Norbert Rolland

Norbert Rolland
French National Centre for Scientific Research | CNRS · Institute of Biological Sciences

PhD

About

121
Publications
29,855
Reads
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7,077
Citations
Citations since 2016
28 Research Items
2710 Citations
20162017201820192020202120220100200300400500
20162017201820192020202120220100200300400500
20162017201820192020202120220100200300400500
20162017201820192020202120220100200300400500
Introduction
PhD in 1992 (Univ. Grenoble, France), lab of Prof. Roland Douce (Lyon, France): metabolism, biochemistry, enzymology - Post-doc (EMBO) in 1993-1994, lab of Prof. Jean-David Rochaix (Univ. Geneva, Switzerland): photosynthesis, molecular biology, genetics - Junior CNRS Scientist position in 1995 (Univ. Grenoble, Cell and Plant Physiology lab, headed by Jacques Joyard). Team leader since 1999 and CNRS Research Director since 2004 - Deputy Head (2003-2010) then Head (2010-2017) of the Cell and Plant Physiology lab in Grenoble - Deputy Head of the Plant Biology and Breeding Division at INRA (since 2017). Research focused on chloroplast functions, metabolism, biochemistry, biogenesis, transporters, proteomics... in both higher plants and unicellular algae.
Additional affiliations
April 2017 - present
French National Institute for Agriculture, Food, and Environment (INRAE)
Position
  • Head of Department
June 2005 - June 2009
French National Centre for Scientific Research
Position
  • Head (2005-2009) of the CNRS GDR (National Research Network) on Photosynthesis
January 2004 - present
French National Centre for Scientific Research
Position
  • Managing Director
Education
January 1993 - December 1994
University of Geneva
Field of study
  • Molecular Biology and Genetics
October 1989 - December 1992
University Joseph Fourier - Grenoble 1
Field of study
  • Cellular and Molecular Biology

Publications

Publications (121)
Article
Full-text available
The development of chloroplasts and the integration of their function within a plant cell rely on the presence of a complex biochemical machinery located within their limiting envelope membranes. To provide the most exhaustive view of the protein repertoire of chloroplast envelope membranes, we analyzed this membrane system using proteomics. To thi...
Article
Full-text available
Recent advances in the proteomics field have allowed a series of high throughput experiments to be conducted on chloroplast samples, and the data are available in several public databases. However, the accurate localization of many chloroplast proteins often remains hypothetical. This is especially true for envelope proteins. We went a step further...
Article
Full-text available
Plastids are semiautonomous organelles derived from cyanobacterial ancestors. Following endosymbiosis, plastids have evolved to optimize their functions, thereby limiting metabolic redundancy with other cell compartments. Contemporary plastids have also recruited proteins produced by the nuclear genome of the host cell. In addition, many genes acqu...
Article
Full-text available
The chloroplast is a major plant cell organelle that fulfills essential metabolic and biosynthetic functions. Located at the interface between the chloroplast and other cell compartments, the chloroplast envelope system is a strategic barrier controlling the exchange of ions, metabolites and proteins, thus regulating essential metabolic functions (...
Article
Full-text available
Cell compartmentalization is an essential process by which eukaryotic cells separate and control biological processes. While calmodulins are well known to regulate catalytic properties of their targets, we show here their involvement in the subcellular location of two plant proteins. Both proteins exhibit a dual location, namely in the cytosol in a...
Article
Full-text available
The world needs more than double its current agricultural productivity by 2050 to produce enough food and feed, as well as to provide feedstock for the bioeconomy. These future increases will not only need to be sustainable but also need to compromise the nutritional quality, and ideally also need to decrease greenhouse gas emissions and increase c...
Article
Full-text available
Global warming and drought stress are expected to have a negative impact on agricultural productivity. Desiccation-tolerant species, which are able to tolerate the almost complete desiccation of their vegetative tissues, are appropriate models to study extreme drought tolerance and identify novel approaches to improve the resistance of crops to dro...
Article
Full-text available
The realization of the full objectives of international policies targeting global food security and climate change mitigation, including the United Nation’s Sustainable Development Goals, the Paris Climate Agreement COP21 and the European Green Deal, requires that we (i) sustainably increase the yield, nutritional quality and biodiversity of major...
Article
Full-text available
The multinational Arabidopsis research community is highly collaborative and over the past thirty years these activities have been documented by the Multinational Arabidopsis Steering Committee (MASC). Here, we (a) highlight recent research advances made with the reference plant Arabidopsis thaliana ; (b) provide summaries from recent reports submi...
