NICOLAS ROUHIER
285
Documents
Presentation
I am a biochemist interested in basic plant science processes with a particular research focus on structure-function analysis of oxidoreductases and on redox post-translational modifications of proteins.
After a PhD in plant physiology, molecular biology and biochemistry, I have been recruited in 2004 as an assistant professor at the Université of Nancy, obtained an habilitation (HDR) in 2007 before gettting a professor position in 2011 at the newly created Université of Lorraine. For our research projects, we employ biochemical, biophysical, spectroscopic and structural biology approaches with *Populus trichocarpa* and *Arabidopsis thaliana* as model organisms to explore (i) the roles of the antioxidant/detoxification systems in the physiology of plants subject to environmental constraints and (ii) the maturation and roles of iron-sulfur proteins in plant organelles.
- **Redox post-translational modifications of proteins**
In addition to performing structure-function analyses of thioredoxin (TRX) and glutaredoxin (GRX) family members which control most of the reversible oxidative modifications of protein cysteinyl residues, we are characterizing proteins of unknown function possessing one or several conserved CXXC motifs known to be particularly suited for disulfide bond formation but also other redox modifications. We hope to unravel new redox-regulated cellular processes or signaling pathways in poplar and more generally in plants which are controlled by redox reactions such as thiol-disulfide exchanges and understand how the functions of these proteins are controlled at the cellular level by TRX and GRX.
The same questions and strategies apply to sulfurtransferases, enzymes that catalyze the transfer of a sulfur atom from sulfur donors to nucleophilic sulfur acceptors (trans-persulfidation) by forming themselves persulfides. In this context, the intervention of the TRX and glutathione (GSH)/GRX reducing systems in the sulfur signaling/trafficking pathways is examined as both the TRX and GSH/GRX systems might modulate STR function and in particular be central to the formation of hydrogen sulfide, a molecule susceptible to trigger redox signalling cascades in several physiological situations.
**Collaborators: C. Didierjean (Univ. Lorraine), JM. Herrmann (Kaiserslautern), S. Lemaire (CNRS – Sorbonne Université), A. Meyer (Bonn), B. Morgan (Saarbrücken), P. Rey (CEA Cadarache)**
- **Molecular analysis of late steps of Fe-S cluster maturation in organelles**
The general objective here is to understand the functioning of the Fe-S cluster assembly machineries in organelles and more precisely the molecular mechanisms controlling the second step, *i.e.* the trafficking of preformed Fe-S clusters from scaffold proteins (made in the first step) to final acceptors. After focusing for some time on the capacity of several plant GRXs to assemble \[2Fe-2S\] centers into homodimers or into heterodimers with BOLA proteins, we extended our research interest to the functional characterization of the associated protein families (NFUs, SUFA/ISCAs, IBA57s, HCF101/INDH) assumed to participate in the transfer of these Fe-S clusters together with GRX and BOLA. This functional analysis will combine plant genetics and physiology, molecular and structural biology and biochemistry approaches to determine whether these proteins assemble different types of Fe-S clusters and have specific interaction partners and what are the physiological and metabolic consequences of deleting these genes for plant development and physiology. Particular emphasis is given to the maturation of the photosynthetic and respiratory electron transfer complexes.
**Collaborators: J. Balk (JIC, Norwich), MK. Johnson (Athens, USA), O. Keech (Umea), A. Meyer (Bonn), U. Mühlenhoff (Marburg), C. Remacle (Liège), F. Vignols (BPMP Montpellier)**
- **Structure-Function analysis of glutathione transferases**
Glutathione transferases (GSTs) constitute large multigenic families of more than 50 genes in terrestrial plants. They have three major biochemical functions. Those having a serinyl residue in their catalytic site (Ser-GSTs) possess GSH-conjugation activity. This catalytic activity is often described as required for xenobiotic detoxification. Those having a cysteinyl residue in their catalytic site (Cys-GSTs) possess the ability to catalyze the reverse reaction, *i.e.*, the removal of glutathione from a number of structurally different heterocyclic molecules. The third function is the binding and/or transport of molecules/specialized metabolites without catalytic transformation, so-called ligandin function. It can be performed both by Ser-GSTs and Cys-GSTs. Although they are present in all kingdoms, the physiological functions of most plant GSTs remain elusive.
