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July 2, 10:28 AM
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Nuclear receptor signaling via NHR-49/MDT-15 regulates stress resilience and proteostasis in response to reproductive and metabolic cues

Nuclear receptor signaling via NHR-49/MDT-15 regulates stress resilience and proteostasis in response to reproductive and metabolic cues | I2BC Paris-Saclay | Scoop.it

The production of unhealthy eggs leads to improved maternal stress resilience and proteostasis in C. elegans. This study identifies the nuclear hormone receptor NHR-49 as a critical regulator that is activated by elevated fat stores and acts by potentiating the heat shock response.

The ability to sense and respond to proteotoxic insults declines with age, leaving cells vulnerable to chronic and acute stressors. Reproductive cues modulate this decline in cellular proteostasis to influence organismal stress resilience in Caenorhabditis elegans We previously uncovered a pathway that links the integrity of developing embryos to somatic health in reproductive adults. Here, we show that the nuclear receptor NHR-49, an ortholog of mammalian peroxisome proliferator-activated receptor α (PPARα), regulates stress resilience and proteostasis downstream from embryo integrity and other pathways that influence lipid homeostasis and upstream of HSF-1. Disruption of the vitelline layer of the embryo envelope, which activates a proteostasis-enhancing intertissue pathway in somatic cells, triggers changes in lipid catabolism gene expression that are accompanied by an increase in fat stores. NHR-49, together with its coactivator, MDT-15, contributes to this remodeling of lipid metabolism and is also important for the elevated stress resilience mediated by inhibition of the embryonic vitelline layer. Our findings indicate that NHR-49 also contributes to stress resilience in other pathways known to change lipid homeostasis, including reduced insulin-like signaling and fasting, and that increased NHR-49 activity is sufficient to improve proteostasis and stress resilience in an HSF-1-dependent manner. Together, our results establish NHR-49 as a key regulator that links lipid homeostasis and cellular resilience to proteotoxic stress.

More information: https://genesdev.cshlp.org/content/early/2024/05/30/gad.351829.124

Contact: Ambre Sala  ambre.sala@i2bc.paris-saaclay.fr

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April 15, 9:18 AM
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Students from CentraleSupélec visit I2BC groups of the Cellular Biology department

Students from CentraleSupélec visit I2BC groups of the Cellular Biology department | I2BC Paris-Saclay | Scoop.it

80 students of the Bachelor of Global Engineering, a joint novel program of CentraleSupéléc and McGilles University visited the research groups Kühl, Sala and Siudeja at the Cellular Biology department at I2BC in March.

During two afternoons in March three groups from the Cellular Biology department, Kühl, Sala and Siudeja, explained their research questions and model organisms to a total of 80 students from the Bachelor of Global Engineering (BoGe), a joint novel program of CentraleSupéléc, Paris-Saclay and McGilles University, Canada (https://www.centralesupelec.fr/en/bachelor-global-engineering) . The visit was part of the BoGe Biology course taught by Sébastien Bloyer (Genome dep.) and Inge Kühl (BioCell dep.). We thank all the participants for their help, and the I2BC for the support.

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April 15, 8:35 AM
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A Fatty Acid Anabolic Pathway in Specialized-Cells Sustains a Remote Signal that Controls Egg Activation in Drosophila

A Fatty Acid Anabolic Pathway in Specialized-Cells Sustains a Remote Signal that Controls Egg Activation in Drosophila | I2BC Paris-Saclay | Scoop.it

While egg activation, i.e. oocyte to embryo transition, was not known to depend on physiological non-genital signal, this study shows that in Drosophila it depends on a very-long-chain-fatty-acid produced in the oenocytes (lipid metabolism specialized cells). 

Egg activation, representing the critical oocyte-to-embryo transition, provokes meiosis completion, modification of the vitelline membrane to prevent polyspermy, and translation of maternally provided mRNAs. This transition is triggered by a calcium signal induced by spermatozoon fertilization in most animal species, but not in insects. In Drosophila melanogaster, mature oocytes remain arrested at metaphase-I of meiosis and the calcium-dependent activation occurs while the oocyte moves through the genital tract. Here, we discovered that the oenocytes of fruitfly females are required for egg activation. Oenocytes, cells specialized in lipid-metabolism, are located beneath the abdominal cuticle. In adult flies, they synthesize the fatty acids (FAs) that are the precursors of cuticular hydrocarbons (CHCs), including pheromones. The oenocyte-targeted knockdown of a set of FA-anabolic enzymes, involved in very-long-chain fatty acid (VLCFA) synthesis, leads to a defect in egg activation. Given that some but not all of the identified enzymes are required for CHC/pheromone biogenesis, this putative VLCFA-dependent remote control may rely on an as-yet unidentified CHC or may function in parallel to CHC biogenesis. Additionally, we discovered that the most posterior ventral oenocyte cluster is in close proximity to the uterus. Since oocytes dissected from females deficient in this FA-anabolic pathway can be activated in vitro, this regulatory loop likely operates upstream of the calcium trigger. To our knowledge, our findings provide the first evidence that a physiological extra-genital signal remotely controls egg activation. Moreover, our study highlights a potential metabolic link between pheromone-mediated partner recognition and egg activation.