Article
Full-text available
Chloroplasts are major components of plant cells. Such plastids fulfill many crucial functions, such as assimilation of carbon, sulfur and nitrogen as well as synthesis of essential metabolites. These organelles consist of the following three key sub-compartments. The envelope, characterized by two membranes, surrounds the organelle and controls th...
Chapter
Plastids contain several key subcompartments. The two limiting envelope membranes (inner and outer membrane of the plastid envelope with an intermembrane space between), an aqueous phase (stroma), and an internal membrane system terms (thylakoids) formed of flat compressed vesicles (grana) and more light structures (lamellae). The thylakoid vesicle...
Chapter
Plastids are semiautonomous organelles like mitochondria, and derive from a cyanobacterial ancestor that was engulfed by a host cell. During evolution, they have recruited proteins originating from the nuclear genome, and only parts of their ancestral metabolic properties were conserved and optimized to limit functional redundancy with other cell c...
Chapter
Chloroplasts are semiautonomous organelles found in plants and protists. They are surrounded by a double membrane system, or envelope. These envelope membranes contain machineries to import nuclear-encoded proteins, and transporters for ions or metabolites, but are also essential for a range of plastid-specific metabolisms. The inner membrane surro...
Article
Full-text available
Context: Higher plants, as autotrophic organisms, are effective sources of molecules. They hold great promise for metabolic engineering, but the behavior of plant metabolism at the network level is still incompletely described. Although structural models (stoichiometry matrices) and pathway databases are extremely useful, they cannot describe the...
Article
Full-text available
Enzymatic and non-enzymatic peroxidation of polyunsaturated fatty acids give rise to accumulation of aldehydes, ketones, and α,β-unsaturated carbonyls of various lengths, known as oxylipins. Oxylipins with α,β-unsaturated carbonyls are reactive electrophile species and are toxic. Cells have evolved several mechanisms to scavenge reactive electrophi...
Data
Superimposition of apo-ceQORH (green), ceQORH-13-KOTE (cyan), and ceQORH-NADP+-13-KOTE (magenta) displaying conformational changes which occur upon binding of 13-KOTE and 13-KOTE + NADP+.
Data
Skeletal formulas of γ-ketol 18:1, 13-KOTE, 13-KODE and drawing of the reaction catalyzed by ceQORH.
Data
Sequence alignment between ceQORH from Arabidopsis thaliana, and QORs from Coxiella burnetii (3TQH) (Franklin et al., 2015), Escherichia coli (1QOR) (Thorn et al., 1995), and Saccharomyces cerevisiae (3QWA) Guo et al., 2011. The ceQORH secondary structure is drawn. The β-strands are represented by arrows and the α-helices by curls. The conserved re...
Data
Inhibition of ceQORH by the ketodiene 13-KODE; (A) activity (ΔAbs340 nm/s) was measured after different preincubation time of the enzyme with 13-KODE. 50 nM enzyme was pre-incubated in the presence of 10 mM HEPES-KOH pH 7.5, 150 mM KCl, 160 μM NADPH and 55 μM 13-KODE. The reaction was initiated by the addition of 25 μM trans-1,3 diphenyl-2-propenon...
Data
Sequence alignment between ceQORH, AtAER (Mano et al., 2002), AtAOR from Arabidopsis thaliana (Yamauchi et al., 2012), and the enone oxidoreductase from Fragaria x ananassa (4IDF) (Schiefner et al., 2013). The conserved residues are highlighted in red. The sequences were aligned using Multalin (Corpet, 1988) and the drawing was generated using ESPr...
Article
Full-text available
Background CaM-like proteins (CMLs) are localized in the cytosol and others in organelles such as the mitochondria, the peroxisomes and the vacuole. To date, although several plastidial proteins were identified as CaM/CML interactors, no CMLs were assigned to the chloroplast. Absence of clues about the genetic identity of plastidial CMLs prevents i...
Article
Full-text available
Copper is a crucial ion in cells, but needs to be closely controlled due to its toxic potential and ability to catalyse the formation of radicals. In chloroplasts, an important step for the proper functioning of the photosynthetic electron transfer chain is the delivery of copper to plastocyanin in the thylakoid lumen. The main route for copper tra...
Data
Electrostatic potential surfaces of P1B-type ATPases. Red denotes negative and blue positive charges, respectively. All structures are limited to the N-domains and oriented the same way, with the nucleotide binding site in the centre and the nucleotide, if present, shown as sticks. For each structure the pdb code is given. In summary the first stru...
Article
Full-text available
Copper is an essential transition metal for living organisms. In the plant model Arabidopsis thaliana, half of the copper content is localized in the chloroplast and, as a cofactor of plastocyanin, copper is essential for photosynthesis. Within the chloroplast, copper delivery to plastocyanin involves two transporters of the PIB-1-ATPases subfamily...
