Biology subjects

Ravanel, S.

Publications and source records attributed to Ravanel, S..

12 recordsLinked to original sources

bioRxiv2026

Integrated cellular and molecular responses to uranium chemotoxicity in the metal-tolerant microalga Coelastrella sp. PCV

Understanding the toxicity of hazardous metals in microalgae is critical for environmental risk assessment and sustainable phycoremediation. Metal-tolerant organisms provide powerful models for dissecting the mechanisms that mitigate metal toxicity. Here, we investigated the cellular and molecular responses to uranium (U) chemotoxicity in the metal-tolerant microalga Coelastrella sp. PCV. We used an integrated multi-omics and high-resolution imaging approach, combined with physiological analyses, to elucidate the mechanisms underlying U tolerance in Coelastrella. Using TEM-EDX, U was localized to the cell wall, polyphosphate bodies within acidocalcisomes, and vacuoles. Three-dimensional cell reconstruction and morphometric analysis using FIB-SEM showed that U-challenged cells displayed increased vacuolization, reflecting sequestration of uranyl ions and autophagy-mediated detoxification. Transcriptome responses were rapid and extensive, characterized by repression of cell division and photosynthesis, and pronounced imbalance in protein turnover and trafficking. Uranium also disrupted the homeostasis of essential elements, with marked rewiring of gene networks governing molybdenum, manganese, phosphate, iron and calcium homeostasis, notably affecting transporters and metal-binding proteins. Coelastrella sp. PCV efficiently sequestered U in acidocalcisomes and vacuoles, while rapidly excluding U from the cell. These coordinated detoxification responses are likely mediated by calcium, iron, ABC, and MATE transporters among the strongly deregulated genes under U stress.

plant biology↗

bioRxiv2025

Chloroplast-mitochondria synergy modulates responses to iron limitation in two Thalassiosira diatom species

Iron is naturally present at low levels in modern oceans, but it remains essential for marine life. Ocean-dwelling organisms such as oceanic phytoplankton must therefore adapt to the available levels. Among phytoplankton, diatoms are a highly diverse and successful taxon that includes the Thalassiosira genus. As a group, diatoms contribute around 20% of global primary photosynthetic productivity, they have also developed specific resources allowing them to thrive in low-iron regions. However, the major biological factors underlying their success in these ocean environments remain unknown. Here, we compared two Thalassiosira species: T. oceanica from iron-poor open-ocean; and T. pseudonana, from iron-rich coastal waters. Since iron is essential for both photosynthesis and respiration, we examined the specificities of the bioenergetic machineries in these organisms using a combination of photo-physiological, proteomics, and FIB-SEM methods. We particularly focused on chloroplast-mitochondrial coupling, a mechanism deployed by diatoms to ensure optimal transfer of photosynthetic products to promote cell growth. This study of this mechanism in the context of iron limitation reveals that the two diatoms differentially remodel chloroplast compartments in response to iron limitation. Their tolerance to these conditions is also linked to distinct constitutive mitochondrial architectures.

plant biology↗

bioRxiv2025

Effects of manganese dioxide on macrophages under different exposure schemes

Manganese dioxide is a material that is more and more used in its particular form, for many industrial applications such as chemical catalysis or for batteries. Thus, workers can be exposed to this particulate chemical. It is known that overexposure to manganese leads to a brain disease called manganism. However, manganese is also known to impact the pulmonary function, which is important as pulmonary exposure is of prime importance for workers. We thus investigated the effects of manganese dioxide on macrophages, i.e. the scavenger cells that take particulates in charge in our bodies. To this purpose, we used a combination of proteomic and targeted approaches, in order to obtain a wide view of the cellular responses to manganese dioxide. We also used a repeated exposure mode, in order to better mimic occupational exposure. Our results point out the fact that manganese oxide nanoparticles are rather toxic for macrophages and induce mitochondrial dysfunction, oxidative stress and a pro-inflammatory response. Environmental significanceManganese dioxide is more and more used in batteries, so that workers in batteries factories are exposed to this metallic oxide. As always for particulate materials, macrophages are the first line of defense of the organism. We thus investigated the effects of manganese dioxide on macrophages, using a repeated exposure scheme to mimic occupational exposure, and the effects were documented by a combination of proteomic and targeted approaches. The functional effects include mitochondrial dysfunction, oxidative stress and inflammation.