More information: https://doi.org/10.1371/journal.pgen.1011186

Contact: Jacques Montagne jacques.montagne@i2bc.paris-saclay.fr

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January 30, 9:23 AM
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Toxoplasma membrane inositol phospholipid binding protein TgREMIND is essential for secretory organelle function and host infection

Toxoplasma membrane inositol phospholipid binding protein TgREMIND is essential for secretory organelle function and host infection | I2BC Paris-Saclay | Scoop.it

Apicomplexan parasites possess specialized secretory organelles called rhoptries, micronemes, and dense granules that play a vital role in host infection. In this study, we demonstrate that TgREMIND, a protein found in Toxoplasma gondii, is necessary for the biogenesis of rhoptries and dense granules. TgREMIND contains a Fes-CIP4 homology-Bin/Amphiphysin/Rvs (F-BAR) domain, which binds to membrane phospholipids, as well as a novel uncharacterized domain that we have named REMIND (regulator of membrane-interacting domain). Both the F-BAR domain and the REMIND are crucial for TgREMIND functions. When TgREMIND is depleted, there is a significant decrease in the abundance of dense granules and abnormal transparency of rhoptries, leading to a reduction in protein secretion from these organelles. The absence of TgREMIND inhibits host invasion and parasite dissemination, demonstrating that TgREMIND is essential for the proper function of critical secretory organelles required for successful infection by Toxoplasma.

More information: doi: 10.1016/j.celrep.2023.113601

Contact: Stanislas TOMAVO <stanislas.tomavo@i2bc.paris-saclay.fr>

 
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December 27, 2023 9:21 AM
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Visualization and Quantification of Endogenous Intra-Organelle Protein Interactions at ER-Mitochondria Contact Sites by Proximity Ligation Assays

Visualization and Quantification of Endogenous Intra-Organelle Protein Interactions at ER-Mitochondria Contact Sites by Proximity Ligation Assays | I2BC Paris-Saclay | Scoop.it

Meet me at MAM: optimization of a proximity ligation-3D image analysis protocol to quantify protein interactions at Mitochondria-Associated Endoplasmic Reticulum Membranes.

More information:  https://www.jove.com/fr/v/64750/visualization-quantification-endogenous-intra-organelle-protein

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July 19, 2023 10:26 AM
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LGG-1/GABARAP lipidation is not required for autophagy and development in Caenorhabditis elegans

LGG-1/GABARAP lipidation is not required for autophagy and development in Caenorhabditis elegans | I2BC Paris-Saclay | Scoop.it

Using a CRISPR approach in C. elegans, this study shows that the ubiquitin-like ATG8 homolog LGG-1 possesses lipidation-independent function in autophagy, and provides a novel insight into the role of ATG family during animal development.

The ubiquitin-like proteins Atg8/LC3/GABARAP are required for multiple steps of autophagy, such as initiation, cargo recognition and engulfment, vesicle closure and degradation. Most of LC3/GABARAP functions are considered dependent on their post-translational modifications and their association with the autophagosome membrane through a conjugation to a lipid, the phosphatidyl-ethanolamine. Contrarily to mammals, C. elegans possesses single homologs of LC3 and GABARAP families, named LGG-2 and LGG-1. Using site-directed mutagenesis, we inhibited the conjugation of LGG-1 to the autophagosome membrane and generated mutants that express only cytosolic forms, either the precursor or the cleaved protein. LGG-1 is an essential gene for autophagy and development in C. elegans, but we discovered that its functions could be fully achieved independently of its localization to the membrane. This study reveals an essential role for the cleaved form of LGG-1 in autophagy but also in an autophagy-independent embryonic function. Our data question the use of lipidated GABARAP/LC3 as the main marker of autophagic flux and highlight the high plasticity of autophagy.

More information: https://elifesciences.org/articles/85748

Contact: Renaud Legouis  <renaud.legouis@i2bc.paris-saclay.fr>

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March 17, 2023 11:54 AM
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The luminal domain of Toxoplasma gondii sortilin adopts a ring-shaped structure exhibiting motifs specific to apicomplexan parasites

The luminal domain of Toxoplasma gondii sortilin adopts a ring-shaped structure exhibiting motifs specific to apicomplexan parasites | I2BC Paris-Saclay | Scoop.it

Rhoptries and micronemes are essential for host cell invasion and survival of all apicomplexan parasites, which are composed of protozoan pathogens including Plasmodium falciparum (malaria) and Toxoplasma gondii (toxoplasmosis) that infect humans and animals causing severe diseases. We identifiedToxoplasma gondii TgSORT as an essential cargo receptor, which drives the transport of rhoptry and microneme proteins to ensure the biogenesis of these secretory organelles. Here, we present an optimized protocol for expression of the entire luminal N-terminus of TgSORT (Tg-NSORT) in the yeast Pichia pastoris. Optimization of its coding sequence, cloning and transformation of the yeast P. pastoris allowed the secretion of Tg-NSORT. The protein was purified and further analyzed by negative staining electron microscopy. In addition, molecular modeling using AlphaFold identified key differences between the human and theT gondii sortilin. The structural features that are only present inT. gondii and other apicomplexan parasites were highlighted. Elucidating the roles of these specific structural features may be useful for designing new therapeutic agents against apicomplexan parasites.