Article
Due to their unique properties, expression and study of membrane proteins in heterologous systems remains difficult. Among the bacterial systems available, the Gram-positive lactic bacterium, Lactococcus lactis, traditionally used in food fermentations, is nowadays widely used for large-scale production and functional characterization of bacterial...
Article
Full-text available
The resurrection plant Haberlea rhodopensis was used to study dynamics of drought response of photosynthetic machinery parallel with changes in primary metabolism. A relation between leaf water content and photosynthetic performance was established, enabling us to perform a non-destructive evaluation of the plant water status during stress. Spectro...
Article
Full-text available
Copper (Cu) plays a key role in the photosynthetic process as cofactor of the plastocyanin (PC), an essential component of the chloroplast photosynthetic electron transfer chain. Encoded by the nuclear genome, PC is translocated in its apo-form into the chloroplast and the lumen of thylakoids where it is processed to its mature form and acquires Cu...
Article
Full-text available
Quinone oxidoreductases reduce a broad range of quinones and are widely distributed among living organisms. The chloroplast envelope quinone oxidoreductase homologue (ceQORH) from Arabidopsis thaliana binds NADPH, lacks a classical N-terminal and cleavable chloroplast transit peptide, and is transported through the chloroplast envelope membrane by...
Article
The study of most membrane proteins remains challenging due to their hydrophobicity and their low natural abundance in cells. Lactococcus lactis, a Gram-positive lactic bacterium, has been traditionally used in food fermentations and is nowadays widely used in biotechnology for large-scale production of heterologous proteins. This system has been s...
Chapter
Full-text available
Membrane proteins (MPs) play key roles in most important cellular processes ranging from cell-to-cell communication to signaling processes. Despite recent improvements, the expression of functionally folded MPs in sufficient amounts for functional and structural characterization remains a challenge. Indeed, it is still difficult to predict whether...
Article
Full-text available
Photosynthesis has shaped atmospheric and ocean chemistries and probably changed the climate as well, as oxygen is released from water as part of the photosynthetic process. In photosynthetic eukaryotes, this process occurs in the chloroplast, an organelle containing the most abundant biological membrane, the thylakoids. The thylakoids of plants an...
Article
Glycerolipids constituting the matrix of photosynthetic membranes, from cyanobacteria to chloroplasts of eukaryotic cells, comprise monogalactosyldiacylglycerol, digalactosyldiacylglycerol, sulfoquinovosyldiacylglycerol and phosphatidylglycerol. This review covers our current knowledge on the structural and functional features of these lipids in va...
Article
Full-text available
Copper is an essential micronutrient but it is also potentially toxic as copper ions can catalyse the production of free radicals, which result in various types of cell damage. Therefore, copper homeostasis in plant and animal cells must be tightly controlled. In the chloroplast, copper import is mediated by a chloroplast-envelope PIB-type ATPase,...
Article
Full-text available
Ca(2+)/Calmodulin (CaM)-dependent signaling pathways play a major role in the modulation of cell responses in eukaryotes. In the chloroplast, few proteins such as the NAD(+) kinase 2 have been previously shown to interact with CaM, but a general picture of the role of Ca(2+)/CaM signaling in this organelle is still lacking. Using CaM-affinity chrom...
Article
Full-text available
Haberlea rhodopensis (Gesneriacea) belongs to the group of the resurrection plants. Their ability to survive extreme desiccation renders them excellent models to study plant response to dehydration. Plant metabolic networks are very complex and subcellular proteomics has proven to be a powerful approach to gain functional knowledge about the sub-ce...
Article
Full-text available
AT_CHLORO (www.grenoble.prabi.fr/at_chloro) is a database dedicated to sub-plastidial localization of A. thaliana chloroplast proteins. This information was infered from proteomics experiments obtained from a comprehensive study that allowed the identification of proteins from envelope, stroma, and thylakoid sub-compartments Ferro et al., 2010. In...
Article
Full-text available
The unicellular green alga Chlamydomonas reinhardtii is a prime model for deciphering processes occurring in the intracellular compartments of the photosynthetic cell. Organelle-specific proteomic studies have started to delineate its various subproteomes, but sequence-based prediction software is necessary to assign proteins subcellular localizati...
Article
Full-text available
Chloroplast ATP/ADP transporters are essential to energy homeostasis in plant cells. However, their molecular mechanism remains poorly understood, primarily due to the difficulty of producing and purifying functional recombinant forms of these transporters. In this work, we describe an expression and purification protocol providing good yields and...
Data
Oligonucleotide sequences of the different primers used for cloning. (DOC)
Chapter
Plastids fulfill a number of essential functions, including photosynthesis, assimilation of nitrogen and sulfur, synthesis of amino acids, fatty acids, and many secondary metabolites. Pure computation-based predictions are limited in predicting plastid proteomes, and proteomic and especially subcellular proteomics studies are essential to provide a...