pharmacology and toxicology↗

bioRxiv2025

The devil lies in the details: small structural and chemical changes in iron oxide pigments largely alter the biological outcomes in macrophages

Because of their technical qualities, such as resistance to fading and to high temperatures, mineral pigments are still widely used nowadays. Among the diversity of mineral pigments, iron oxide pigments represent a widely-used class, because of their technical qualities, diversity of shades (from yellow to red to brown to black) and low toxicity compared to heavy metals-based pigments. However, a low toxicity does not mean the absence of adverse effects. We thus investigated the effects on macrophages of two different subtypes of Pigment Red 101, i.e. hematite, produced by two different processes, namely a wet precipitation process and a calcination process. Macrophages were chosen as a target cell type because they represent the main scavenger cell type that is in charge of handling particulate materials in the body. During this comparison, we realized that the calcined pigment was contaminated from the start by bacterial endotoxins, which induced intense inflammatory responses and biased the comparison. After depyrogenation, the calcined pigment proved to dissolve to a higher extent in macrophages, but to show less important adverse effects (e.g. alteration of the mitochondrial transmembrane potential, oxidative stress and inflammatory responses) than the precipitated pigment in a recovery exposure mode, allowing to investigate delayed effects of the pigments. Thus, despite their identical pigment number, pigments differing in their structure and in their synthesis induce different responses from living cells, even if administrated in equivalent amount. This should be taken into account for some applications, such as tattooing. Moreover, endotoxin contamination should also be checked to increase workers and users safety

pharmacology and toxicology↗

bioRxiv2025

Beyond the ink: cellular and molecular effects of iron-based pigments on macrophages

As ochre, iron oxide is among the most ancient pigments used by mankind for different purposes, including tattooing as demonstrated on tattoed mummies. Iron oxides are still used in tattooing nowadays and especially in dermopigmentation, an area of medical tattoing aiming at restoring the color of skin. This ancient use of iron oxide does not mean that it has no effect on cells, and especially on macrophages, the cells that maintain pigments particles on site in tattoos. We thus investigated in vitro the delayed/sustained effects of iron oxide pigments on macrophages, i.e. the effects occurring a few days after the exposure to pigments, on pigments-loaded macrophages but in a pigment-free medium, mimicking the status of tattooed skin after all the pigment particles have been captured. By combining proteomic and targeted approaches, we determined that red iron oxide (but not black iron oxide) induces perturbations in mitochondria, altering the mitochondrial transmembrane potential. Red iron oxide also induces oxidative stress and the secretion of pro-inflammatory cytokines such as interleukin 6 and tumor necrosis factor. Thus, red iron oxide induces adverse effects on macrophages that may persist over time, owing to its low intracellular dissolution.

immunology↗

bioRxiv2025

Identification of uranyl-binding proteins in Arabidopsis thaliana cells exposed to uranium: Insights from a metalloproteomic analysis and characterization of Glycine-Rich RNA-binding Protein 7 (GRP7)

Uranium (U) is a naturally occurring radionuclide, chemotoxic for living organisms. To identify proteins that could be cellular targets for U in plants, we used metalloproteomic approaches combining column chromatographic fractionation analyses, protein identification by high-resolution mass spectrometry shotgun proteomics, and metal quantification by induced coupled plasma mass spectrometry. We identified 57 candidate proteins for uranyl (U(VI)) binding in cultured Arabidopsis thaliana cells. One of these proteins, the Glycine-Rich RNA-binding Protein 7 (GRP7) is a RNA-binding protein involved in various developmental processes and responses to biotic and abiotic stress. Recombinant GRP7 was purified from overproducing bacteria and subjected to further biochemical characterization. First, we showed that GRP7 binds U(VI) with a 1:2 (protein:metal) stoichiometry in vitro. Next, we analyzed its structural properties by solution-state nuclear magnetic resonance spectroscopy. This allowed us to gain insight into the molecular dynamics of the protein-metal interaction and to identify residues involved in U(VI)-binding in the two sites. Finally, we observed that U(VI) binding interferes with nucleic acid binding to the protein RNA-binding domain, suggesting that U(VI) binding to GRP7 contributes to U toxicity in plants. HIGHLIGHTSO_LI57 candidate uranyl-binding proteins were identified from Arabidopsis cells exposed to uranyl, of which the RNA-binding protein GRP7. C_LIO_LIIn vitro, recombinant GRP7 binds 2 uranyl ions within the RNA recognition motif domain. C_LIO_LIAmino acid residues involved in uranyl binding in both binding sites were identified. C_LIO_LICompetition between specific oligonucleotide and uranyl for binding suggests the implication of GRP7 in uranium toxicity, as a cellular target. C_LI

biochemistry↗

bioRxiv2024

Subcellular ToF-SIMS imaging of the snow algae Sanguina nivaloides by combining high mass and high lateral resolution acquisitions