More: https://doi.org/10.3389/fpara.2023.1103772

Contact: Stanislas Tomavo <stanislas.tomavo@i2bc.paris-saclay.fr>

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December 2, 2022 10:58 AM
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Proton exchange by the vacuolar nitrate transporter CLCa is required for plant growth and nitrogen use efficiency

Proton exchange by the vacuolar nitrate transporter CLCa is required for plant growth and nitrogen use efficiency | I2BC Paris-Saclay | Scoop.it

CLCa exchange mechanism limits nitrogen use efficicency in plants.

Nitrogen (N) is quantitatively the most important inorganic nutrient for plants. Roots mainly take it up in the form of nitrate (NO3-). Once inside the cells, nitrate can be assimilated to amino acids or stored in the vacuole, where it regulates cell water content to sustain plant growth. CLCa is the main transporter mediating the entry of NO3- into vacuoles. It works as an exchanger removing one proton from the vacuole to store two NO3- anions. CLCa belongs to a conserved family composed of both anions/protons exchangers and anions channels, with closely similar structures. Most of the exchangers share a conserved glutamate residue (E203 in CLCa). In this publication, we analyzed if the NO3-/H+ exchanger mechanism of CLCa is required for plants to stabilize water and nitrate status and what are the physiological consequences of the conversion of CLCa from an exchanger to a nitrate channel. We generated Arabidopsis lines that express a mutated form of CLCa in the amino acid E203, turning this protein into a NO3- channel. This modification decreases nitrate accumulation in the vacuole and increases amino acids and protein synthesis, thereby leading to high N content in the seeds. Although these plants present a higher nitrogen use efficiency (NUE), their growth is reduced, in association to a diminution of water content. This finding reveals the importance of the CLCa exchanger mechanism in allowing plant cells to maintain cell turgor irrespective of fluctuating nitrogen concentrations in the soil.

More information: here

Contact: Sophie Filleur <sophie.filleur@i2bc.paris-saclay.fr>

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October 24, 2022 5:09 AM
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ORP5 AND ORP8 ORCHESTRATE LIPID DROPLET BIOGENESIS AT ENDOPLASMIC RETICULUM-MITOCHONDRIA CONTACT SITES

ORP5 AND ORP8 ORCHESTRATE LIPID DROPLET BIOGENESIS AT ENDOPLASMIC RETICULUM-MITOCHONDRIA CONTACT SITES | I2BC Paris-Saclay | Scoop.it

It takes two to birth a lipid droplet in the cell: the encounter of endoplasmic reticulum with mitochondria membranes regulates lipid droplet biogenesis in ORP5/8-dependent way.

Lipid droplets (LDs) are intracellular organelles that play a central role in lipid metabolism and regulate cellular energy balance. In their hydrophobic core, surrounded by a phospholipid monolayer, LDs store energy in the form of neutral lipids and prevent their toxic accumulation in the cell. LDs are formed from the endoplasmic reticulum (ER) membrane and alterations in their biogenesis are associated with many metabolic diseases such as diabetes and heart disease or viral infections. However, the molecular mechanisms that spatially and temporally regulate LD biogenesis in the cell remain largely unknown.
The teams of Francesca Giordano at the I2BC (https://www.i2bc.paris-saclay.fr/lipid-trafficking-and-membrane-contact-sites/) and Abdou Rachid Thiam at the ENS (https://arthiam.com), and their close collaborators such as the ImagerieGif platform at I2BC, have addressed this question using complementary cell biology and imaging approaches. Their study, recently published in Journal of Cell Biology (https://doi.org/10.1083/jcb.202112107) reveals a key role of the lipid transfer proteins ORP5 and ORP8 in the regulation of LD biogenesis at the mitochondria-associated membrane subdomains of the ER (MAM). ORP5/ORP8 control the recruitment to MAM of seipin, an ER protein critical for early steps of LD biogenesis, and their loss or inhibition impairs LD biogenesis at ER-mitochondria contact sites.
This study highlights a key role of ORP5/ORP8 proteins in orchestrating LD biogenesis at MAM and provides new mechanistic insights into the metabolic crosstalk between mitochondria, ER and LDs at membrane contact sites.

More information: https://doi.org/10.1083/jcb.202112107

Contact: Francesca Giordano  <francesca.giordano@i2bc.paris-saclay.fr>

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September 21, 2022 11:35 AM
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E-cadherin acts as a positive regulator of the JAK-STAT signaling pathway during Drosophila oogenesis

E-cadherin acts as a positive regulator of the JAK-STAT signaling pathway during Drosophila oogenesis | I2BC Paris-Saclay | Scoop.it

What is Drosophila teaching us? Usually famous for being part of epithelial adherens junctions, the E-Cadherin conserved protein is identified in this work as a new regulator of the famous JAK-STAT signaling pathway.

