting complex II with PSI. The transition to state 1 occurred in untreated wild-type cells exposed to PSI light or in 3-(3,4-dichlorophenyl)-1,1-dimethylurea-treated cells exposed to white light. In mutants of the cytochrome pathway and in double mutants, this transition was only observed in white light in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea. This suggests higher rates of non-photochemical plastoquinone reduction through the chlororespiratory pathway, which was confirmed by measurements of the complementary area above the fluorescence induction curve in dark-adapted cells. Photo-acoustic measurements of energy storage by PSI showed a stimulation of PSI-driven cyclic electron flow in the most affected mutants. The present results demonstrate that in C. reinhardtii mutants, permanent defects in the mitochondrial electron transport chain stabilize state 2, which favors cyclic over linear electron transport in the chloroplast. [less ▲]

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See detailImpact of mutations affecting ND mitochondria-encoded Subunits on the activity and assembly of complex I in chlamydomonas. Implication for the structural organization of the enzyme
Cardol, Pierre ULg; Matagne, René-Fernand ULg; Remacle, Claire ULg

in Journal of Molecular Biology (2002), 319(5), 1211-1221

The mitochondrial rotenone-sensitive NADH:ubiquinone oxidoreductase (complex 1) comprises more than 35 subunits, the majority of which are encoded by the nucleus. In Chlamydomonas reinhardtii, only five ... [more ▼]

The mitochondrial rotenone-sensitive NADH:ubiquinone oxidoreductase (complex 1) comprises more than 35 subunits, the majority of which are encoded by the nucleus. In Chlamydomonas reinhardtii, only five components (ND1, ND2, ND4, ND5 and ND6) are coded for by the mitochondrial genome. Here, we characterize two mitochondrial mutants (dum5 and dum17) showing strong reduction or inactivation of complex I activity: dum5 is a IT deletion in the 3' UTR of nd5 whereas dum17 is a IT deletion in the coding sequence of nd6. The impact of these mutations and of mutations affecting nd1, nd4 and nd4/nd5 genes on the assembly of complex I is investigated. After separation of the respiratory complexes by blue native (BN)-PAGE or sucrose gradient centrifugation, we demonstrate that the absence of intact ND1 or ND6 subunit prevents the assembly of the 850 kDa whole complex, whereas the loss of ND4 or ND4/ND5 leads to the formation of a subcomplex of 650 kDa present in reduced amount. The implications of our findings for the possible role of these ND subunits on the activity of complex I and for the structural organization of the membrane arm of the enzyme are discussed. In mitochondria from all the strains analyzed, we moreover detected a 160210 kDa fragment comprising the hydrophilic 49 kDa and 76 kDa subunits of the complex I peripheral arm and showing NADH dehydrogenase activity. (C) 2002 Elsevier Science Ltd. All rights reserved. [less ▲]

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See detailThe genetics and molecular biology of mitochondria in Chlamydomonas
Matagne, René-Fernand ULg; Remacle, Claire ULg

in Pandalai, S. (Ed.) Recent Research Development in Plant Biology (2002)

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See detailStructure of the Telomeric Ends of Mt DNA, Transcriptional Analysis and Complex I Assembly in the Dum24 Mitochondrial Mutant of Chlamydomonas Reinhardtii
Duby, Franceline ULg; Cardol, Pierre ULg; Matagne, René-Fernand ULg et al

in Molecular Genetics & Genomics (2001), 266(1), 109-14

The dum24 mutant of Chlamydomonas reinhardtii contains four types of altered mitochondrial linear genomes: two types of deleted monomers and two types of dimers resulting from fusions between some ... [more ▼]

The dum24 mutant of Chlamydomonas reinhardtii contains four types of altered mitochondrial linear genomes: two types of deleted monomers and two types of dimers resulting from fusions between some monomers via their deleted ends. All molecules lack at least cob, nd4 and the 3' end of nd5, three adjacent genes located in the left part of the genome. We present evidence showing that in dum24, as in other deletion mutants, the deletions extend to the left telomeric end, and propose that the only replicative forms in the mutants are the dimeric DNA molecules that possess intact telomeric structures at both ends. Two abnormally large transcripts produced from chimeric genes are detected in dum24, which throws some light on the location of potential promoter sequences and processing signals in the mitochondrial genome. Using BN-PAGE analysis and immunological methods to detect complex I, we further show that dum24 mitochondria do not possess the normal multimeric complex I (850 kDa), but produce a smaller, partially assembled, complex (650 kDa), demonstrating a role for ND4 and/or ND5 subunits(s) in complex I assembly. [less ▲]

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See detailMutations Inactivating Mitochondrial Genes in Chlamydomonas Reinhardtii
Remacle, Claire ULg; Duby, Franceline ULg; Cardol, Pierre ULg et al

in Biochemical Society Transactions (2001), 29(Pt 4), 442-6

Chlamydomonas reinhardtii is now becoming a useful model for the study of mitochondrial genetics in a photosynthetic organism. The small (15.8 kb) mitochondrial genome C. reinhardtii has been sequenced ... [more ▼]

Chlamydomonas reinhardtii is now becoming a useful model for the study of mitochondrial genetics in a photosynthetic organism. The small (15.8 kb) mitochondrial genome C. reinhardtii has been sequenced completely and all the genes have been identified. Several mutants inactivated in mitochondrial genes encoding components of the respiratory complexes I, III and IV have been characterized at the molecular level. Assembly of complex I in several mutant strains and mapping of mitochondrial mutations by recombinational analysis are also described. [less ▲]

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See detailMutants of Chlamydomonas reinhardtii deficient in mitochondrial complex I: Characterization of two mutations affecting the nd1 coding sequence
Remacle, Claire ULg; Baurain, Denis ULg; Cardol, Pierre ULg et al

in Genetics (2001), 158(3), 1051-60

The mitochondrial rotenone-sensitive NADH:ubiquinone oxidoreductase (complex I) comprises more than 30 subunits, the majority of which are encoded by the nucleus. In Chlamydomonas reinhardtii, only five ... [more ▼]

The mitochondrial rotenone-sensitive NADH:ubiquinone oxidoreductase (complex I) comprises more than 30 subunits, the majority of which are encoded by the nucleus. In Chlamydomonas reinhardtii, only five components of complex I are coded for by mitochondrial genes. Three mutants deprived of complex I activity and displaying slow growth in the dark were isolated after mutagenic treatment with acriflavine. A genetical analysis demonstrated that two mutations (dum20 and dum25) affect the mitochondrial genome whereas the third mutation (dn26) is of nuclear origin. Recombinational analyses showed that dum20 and dum25 are closely linked on the genetic map of the mitochondrial genome and could affect the nd1 gene. A sequencing analysis confirmed this conclusion: dum20 is a deletion of one T at codon 243 of nd1; dum25 corresponds to a 6-bp deletion that eliminates two amino acids located in a very conserved hydrophilic segment of the protein. [less ▲]

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