Time-of-flight secondary ion mass spectrometry (ToF-SIMS) imaging has demonstrated great potential for metabolic imaging, yet achieving sufficiently high lateral and mass resolution to reach the organelle scale remains challenging. We have developed an approach by combining ToF-SIMS imaging acquisitions at high lateral resolution (> 150 nm) and high mass resolution (9,000). The data were then merged and processed using multivariate analysis (MVA), allowing for the precise identification and annotation of 85% of the main contributors to the multivariate analysis components at high lateral resolution. Insights into the electron microscopy sample preparation are provided, especially as we reveal that at least three different osmium-containing complexes can be found depending on the specific chemical environment of organelles. In cells of the snow alga Sanguina nivaloides, living in a natural environment limited in nutrients such as phosphorus (P), we were able to map elements and molecules within their subcellular context, allowing for the molecular fingerprinting of organelles at a resolution of 100 nm, as confirmed by correlative electron microscopy. It was thus possible to highlight that S. nivaloides likely absorbed selectively some inorganic P forms provided by P-rich dust deposited on the snow surface. S. nivaloides cells could maintain phosphorylations in the stroma of the chloroplast, consistently with the preservation of photosynthetic activity. The presented method can thus overcome the current limitations of ToF-SIMS for subcellular imaging and contribute to the understanding of key questions such as P homeostasis and other cell physiological processes.

plant biology↗

bioRxiv2024

Dynamics of the membrane- and cell wall-associated proteome of Arabidopsis thaliana roots in response to uranium stress

Uranium (U) is a non-essential and toxic metal for plants, which have the ability to take up uranyl ions from the soil and preferentially accumulate them in the roots. We showed that the bulk of the radionuclide accumulates in the root insoluble proteome of Arabidopsis plants challenged with U. Therefore, to elucidate new molecular mechanisms related to U stress response and tolerance, we used label-free quantitative proteomics to analyze the dynamics of the root membrane- and cell wall-enriched proteome under U stress. Of the 2,802 proteins identified, 458 showed differential accumulation in response to U. Biological processes affected by U include response to stress, amino acid metabolism, and previously unexplored functions associated with membranes and the cell wall. Indeed, our analysis supports a dynamic and complex reorganization of the cell wall in response to U stress, including lignin and suberin synthesis, pectin modifications, polysaccharide hydrolysis, and Casparian strips formation. Water flux through aquaporins and vesicular trafficking were also significantly perturbed by U stress. Finally, the abundance of metal transporters and iron, calcium, and other metal-binding proteins was affected by U. These proteins may play a role in controlling the fate and toxicity of U in plants.

plant biology↗

bioRxiv2023

Characterization of a uranium-tolerant green microalga of the genus Coelastrella with high potential for the remediation of metal-polluted waters

Uranium (U) pollution of terrestrial and aquatic ecosystems poses a significant threat to the environment and human health because this radionuclide is chemotoxic. Characterization of organisms that tolerate and accumulate U is critical to decipher the mechanisms evolved to cope with the radionuclide and to propose new effective strategies for bioremediation of U-contaminated environments. Here, we isolated a unicellular green microalga of the genus Coelastrella from U-contaminated wastewater. We showed that Coelastrella sp. PCV is much more tolerant to U than Chlamydomonas reinhardtii and Chlorella vulgaris. Coelastrella is able to accumulate U very rapidly, then gradually release it into the medium, behaving as an excluder to limit the toxic effects of U. The ability of Coelastrella to accumulate U is remarkably high, with up to 600 mg U sorbed per g dry biomass. Coelastrella is able to grow and maintain high photosynthesis in natural metal-contaminated waters from a wetland near a reclaimed U mine. Over a single one-week growth cycle, Coelastrella is able to capture 25-55% of U from contaminated waters and demonstrates lipid droplet accumulation. Coelastrella sp. PCV is a very promising microalga for the remediation of polluted waters with valorization of algal biomass that accumulates lipids. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=176 SRC="FIGDIR/small/546994v1_ufig1.gif" ALT="Figure 1"> View larger version (36K): org.highwire.dtl.DTLVardef@1ff356corg.highwire.dtl.DTLVardef@c53d0org.highwire.dtl.DTLVardef@15284eborg.highwire.dtl.DTLVardef@946d49_HPS_FORMAT_FIGEXP M_FIG C_FIG

plant biology↗

bioRxiv2021

Calcium-permeable cation channels are involved in uranium uptake in Arabidopsis thaliana