The JAK-STAT pathway is evolutionary conserved. The simplicity of this signaling in Drosophila, due to the limited redundancy between pathway components, makes it an ideal model for investigation. In the Drosophila follicular epithelium, highly stereotyped functions of JAK-STAT signaling have been well characterized, but how signaling activity is regulated precisely to allow the different outcomes is not well understood. In this tissue, the ligand is secreted by the polar cells positioned at each follicle extremity, thus generating a gradient of JAK-STAT activity in adjacent cells. One way to control the delivered quantity of ligand is by regulating the number of polar cells, which is reduced by apoptosis to exactly two at each pole by mid-oogenesis. Hence, JAK-STAT activity is described as symmetrical between follicle anterior and posterior regions. Here, we show that JAK-STAT signaling activity is actually highly dynamic, resulting in asymmetry between poles by mid-oogenesis. Interestingly, we found similar temporal dynamics at follicle poles in the accumulation of the adherens junction E-cadherin protein. Remarkably, E-cadherin and JAK-STAT signaling not only display patterning overlaps but also share functions during oogenesis. In particular, we show that E-cadherin, like JAK-STAT signaling, regulates polar cell apoptosis non-cell-autonomously from follicle cells. Finally, our work reveals that E-cadherin is required for optimal JAK-STAT activity throughout oogenesis and that E-cadherin and Stat92E, the transcription factor of the pathway, form part of a physical complex in follicle cells. Taken together, our study establishes E-cadherin as a new positive regulator of JAK-STAT signaling during oogenesis. Fig legend: A Drosophila ovarian follicle stained for the adherens junction marker E-Cadherin showing the honeycomb-like structure of the epithelium.

More information here

Contact: Marianne Malartre et Anne-Marie Pret  <marianne.malartre@i2bc.paris-saclay.fr> et<anne-marie.pret@i2bc.paris-saclay.fr>

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Plant mechanotransduction in the spotlight of Mechano-Sensitive channels

Plant mechanotransduction in the spotlight of Mechano-Sensitive channels | I2BC Paris-Saclay | Scoop.it

Mechanosensitive channels provide to the plant microprobe which convert instantaneously force into biological signals.

The study of mechanosensitive channels (MS) in living organisms has progressed considerably over the past two decades. The understanding of their roles in mechanosensation and mechanotransduction was consecrated by the awarding the Nobel Prize in 2021 to A. Patapoutian for his discoveries on the role of MS channels in mechanoperception in humans. In plant, identified MS channels belong to MSL, MCA, Piezo, OSCA and TPK families. They convert instantaneously (ms range) membrane tension variations into biological signals. Upon activation, MS channels mediate Ca2+ signals, electrical signals or osmotic signals. The next challenge in plant mechanotransduction is to map forces at the cellular scale in order understand under which conditions and at which locations MS channels activate “in cellulo”. This article was published in Current Opinion in Plant Biology.

More information: https://authors.elsevier.com/a/1fJjO4tPF3q9Qk

Contact: Jean-Marie FRACHISSE <jean-marie.frachisse@i2bc.paris-saclay.fr>

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Starting the engine of the powerhouse: mitochondrial transcription and beyond

Starting the engine of the powerhouse: mitochondrial transcription and beyond | I2BC Paris-Saclay | Scoop.it

Mitochondria are the powerhouses of cells thanks to the oxidative phosphorylation system partially encoded by a small mitochondrial genome (mtDNA). Miranda et al review our current knowledge on mtDNA expression, associated disease phenotypes, and how mtDNA and nuclear gene expression are coordinated.

Mitochondria are central hubs for cellular metabolism, coordinating a variety of metabolic reactions crucial for human health. Mitochondria provide most of the cellular energy via their oxidative phosphorylation (OXPHOS) system, which requires the coordinated expression of genes encoded by both the nuclear (nDNA) and mitochondrial genomes (mtDNA). Transcription of mtDNA is not only essential for the biogenesis of the OXPHOS system, but also generates RNA primers necessary to initiate mtDNA replication. Like the prokaryotic system, mitochondria have no membrane-based compartmentalization to separate the different steps of mtDNA maintenance and expression and depend entirely on nDNA-encoded factors imported into the organelle. Our understanding of mitochondrial transcription in mammalian cells has largely progressed, but the mechanisms regulating mtDNA gene expression are still poorly understood despite their profound importance for human disease. Here, we review mechanisms of mitochondrial gene expression with a focus on the recent findings in the field of mammalian mtDNA transcription and disease phenotypes caused by defects in proteins involved in this process.

More information: https://www.degruyter.com/document/doi/10.1515/hsz-2021-0416/html

Contact: Inge Kuhl  <inge.kuhl@i2bc.paris-saclay.fr>

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Scientific day in memory of Agnès Delahodde

Scientific day in memory of Agnès Delahodde | I2BC Paris-Saclay | Scoop.it
Throughout the day, there will be scientific presentations by colleagues and collaborators and personal tributes to Agnès Delahodde.