Article
Full-text available
To study chloroplast metabolism and functions, subplastidial localization is a prerequisite to achieve protein functional characterization. As the accurate localization of many chloroplast proteins often remains hypothetical, we set up a proteomics strategy in order to assign the accurate subplastidial localization. A comprehensive study of Arabido...
Article
Full-text available
Membrane proteins are the targets of 50% of drugs, although they only represent 1% of total cellular proteins. The first major bottleneck on the route to their functional and structural characterisation is their overexpression; and simply choosing the right system can involve many months of trial and error. This work is intended as a guide to where...
Article
Organelle proteomics describes the study of proteins present in organelle at a particular instance during the whole period of their life cycle in a cell. Organelles are specialized membrane bound structures within a cell that function by interacting with cytosolic and luminal soluble proteins making the protein composition of each organelle dynamic...
Article
Full-text available
Copper is an essential plant micronutrient playing key roles in cellular processes, among them photosynthesis. In Arabidopsis thaliana, copper delivery to chloroplasts, mainly studied by genetic approaches, is thought to involve two P(IB)-type ATPases: AtHMA1 and AtHMA6/PAA1. The lack of biochemical characterization of AtHMA1 and PAA1, and more gen...
Article
Eukaryotic membrane protein expression is still a major bottleneck for structural studies. Production in E. coli often leads to low expression level and/or aggregated proteins. In the last decade, strategies relying on new fusion protein expression revealed promising results. Fusion with the amphipatic Mistic protein has been described to favor exp...
Article
Plastids are semiautonomous organelles restricted to plants and protists. These plastids are surrounded by a double membrane system, or envelope. These envelope membranes contain machineries to import nuclear-encoded proteins, and transporters for ions or metabolites, but are also essential for a range of plastid-specific metabolisms. Targeted semi...
Book
Plastids are semiautonomous organdies restricted to plants and protists. These plastids are surrounded by a double membrane system, or envelope. These envelope membranes contain machineries to import nuclear-encoded proteins, and transporters for ions or metabolites, but are also essential for a range of plastid-specific metabolisms. Targeted semiq...
Article
Full-text available
Proteomics has become a critical tool in the functional understanding of plant processes at the molecular level. Proteomics-based studies have also contributed to the ever-expanding array of data in modern biology, with many generating Web portals and online resources that contain incrementally expanding and updated information. Many of these resou...
Article
Recent advances in the proteomic field have allowed high throughput experiments to be conducted on chloroplast samples and the data are available in several databases such as the Plant Protein Database (PPDB), or the SubCellular Proteomic Database (SUBA). However, the accurate localization of many proteins that were identified in different subplast...
Data
Quantification of the recombinant ceQORH protein in L. lactis membranes. L. lactis membrane proteins (MP, 5 and 10 µg) and various amounts of the purified recombinant ceQORH protein produced in E. coli (ceQORHrec, 0.3; 0.6; 1; 2; 3; 4 and 6 µg) were loaded on a 12% SDS-PAGE. Proteins were detected by Coomassie blue staining (upper panel, A) and the...
Data
ATPase activity of control (vv) L. lactis membranes or of L. lactis membranes containing HMA1 or HMA6. The ATPase assay mixture contained 250 mM Tris, pH 7, 15 mM MgSO4, 15 mM ATP, 20 mM Cysteine, 100 µM TCEP, 2 µM CuSO4, and 2 µg of membrane proteins. ATPase activity was followed for 10 min at 37°C. Released inorganic phosphate was colorimetricall...
Data
Chromatographic separation of the ceQORH protein expressed in L. lactis. The recombinant ceQORH protein was first extracted from L. lactis membranes using a salt treatment (1 M NaCl) and then further purified using an affinity chromatography. The purified recombinant ceQORH protein (1.5 mg) was then loaded onto a Superdex 200 10/300 GL column (Phar...
Data
Analysis by SDS-PAGE of recombinant HisceQORH and Strep-tagged ceQORH proteins. A. Analysis by Coomassie blue-stained SDS-PAGE of the recombinant HisceQORH protein produced in L. lactis and E. coli. Increasing amounts of the two recombinant proteins (concentration range: 3, 6 and 9 µg) were loaded on the same gel. Note that the higher proportion of...
Article
Full-text available
Despite their functional and biotechnological importance, the study of membrane proteins remains difficult due to their hydrophobicity and their low natural abundance in cells. Furthermore, into established heterologous systems, these proteins are frequently only produced at very low levels, toxic and mis- or unfolded. Lactococcus lactis, a gram-po...