The current work is centered on the structure-function analysis of specific classes of glutathione transferases, focusing in particular on the isolation and identification of physiological substrates explaining their implication in biotic or abiotic stress responses. In particular, we are interested in GSTs regulated during poplar leaf infection by the rust fungus *Melampsora larici-populina* or regulated during iron starvation in *A. thaliana*.
The strategies used are to perform pull-down and affinity chromatographies, inhibition kinetics and thermal denaturation combining fluorescent probes and model ligands or plant extracts that are eventually separated by liquid ichromatography. We will not only focus on the previously characterized Cys-GSTs (lambda GSTs and glutathionyl hydroquinone reductases) but also on less explored or newly identified GST families. The nature (ligandin *vs* catalytic transformation) and the strength of the interactions will be validated by measuring binding constants using glutathionylated and non-glutathionylated molecules using biophycal approaches (ITC, Switchsense, fluorimetry). In parallel, the structure of GST alone or in complex with their ligand/substrate will be tentatively solved. Overall, we expect to decipher new intermediary steps of xenobiotic detoxification or of metabolic pathways requiring glutathione-conjugating or transport activities. New research questions have emerged recently from the observation that tau GST are able to bind porphyrins and hemes b in particular.
**Collaborators: C. Didierjean (Univ. Lorraine)**, **C. Dubos (BPMP, Montpellier), S. Lemaire (CNRS – Sorbonne Université)**
**Grants**
-2019-2022 : **Projet ANR-2018**: MITOGLU: Dissecting the molecular interactions of mitochondrial glutaredoxin S15 in plants (coordinators N. Rouhier/A. Meyer)
-2018-2021 : **Projet Lorraine Université Excellence**, PEPS Mirabelle + : FeSprot: Maturation and structure-function analyses of iron-sulfur proteins (coordinator N. Rouhier)
– 2018-2021: **Projet ANR 2017**: MOBIFER: Dynamics of coumarin secretion by plant roots into the soil in the context of iron nutrition (coordinator C. Dubos)
– 2014-2017: **Projet ANR 2013**: Fe-S traffic: The cellular trafficking of iron-sulfur (Fe-S) clusters in plants (coordinator N. Rouhier)
– 2011-2013: **Projet ANR 2010**: FIRES: Functions of glutaredoxins in Iron and REdox sensing and Signaling in plants (coordinator N. Rouhier)
– 2009-2011: **Projet INRA/FORMAS:** The role of organellar glutaredoxins in iron-sulfur cluster biogenesis in plants: an interactomic study for the identification of protein partners (G. Wingsle (Umea Plant Science center, sweden)/ coordinator N. Rouhier)
– 2008-2010: **Projet ANR Jeune Chercheur**: The role of organellar CGFS containing glutaredoxins in iron-sulfur clusters assembly and biogenesis in plants (coordinator N. Rouhier)
– 2006-2008: **Projet ANR, Réseau de Génomique végétale-Genoplante 2010**: GNP05010P: Repair mechanisms of oxidized proteins in chloroplasts of *Arabidopsis thaliana* (coordinator Pascal Rey)
Publications
Monitoring intracellular dynamics of cysteine with the genetically-encoded biosensor CyReBGDR NADPH Oxydase-Stress Oxydant & Club Oxydase, Nov 2023, Orsay, France
Conference papers
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Monitoring intracellular dynamics of cysteine with the genetically-encoded biosensor CyReBCompartmentalized redox biology, meeting of the study group redox biology of the German Society for Biochemistry and Molecular Biology, Sep 2023, Düsseldorf, Germany
Conference papers
hal-04300871v1
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Are Trx-like 3 and 4 new players in the thiol-based redox metabolism in the chloroplast ?Compartmentalized redox biology, Jan Riemer & Carsten Berndt, Sep 2023, Düsseldorf, Germany
Conference papers
hal-04229620v1
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Role of the chloroplastic iron-sulfur transfer protein NFU1 in the microalga Chlamydomonas reinhardtiiBrussels Institute for Advanced Studies - Photosynthesis and primary production, Christian Hermans, Feb 2022, e-conference, Belgium
Conference papers
hal-04176818v1
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3-mercaptopyruvate sulfurtransferases: new actors in sulfur trafficking and H2S mediated-redox signalling in plants?Redox Biology Congress 2022 Oxidative stress, redox biology and antioxidants from plants to humans, Aug 2022, Ghent, Belgium
Conference papers
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Rhodanese domain-containing sulfurtransferase family: a yet mysterious component of sulfur trafficking in plants12th International Plant Sulfur Workshop, Jul 2022, London, Canada
Conference papers