 

Josette Banroques - IBPC, Paris & Bruno Sargueil - Faculty of Pharmacy, Paris

DNA endonuclease and RNA maturase activities of two intron-encoded proteins from yeast mitochondria

Teresa Rinaldi - Universita la Sapienza, Rome

Mitochondrial function and lipids in S. cerevisiae

Monique Bolotin - GQE-Le Moulon, Gif-sur-Yvette

Vincent Procaccio - CHU, Angers

Disease modeling and drug screening of mitochondrial complex I disorders: from Podospora anserina to Human

Agnès Rötig - Institut Imagine, Paris & Véronique Paquis - CHU, Nice

Yeast does not lose the north or the south in the field of mitochondrial diseases:

 in the North Agnès Rötig

 in the South Véronique Paquis

Laras Pitayu - IJM, Paris

(Mitochondrial) Genome Stability Testimony

Renaud Legouis - I2BC, Gif-sur-Yvette

Mitochondrial rebuilding after acute heat stress relies on autophagy

 

Free but required registration before 22 April 2022

https://evento.renater.fr/survey/inscriptionjournee-d-hommage-agnes-delahodde-ideapch1

Capacity 200 people + Video broadcast

more information: here

Contact: day4agnes@i2bc.paris-saclay.fr

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“Ménage à trois”: emerging function of the mitochondria-ER-lipid droplet connection in coordinating lipid transfer, metabolism and storage in cells

“Ménage à trois”: emerging function of the mitochondria-ER-lipid droplet connection in coordinating lipid transfer, metabolism and storage in cells | I2BC Paris-Saclay | Scoop.it

“Ménage à trois”: coordination of lipid transfer, metabolism and storage at the new mitochondria-ER-lipid droplet junction.

Over the past two decades, we have witnessed a growing appreciation for the importance of membrane contact sites (CS) in facilitating direct communication between organelles. CS are tiny regions where the membranes of two organelles meet but do not fuse and allow the transfer of metabolites between organelles, playing crucial roles in the coordination of cellular metabolic activities. The significant advancements in imaging techniques, and molecular and cell biology research has revealed that CS are more complex than what originally thought, as they are extremely dynamics, they can remodel their shape, composition and functions in accord to metabolic and environmental changes, and can occur between more than two organelles. In this review, for the Special Issue “Lipid droplets, metabolic hubs in health and disease“ in FEBS Letters, Vera F. Monteiro-Cardoso and Francesca Giordano describe how recent studies led to the identification of a three-way mitochondria-ER-lipid droplet CS and discuss the emerging functions of these contacts in maintaining lipid storage, homeostasis and balance. They also summarize properties and functions of key protein components localized at the mitochondria-ER-lipid droplet interface, with a special focus on lipid transfer proteins. Understanding tripartite CS is essential for unraveling the complexities of inter-organelle communication and cooperation within cells.

more information: https://febs.onlinelibrary.wiley.com/doi/10.1002/1873-3468.14893

contact: Francesca GIORDANO <francesca.giordano@i2bc.paris-saclay.fr>

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Male manipulation impinges on social-dependent tumor suppression in Drosophila melanogaster females

Male manipulation impinges on social-dependent tumor suppression in Drosophila melanogaster females | I2BC Paris-Saclay | Scoop.it

This study shows that tumorous Drosophila females perceive a cognitive-like signal from their social environment that acts on tumor growth, while this effect is lost in mated females because of peptides transmitted through the male ejaculate.

Physiological status can influence social behavior, which reciprocally can impinge on physiology and health. Previously, we reported that tumor growth in Drosophila virgin females depends on the social context, but did not investigate the underlying physiological mechanisms. Here, we sought to characterize the signal perceived between tumorous flies, ultimately discovering that the tumor suppressive effect varies depending on reproductive status. Firstly, we show that the tumor suppressive effect is neither dependent on remnant pheromone-like products nor on the microbiota. Transcriptome analysis of the heads of these tumorous flies reveals social-dependent gene-expression changes related to nervous-system activity, suggesting that a cognitive-like relay might mediate the tumor suppressive effect. The transcriptome also reveals changes in the expression of genes related to mating behavior. Surprisingly, we observed that this social-dependent tumor-suppressive effect is lost in fertilized females. After mating, Drosophila females change their behavior ─favoring offspring survival─ in response to peptides transferred via the male ejaculate, a pehnomenon called “male manipulation”. Remarkably, the social-dependent tumor suppressive effect is restored in females mated by sex-peptide deficient males. Since male manipulation has likely been selected to favor male gene transmission, our findings indicate that this evolutionary trait impedes social-dependent tumor growth slowdown.

more information: https://doi.org/10.1038/s41598-024-57003-3

contact: Jacques MONTAGNE <jacques.montagne@i2bc.paris-saclay.fr>

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JAK-STAT-dependent contact between follicle cells and the oocyte controls Drosophila anterior-posterior polarity and germline development

JAK-STAT-dependent contact between follicle cells and the oocyte controls Drosophila anterior-posterior polarity and germline development | I2BC Paris-Saclay | Scoop.it

The authors identified a population of somatic cells in Drosophila follicles that elaborate filopodia penetrating the oocyte, thereby maintaining the contact between the two tissues. This is essential to control polarity and germline development of the future embryo.

The number of embryonic primordial germ cells is determined, in Drosophila, by the quantity of germ plasm, whose assembly starts in the posterior region of the oocyte during oogenesis. Here, we report that extending JAK-STAT activity in the posterior somatic follicular epithelium leads to an excess of primordial germ cells in the future embryo. We show that JAK-STAT signaling is necessary for the differentiation of approximately 20 specialized follicle cells maintaining tight contact with the oocyte. These cells define, in the underlying posterior oocyte cortex, the anchoring of the germ cell determinant oskar mRNA. We reveal that the apical surface of these posterior anchoring cells extends long filopodia penetrating the oocyte. We identify two JAK-STAT targets in these cells that are each sufficient to extend the zone of contact with the oocyte, thereby leading to production of extra primordial germ cells. JAK-STAT signaling thus determines a fixed number of posterior anchoring cells required for anterior-posterior oocyte polarity and for the development of the future germline.