Uranium (U) is a non-essential and toxic element that is taken up by plants from the environment. The assimilation pathway of U is still unknown in plants and any other organism. In this study, we provide several evidences that U is taken up by the roots of Arabidopsis thaliana through Ca2+-permeable cation channels. First, we showed that deprivation of Arabidopsis plants with calcium induced a 1.5-fold increase in the capacity of roots to accumulate U, suggesting that calcium deficiency promoted the radionuclide import pathway. Second, we showed that external calcium inhibits U accumulation in roots, suggesting a common route for the uptake of both cations. Third, we found that gadolinium, nifedipine and verapamil inhibit the absorption of U, suggesting that different types of Ca2+-permeable channels serve as a route for U uptake. Last, we showed that U bioaccumulation in Arabidopsis mutants deficient for the Ca2+-permeable channels MCA1 and ANN1 was decreased by 40%. This suggests that MCA1 and ANN1 contribute to the absorption of U in different zones and cell layers of the root. Together, our results describe for the first time the involvement of Ca2+-permeable cation channels in the cellular uptake of U.

plant biology↗

bioRxiv2021

Subcellular architecture and metabolic connection in the planktonic photosymbiosis between Collodaria (radiolarians) and their microalgae

Photosymbiosis is widespread and ecologically important in the oceanic plankton but remains poorly studied. Here, we used multimodal subcellular imaging to investigate the photosymbiosis between colonial Collodaria and their microalga dinoflagellate (Brandtodinium) collected in surface seawaters. We showed that this symbiosis is a very dynamic system whereby symbionts interact with different host cells via extracellular vesicles within the "greenhouse-like" colony. 3D electron microscopy revealed that the volume of the photosynthetic apparatus (plastid and pyrenoid) of the microalgae increased in symbiosis compared to free-living while the mitochondria volume was similar. Stable isotope probing coupled with NanoSIMS showed that carbon and nitrogen were assimilated and stored in the symbiotic microalga in starch granules and purine crystals, respectively. Nitrogen was also allocated to the algal nucleus (nucleolus). After 3 hours, low 13C and 15N transfer was detected in the host Golgi. Metal mapping revealed that intracellular iron concentration was similar in free-living and symbiotic microalgae (ca 40 ppm) and two-fold higher in the host, whereas copper concentration increased in symbiotic microalgae (up to 6900 ppm) and was detected in the host cell and extracellular vesicles. Sulfur mapping also pinpointed the importance of this nutrient for the algal metabolism. This study, which revealed subcellular changes of the morphology and nutrient homeostasis in symbiotic microalgae, improves our understanding on the metabolism of this widespread and abundant oceanic symbiosis and paves the way for more studies to investigate the metabolites exchanged.

ecology↗

bioRxiv2019

Evidence for the involvement of protein lysine methylation in the response of sensitive and tolerant Arabidopsis species to cadmium stress

The mechanisms underlying the response and adaptation of plants to excess of trace elements are not fully described. Here, we analyzed the importance of protein lysine methylation for plants to cope with cadmium. We analyzed the effect of cadmium on lysine-methylated proteins and protein lysine methyltransferases (KMTs) in two cadmium-sensitive species, Arabidopsis thaliana and A. lyrata, and in three populations of A. halleri with contrasting cadmium accumulation and tolerance traits. We showed that some proteins are differentially methylated at lysine residues in response to Cd and that a few genes coding KMTs is regulated by cadmium. Also, we showed that nine out of 23 A. thaliana mutants interrupted in KMT genes have a tolerance to cadmium that is significantly different from that of wild-type seedlings. We further characterized two of these mutants, one was knocked-out in the calmodulin lysine methyltransferase gene and displayed increased tolerance to cadmium, the other was interrupted in a KMT gene of unknown function and showed a decreased capacity to cope with cadmium. Together, our results showed that lysine methylation of non-histone proteins is impacted by cadmium and that several methylation events are important for modulating the response of Arabidopsis plants to cadmium stress.

plant biology↗