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Maturation of iron-sulfur proteins in chloroplasts: focus on the NFU transfer proteinsSPS “Symposium on redox regulation and carbon-nitrogen interactions in plants. A tribute to Pierre Gadal”, Jun 2022, Gif sur Yvette, France
Conference papers
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Gene atlas of iron-containing proteins (ironOME) in Arabidopsis thalianaIron Nutrition and Interactions in Plants, Jul 2022, Reims, France
Conference papers
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Deciphering the biochemical relationships between cysteine desulfurase and sulfurtransferase protein families in sulfur trafficking12th International Plant Sulfur Workshop, Jul 2022, London, Canada
Conference papers
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The chloroplastic iron-sulfur transfer protein NFU1 is required for chlorophyll synthesis in the dark and fermentation in the green microalga Chlamydomonas reinhardtiiIron Sulfur Proteins: Biogenesis, Regulation and Function meeting, Sep 2022, Sainte Maxime, France
Conference papers
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Redox properties of the plant atypical thioredoxin DCC1.Redox Biology Congress, Aug 2022, Ghent, Belgium
Conference papers
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The critical role of glutaredoxin S15 for the maturation of iron-sulfur proteins in mitochondria.12th International Conference for Plant Mitochondrial Biology 2022, May 2022, Malmö, Sweden
Conference papers
hal-03705989v1
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The Fe-S cluster assembly factors NFU4 and NFU5 are primarily required for protein lipoylation in plant mitochondria.Congrès du Groupe Français de Bioénergétique, Sep 2021, online seminars, France
Conference papers
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Role of the chloroplastic NFU3 iron-sulfur transfer protein in the microalga Chlamydomonas reinhardtii.International Bioenergy and environment congress, Feb 2021, e-conference, France
Conference papers
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Towards the identification of novel metal- or thiol-based regulated proteins in plantsInternational Symposium of the DFG Priority Program 1710 Dynamics of Thiol-based Redox Switches in Cellular Physiology, Mar 2021, e-conference, Germany
Conference papers
hal-03197988v1
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The role of chloroplastic NFU in the green alga Chlamydomonas reinhardtiiOnline seminar series on “Iron-sulfur protein biogenesis 2021”, Jun 2021, e-seminar, France
Conference papers
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Maturation of Fe-S proteins in plant organelles.12th congress of the French Society of Plant Biology, Jul 2019, Strasbourg, France
Conference papers
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Sulfur trafficking and H2S-mediated redox signaling in plants: roles of Arabidopsis 3-MP sulfurtransferasesSPP1710 Conference Thiol-based switches and redox regulation – from microbes to men, Sep 2019, Sant Feliu de Guixols, Spain
Conference papers
hal-03000210v1
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Are the ubiquitous mitochondrial glutaredoxin-like proteins of 12 kDa (MGP12) true glutaredoxins?Thiol-based switches and redox regulation – from microbes to men, Sep 2019, San Feliu de Guixols, Spain
Conference papers
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Elucidating the function of the chloroplastic glutaredoxin S16 of Arabidopsis thaliana.12th Congress of the International Plant Molecular Biology, Aug 2018, Montpellier, France
Conference papers
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The maturation of Fe-S proteins in chloroplasts and mitochondriaJacques Monod conference “Retrograde signalling from endosymbiotic organelles”, Oct 2018, Roscoff, France
Conference papers
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Functional characterization of mitochondrial NFU proteins in plants8th International Conference on Iron-Sulfur Cluster Biogenesis and Regulation, May 2018, Madison, United States
Conference papers
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Dynamics of coumarin secretion by plant roots into the soil to improve iron nutritionXVIII International Symposium on Iron Nutrition and Interactions in Plants, Jul 2018, Taipei, Taiwan
Conference papers
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Deciphering the biochemical properties of bacterial rhodanese fusion proteins.Fourth meeting of the study group redox biology of the German Society for Biochemistry and Molecular Biology, Sep 2018, Berlin, Germany
Conference papers
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Arabidopsis thaliana GRXS16, a multifaceted glutaredoxin in chloroplasts.3rd Redox Biology Symposium, Redox regulation: Historical background and future developments, Mar 2017, Nancy, France
Conference papers