More information: https://doi-org.insb.bib.cnrs.fr/10.1038/s41467-024-45963-z

Contact: Marianne MALARTRE <marianne.malartre@i2bc.paris-saclay.fr>

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Schizosaccharomyces pombe as a fundamental model for research on mitochondrial gene expression: Progress, achievements and outlooks

Schizosaccharomyces pombe as a fundamental model for research on mitochondrial gene expression: Progress, achievements and outlooks | I2BC Paris-Saclay | Scoop.it

This is a comprehensive and critical review on mitochondrial gene expression in fission yeast, presenting up-to-date knowledge, and emphasising numerous contributions of the unicellular model to both fundamental and biomedical research.

Schizosaccharomyces pombe (fission yeast) is an attractive model for mitochondrial research. The organism resembles human cells in terms of mitochondrial inheritance, mitochondrial transport, sugar metabolism, mitogenome structure, and dependence of viability on the mitogenome (the petite-negative phenotype). Transcriptions of these genomes produce only a few polycistronic transcripts, which then undergo processing as per the tRNA punctuation model. In general, the machinery for mitochondrial gene expression is structurally and functionally conserved between fission yeast and humans. Furthermore, molecular research on S. pombe is supported by a considerable number of experimental techniques and database resources. Owing to these advantages, fission yeast has significantly contributed to biomedical and fundamental research. Here, we review the current state of knowledge regarding S. pombe mitochondrial gene expression, and emphasise the pertinence of fission yeast as both a model and tool, especially for studies on mitochondrial translation.

More information: https://doi.org/10.1002/iub.2801

Contact: Nathalie BONNEFOY <nathalie.bonnefoy@i2bc.paris-saclay.fr>

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Straining the root on and off triggers local calcium signaling

Straining the root on and off triggers local calcium signaling | I2BC Paris-Saclay | Scoop.it

Combining microfluidics and imaging, reveal that a pressure exerted on the root induces an elastic deformation and a calcium signal both at the onset and at the offset of the stimulation.

 How plant roots perceive mechanical cues encountered in the soil remains elusive. Combining microfluidics and imaging, we have shown that a pressure exerted on the root induces an elastic deformation and a calcium signal both at the onset and at the offset of the stimulation. Although the intensity of the calcium response increased with the pressure applied, successive stimuli led to a remarkable attenuation of the calcium signal. The calcium elevation was restricted to the tissue under pressure without propagation. This study published in Proceedings of the Royal Society B (https://doi.org/10.1098/rspb.2023.1462) contributes to elucidate the mechanisms of root adaptation to the mechanical cues generated by the soil.
This work was supported by the Région Ile de France through the DIM ELICIT program and by Saclay Plant Sciences through the DYNANO project.

More information: doi: 10.1128/msphere.00401-23 

Contact: Jean-Marie Frachisse – Sébastien Thomine <jean-marie.frachisse@i2bc.paris-saclay.fr> <sebastien.thomine@i2bc.paris-saclay.fr>

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An image-based high-content screening for compounds targeting Toxoplasma gondii repurposed inhibitors effective against the malaria parasite Plasmodium falciparum

An image-based high-content screening for compounds targeting Toxoplasma gondii repurposed inhibitors effective against the malaria parasite Plasmodium falciparum | I2BC Paris-Saclay | Scoop.it

Apicomplexan parasites contain specific secretory organelles called rhoptries and micronemes, whose biogenesis depends on the unique receptor sortilin TgSORT. In the present study, we took advantage of the subcellular polarity of the parasite to engineer a clonal transgenic Toxoplasma line that expresses simultaneously the green fluorescent protein TgSORT-GFP in the post-Golgi-endosome-like compartment and the red fluorescent protein rhoptry ROP1-mCherry near the apical end. We utilized this fluorescent transgenic T. gondii to develop a miniaturized image-based phenotype assay coupled to an automated image analysis. By applying this methodology to 1,120 compounds, we identified 12 that are capable of disrupting the T. gondii morphology and inhibiting intracellular replication. Analysis of the selected compounds confirmed that all 12 are kinase inhibitors and intramembrane pumps, with some exhibiting potent activity against Plasmodium falciparum. Our findings highlight the advantage of comparative and targeted phenotypic analysis involving two related parasite species as a means of identifying molecules with a conserved mode of action.

More: https://doi.org/10.3389/fcimb.2023.1102551

Contact: Stanislas Tomavo <stanislas.tomavo@i2bc.paris-saclay.fr>

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Biolistic Transformation of Chlamydomonas reinhardtii and Saccharomyces cerevisiae Mitochondria

Biolistic Transformation of Chlamydomonas reinhardtii and Saccharomyces cerevisiae Mitochondria | I2BC Paris-Saclay | Scoop.it

This review summarizes principles, historical and practical aspects as well as achievements of microalga and budding yeast mitochondrial DNA manipulation using the biolistic technique, and includes a video protocol.

Chlamydomonas reinhardtii and Saccharomyces cerevisiae are currently the two micro-organisms in which genetic transformation of mitochondria is routinely performed. The generation of a large variety of defined alterations as well as the insertion of ectopic genes in the mitochondrial genome (mtDNA) are possible, especially in yeast. Biolistic transformation of mitochondria is achieved through the bombardment of microprojectiles coated with DNA, which can be incorporated into mtDNA thanks to the highly efficient homologous recombination machinery present in S. cerevisiae and C. reinhardtii organelles. Despite a low frequency of transformation, the isolation of transformants in yeast is relatively quick and easy, since several natural or artificial selectable markers are available, while the selection in C. reinhardtii remains long and awaits new markers. Here, we describe the materials and techniques used to perform biolistic transformation, in order to mutagenize endogenous mitochondrial genes or insert novel markers into mtDNA. Although alternative strategies to edit mtDNA are being set up, so far, insertion of ectopic genes relies on the biolistic transformation techniques. This reviews is Chapter 24 of a 2023 volume from Springer Protocols devoted to mitochondrial DNA.

More: Methods Mol Biol 2023;2615:345-364. 

Contcat: Nathalie Bonnefoy <nathalie.bonnefoy@i2bc.paris-saclay.fr>

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A functional protein complex, associated to ciliopathies, dictates the future position of distal appendages

A functional protein complex, associated to ciliopathies, dictates the future position of distal appendages | I2BC Paris-Saclay | Scoop.it

Using BioID and expansion microscopy, the study uncover a conserved functional complex required for determining the future position of centriolar distal appendages and their assembly, allowing the basal body anchoring process during ciliogenesis.

Cilia/flagella play fundamental and diverse biological functions in a wide variety of eukaryotes. The cilia assembly process, also called ciliogenesis, is a multistep process involving 4 major events: basal body (BB) duplication, migration to the cell surface, membrane anchoring of the BB via the distal appendages, and ciliary growth. The conservation of this sequence of events in most phyla is paralleled by an important conservation of the proteins involved. The BB anchoring step requires the tethering of the distal appendages to a membrane. The interaction of the BB with the membrane leads to the formation of the transition zone (TZ), recognized to act as a diffusion barrier between the intracellular space and the cilium. Mutations in numerous genes involved in BB docking and TZ assembly are associated with the most severe ciliopathies highlighting the importance of these events in ciliogenesis.
To discover novel molecular actors involved in the BB anchoring process, the Tassin’s team used BioID technology in human cells. They focused their attention on a protein, CEP90, together with its bait (FOPNL) and another protein (OFD1) involved in ciliopathies and known to interact with FOPNL. These 3 proteins are conserved from Protist to mammals despite absent from some phyla. They unveiled using ultrastructure expansion microscopy that the 3 proteins localize at the distal end of both centrioles/BB in Paramecium and mammalian cells. Functional analysis performed both in Paramecium and mammalian cells demonstrate that the 3 proteins form a functional complex required for distal appendage assembly and BB docking. Intriguingly, these proteins are recruited early during centriole duplication on the external surface of the procentriole. This early recruitment on procentriole led us to propose that this functional complex dictates the future distal appendage location in mammalian cells one or two cell cycle before their assembly.

 

More information here

Contact: Anne-Marie Tassin  <anne-marie.tassin@i2bc.paris-saclay.fr>

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ORP5/8 and MIB/MICOS link ER-mitochondria and intra-mitochondrial contacts for non-vesicular transport of phosphatidylserine

ORP5/8 and MIB/MICOS link ER-mitochondria and intra-mitochondrial contacts for non-vesicular transport of phosphatidylserine | I2BC Paris-Saclay | Scoop.it

The ER lipid transfer proteins ORP5 and ORP8 interact and cooperate with the mitochondrial MIB/MICOS complexes to mediate non-vesicular transport of phosphatidylserine from the ER to the mitochondria.

Mitochondria are dynamic organelles essential for cell survival whose structural and functional integrity rely on selective and regulated transport of lipids from/to the endoplasmic reticulum (ER) and across the mitochondrial intermembrane space. As they are not connected by vesicular transport, the exchange of lipids between ER and mitochondria occurs at membrane contact sites. However, the mechanisms and proteins involved in these processes are only beginning to emerge.
In an article recently published in Cell Reports, the team of Francesca Giordano at the I2BC (Cell Biology Department) and their close collaborators, including the ImagerieGif platform at I2BC, have shown that the lipid transfer proteins ORP5 and ORP8 localize at mitochondria-associated ER membrane (MAM) subdomains physically linked to the MIB/ MICOS complexes that bridge the two mitochondrial membranes. Their study also reveals that ORP5 and ORP8 mediate non-vesicular transport of phosphatidylserine (PS) lipids from the ER to mitochondria by cooperating with the MIB/MICOS complexes. Overall this work uncovers a physical and functional link between ER-mitochondria contacts involved in lipid transfer and intra-mitochondrial membrane contacts maintained by the MIB/MICOS complexes.
On the cover of Cell Reports (September 20, 2022): 3D reconstructions of mitochondrial network and ORP5-ORP8 interactions (proximity ligation assay) in a HeLa cell. Illustration by Vera F. Monteiro-Cardoso.

More information: here

Contact: Francesca Giordano  <francesca.giordano@i2bc.paris-saclay.fr>

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Emergence of a new function after duplication of a metal transporter gene in poplar

Emergence of a new function after duplication of a metal transporter gene in poplar | I2BC Paris-Saclay | Scoop.it

A combination of detailed phylogenetic and functional analyses of a pair of tandem duplicated metal transporter encoding genes uncovers a rare example of neofunctionalization caught in the act.