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Who are the partners of mitochondrial Arabidopsis thaliana GRXS15 in trafficking of iron-sulfur clusters?Redox meeting and Conversation, Mar 2017, Vandoeuvre Lès Nancy, France
Conference papers
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How are Fe-S cofactors and proteins assembled in plant cells? focus on NFUs, involved in the late steps of the maturation process in organellesDays of the French society of photosynthesis, Jun 2017, Paris, France
Conference papers
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The chloroplastic glutaredoxin S16: a plant-specific, two-domain protein with multiple biochemical properties.EMBO Conference on Redox Biology, Jul 2017, Moscou, Russia
Conference papers
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Deciphering the late-steps of FeS cluster assembly in plant organelles: focus on NFUs.Kick off meeting of the COST action CA15133 FeSBioNET “The biogenesis of iron-sulfur proteins: from cellular biology to molecular aspects”., Sep 2016, Patras, Greece
Conference papers
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Deciphering the molecular mechanisms of sulfur trafficking in plants.hird meeting of the study group redox biology of the German Society for Biochemistry and Molecular Biology, Jul 2016, Düsseldorf, Germany
Conference papers
hal-03000214v1
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Structure-function analyses of plant glutathione transferases with catalytic cysteine residuesTheoretical and Practical Course “Redox Chemistry and Biology of Thiols” and associated symposium., Mar 2015, montevideo, Uruguay
Conference papers
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The highly expanded thioredoxin and glutaredoxin classes in plants: specificities in the regeneration systems of enzymes using a sulfenic acid chemistryTheoretical and Practical Course “Redox Chemistry and Biology of Thiols” and associated symposium, Mar 2015, Montevideo, Uruguay
Conference papers
hal-03198450v1
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Structure-function analysis of cysteine-containing glutathione transferases (GSTs) from Populus trichocarpaThiol-based redox switches in life sciences, EMBO-ESF meeting, Sep 2015, san feliu de guixols, Spain
Conference papers
hal-03198459v1
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Exploring the partners of late-acting Fe-S transfer proteins belonging to the mitochondrial ISC machinery in Arabidopsis thaliana.Labex Mitocross 2015: mitochondria at the crossroad, Sep 2015, Strasbourg, France
Conference papers
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The multiple facets of glutaredoxins: one fold, several partners and functions12th International Conference on Reactive Oxygen and Nitrogen Species in Plants: from model systems to field, Jun 2015, verone, Italy
Conference papers
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Plastidial Nfu2 and Nfu3 are crucial for a proper root and shoot development in Arabidopsis thaliana7th International Conference on Iron-Sulfur Cluster Biogenesis and Regulation., Jun 2015, bergame, Italy
Conference papers
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Biochemical, spectroscopic and structural insights into BolA-glutaredoxin complexesIUBMB Symposium FeS 2015 - Iron Sulfur Cluster Biogenesis and Regulation, Jun 2015, Bergamo, Italy
Conference papers
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Biochemical, spectroscopic and structural insights into BolA-glutaredoxin complexes.Gordon research conference “Thiol-based redox regulation and signalling”., Jul 2014, girone, Spain
Conference papers
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Biochemical, spectroscopic and structural insights into BolA-glutaredoxin complexes.IJB Symposium «Molecular Sciences, Facing Up to Major Societal Challenges»., Oct 2014, Nancy, France
Conference papers
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Genomics, transcriptomics and effectoromics of the poplar leaf rust fungus Melampsora larici-populinaIUFRO Joint Meeting on Genetics of Five-Needle Pines and Rusts in Mountain Landscapes, Jun 2014, Fort Collins, United States
Conference papers
hal-02801804v1
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Plastidial glutaredoxins: glutathione-dependent enzymes involved in detoxification, redox signalling and iron-sulfur cluster biogenesis.9th International Workshop - Sulfur Metabolism in Plants., Apr 2014, freiburg, Germany
Conference papers
hal-03198438v1
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Monothiol glutaredoxin and BolA in plants: two proteins with multiple interaction modes.9th International Workshop on Sulfur Metabolism in Plants: Molecular Physiology and Ecophysiology of Sulfur, Apr 2014, Freiburg, Germany
Conference papers
hal-03000377v1