Gene duplication is an important mechanism in evolution because it opens the possibility to generate proteins with new functions through mutations in one of the copies. However, there are only few examples clearly illustrating this process of neofunctionalization in the literature. Poplar is important economically and as a model tree species. It is also grown in polluted areas in the context of phytoremediation. However, little is known about the mechanisms of metal homeostasis in poplar and in trees in general. The authors have taken advantage of the availability of genomic sequences for a wide array of Populus and Salix species to establish a detailed phylogeny of NRAMP3 gene in Salicaceae. This revealed a duplication of NRAMP3 metal transporter gene coinciding with the emergence of the genus Populus. Synonymous codon analysis pointed at strong purifying selection on each of the copies, suggesting that both copies encode functional proteins. In parallel, the functional analysis of the two copies in yeast and Arabidopsis demonstrated that both copies encode functional metal transporters but that only one retained the integrated function of the Arabidopsis thaliana homologues. Combining cellular imaging and expression in poplar highlighted a new and unexpected function in cell-to-cell transport of manganese for the other copy, associated with a change in subcellular localization. This work provides a rare example of neofunctionalization of a metal transporter encoding gene after a tandem duplication specific of the genus Populus.

More information: https://doi.org/10.1093/molbev/msac129

Contact: Sébastien Thomine <sebastien.thomine@i2bc.paris-saclay.fr>

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Top match between shade and wounding on leaf growth and movement: an easy win for shade?

Top match between shade and wounding on leaf growth and movement: an easy win for shade? | I2BC Paris-Saclay | Scoop.it

Studying the plant growth-defense tradeoff, the authors show that shade suppresses wound-induced leaf repositioning and identify key players in signal integration.

Plants face different types of stressful situations without the ability to relocate to favorable environments. For example, increasing plant density reduces access to sunlight as plants start to shade each other. Foliar shading represents a stress that many plants cope with by changing their morphology. This includes elongation of stem-like structures and repositioning of leaves to favor access to unfiltered sunlight. Plants also defend themselves against various pathogens including herbivores. Defense mechanisms include the production of deterrent chemical and morphological adaptations such as stunted growth and downwards leaf repositioning.
Here we studied the morphological response of plants when simultaneously facing shade and herbivore stress. When facing both stresses petiole growth was intermediate between the shade-enhanced and wound-repressed response. In contrast, the shade cue overrides the wounding cue leading to a similar upwards leaf repositioning in the combined treatments or in the response to shade alone. Using gene expression analyses and genetics we identified two members of the Phytochrome Kinase Substrate family as playing a signal integration role when plants simultaneously faced both stresses. This contributes to our understanding of the mechanisms underlying plant morphological adaptations when facing multiple stresses.
This work was led by Anne-Sophie Fiorucci during her post-doctoral studies in the lab of Christian Fankhauser (University of Lausanne). It sets the basis for her projects as Assistant Professor in the group of Sébastien Thomine at I2BC. In the frame of the ANR JCJC NitReST she got in 2021, she is now focusing on plant adaptations to shade and temperature in relationship with nitrogen nutrition.

More information: https://journals.plos.org/plosgenetics/article?id=10.1371/journal.pgen.1010213

Contact: Anne-Sophie Fiorucci <anne-sophie.fiorucci@i2bc.paris-saclay.fr>

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Micronutrient homeostasis in plants for more sustainable agriculture and healthier human nutrition

Micronutrient homeostasis in plants for more sustainable agriculture and healthier human nutrition | I2BC Paris-Saclay | Scoop.it

In this prospective review, the authors call for awareness of the importance of micronutrients in crop production and highlight the need for further research on this topic to meet the challenges of developing more sustainable and nutritious crops.

Plants need a range of essential micronutrients. Micronutrient deficiencies affect crop yield and quality in many countries. Mild or hidden micronutrient deficiency likely limits crop yields in much wider areas than those where obvious symptoms occur. Animals, including humans, also require micronutrients to be in good health. Insufficient amount and bioavailability of micronutrients in plant-based diets represent a major reason for the high prevalence of micronutrient deficiencies in human populations. In Europe, these deficiencies are a growing concern among pregnant women, children and elderly people, and are prone to become more severe with European diet transition towards vegetarianism. With this review, the authors call for awareness of the importance of micronutrients in crop production. They stress the need for better micronutrient nutrition in human populations, not only in developing but also in developed nations, and describe strategies to identify and characterize new varieties with high micronutrient content. Furthermore, they highlight how adequate nutrition of plants with micronutrients may improve metabolic functions and the capacity of plants to cope with abiotic and biotic constraints. Research in this area is expected to foster the development of sustainable and healthy food crops for human consumption. This review reports part of a prospective on the need for plant improvement in the EU project CropBooster-P (https://www.cropbooster-p.eu/). Its publication received the support of the COST action PLANTMETALS (https://plantmetals.eu/plantmetals-home.html).

more information: https://doi.org/10.1093/jxb/erac014

contact: Sébastien THOMINE <sebastien.thomine@i2bc.paris-saclay.fr>

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