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Biochemical, spectroscopic and structural insights into glutaredoxins and their multiple types of interactions with BolAs2nd Symposium of the DFG Priority Program SPP 1710 “Dynamics of Thiol-based Redox Switches in Cellular Physiology”, Sep 2014, Heidelberg, Germany
Conference papers
hal-03198444v1
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Investigation of the role of glutaredoxins and NFU proteins for the plastidial iron-sulfur cluster assembly machinary5th International Meeting of the Institute of Metals in Biology of Grenoble (IMBG)/Metal Homeostasis, Sep 2012, Autrans, France
Conference papers
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Les apports de la génomique pour l’étude des familles multigéniques : exemple des thaumatin-like proteins (TLPs) chez le peuplierSéminaire de l’Ecole Doctorale RP2E. Biodiversité, Ingénierie écologique, Jan 2011, Vandoeuvre-lès-Nancy, France. 8 p
Conference papers
hal-02746248v1
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Biochemical characterization of thioredoxin-related protein Clot/TRP14 of Populus trichocarpaCompartmentalized redox biology, meeting of the study group redox biology of the German Society for Biochemistry and Molecular Biology, Sep 2023, Düsseldorf, Germany
Conference poster
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Development of a genetically-encoded biosensor to monitor an H2S-related metaboliteSociety for Redox Biology and Medicine (SfRBM) 30th Annual Conference and Society for Free Radical Research International (SFRRI) 21st Biennial Congress, Nov 2023, Punta del Este, Uruguay, Uruguay
Conference poster
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The complex redox regulation of the SAL1 phosphatase present in plant organellesThiol oxidation in biology: Biochemical mechanisms to physiological outcomes, Oct 2022, sant feliu de guixols, Spain
Conference poster
hal-03820252v1
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Iron-sulfur cluster formation and transfer using plant mitochondrial glutaredoxin-BOLA complexes.Iron Sulfur Proteins: Biogenesis, Regulation and Function meeting., Sep 2022, Sainte-maxime, France
Conference poster
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Exploiting a bacterial cysteine desulfurase-sulfurtransferase fusion to develop a roGFP2-based biosensor for free cysteineRedox Biology Congress 2022 Oxidative stress, redox biology and antioxidants from plants to humans, Aug 2022, Ghent, Belgium
Conference poster
hal-03775303v1
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Maturation of iron-sulfur-containing subunits of respiratory complex I in plantsIron Sulfur Proteins: Biogenesis, Regulation and Function meeting, Sep 2022, Sainte Maxime, France
Conference poster
hal-03781848v1
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Sulfurtransferase-mediated formation of H2S and persulfidation in Arabidopsis thalianaBotanik-Tagung International Conference of the German Society for Plant Sciences, Aug 2022, Bonn, Germany
Conference poster
hal-03775320v1
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Unravelling the role of an atypical mitochondrial ferredoxin from Arabidopsis thaliana.Congrès du Groupe Français de Bioénergétique, Sep 2021, online seminar, France
Conference poster
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Shedding light on a H2S generating pathway in ArabidopsisS-Bio 2021 Glucosinolate Joint meeting for plant and human sulfur biology and glucosinolates, Sep 2021, Seville, Spain
Conference poster
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Identification of client iron-sulfur proteins of the chloroplastic NFU3 transfer protein from Chlamydomonas reinhardtii.International Bioenergy and environment congress, Feb 2021, online seminar, France
Conference poster
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Investigation of cellular interactions and biochemical characterization of the chloroplastic NFU1 transfer protein from Chlamydomonas reinhardtii.Chlamy 2020+1. 19th International Conference on the Cell and Molecular Biology of Chlamydomonas., Aug 2021, Six-Fours-les-Plages, France
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hal-03707828v1
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Deciphering the biochemical relationships between cysteine desulfurase and sulfurtransferase protein families in sulfur traffickingS-Bio 2021 Glucosinolate Joint meeting for plant and human sulfur biology and glucosinolates, Sep 2021, Seville, Spain
Conference poster
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Dynamics of coumarin secretion by plant roots into the soil to improve iron nutrition12ème Congrès de la Société Française de Biologie Végétale SFBV, Jul 2019, Strasbourg, France
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