2020
- Role of a versatile peptide motif controlling Hox nuclear export and autophagy in the Drosophila fat body
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Duffraisse M; PAUL, R; Carnesecchi, J; Hudry, B; Banreti, A; Reboulet, J; Ajuria, L; Lohmann, I; Merabet, S
- J CELL SCI
- Acknowledged platform: Arthrotools
Abstract :
Hox proteins are major regulators of embryonic development, acting in the nucleus to regulate the expression of their numerous downstream target genes. By analyzing deletion forms of the Drosophila Hox protein Ultrabithorax (Ubx), we identified the presence of an unconventional nuclear export signal (NES) that overlaps with a highly conserved motif originally described as mediating the interaction with the PBC proteins, a generic and crucial class of Hox transcriptional cofactors that act in development and cancer. We show that this unconventional NES is involved in the interaction with the major exportin protein CRM1 (also known as Embargoed in flies) in vivo and in vitro We find that this interaction is tightly regulated in the Drosophila fat body to control the autophagy-repressive activity of Ubx during larval development. The role of the PBC interaction motif as part of an unconventional NES was also uncovered in other Drosophila and human Hox proteins, highlighting the evolutionary conservation of this novel function. Together, our results reveal the extreme molecular versatility of a unique short peptide motif for controlling the context-dependent activity of Hox proteins both at transcriptional and non-transcriptional levels.
SEP 2020
DOI : 10.1242/jcs.241943
Pubmed ID : 32878938
- Stiffness measurement is a biomarker of skin ageing in vivo
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Runel, G; Cario, M; Lopez-Ramirez, N; Malbouyres, M; Ruggiero, F; Bernard, L; Puisieux, A; Caramel, J; Chlasta, J; Masse, I
- EXP DERMATOL
- Collaborative platform: PRECI
Abstract :
Human skin is particularly vulnerable to age-related deterioration and undergoes profound structural and functional changes, reflected in the external skin appearance. Skin ageing is characterized by features such as wrinkling or loss of elasticity. Even if research advances have been done concerning the molecular mechanisms that underlie these changes, very few studies have been conducted concerning the structure stiffness of the skin organ as a whole. In this study, we showed, thanks to human skin reconstructs and the Japanese Medaka fish model, that biomechanics is a new biomarker of skin ageing. We revealed that global stiffness measurement by Atomic Force Microscopy, since modulated through ageing in these models, can be a new biomarker of skin ageing, and reflects the profound reorganization of the dermis extracellular matrix, as shown by Transmission Electron Microscopy. Moreover, our data unveiled that the Japanese Medaka fish could represent a highly relevant integrated model to study skin ageing in vivo.
DEC 2020
DOI : 10.1111/exd.14195
Pubmed ID : 32967047
- BMP-1 disrupts cell adhesion and enhances TGF-β activation through cleavage of the matricellular protein thrombospondin-1
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Anastasi, C; Rousselle, P; Talantikite, M; Tessier, A; Cluzel, C; Bachmann, A; Mariano, N; Dussoyer, M; Alcaraz, LB; Fortin, L; Aubert, A; Delolme, F; El Kholti, N; Armengaud, J; Fournié, P; Auxenfans, C; Valcourt, U; Goff, SV; Moali, C
- SCI SIGNAL
- Collaborative platform: PSF; Acknowledged platform: PLATIM, Cytometry
Abstract :
Bone morphogenetic protein 1 (BMP-1) is an important metalloproteinase that synchronizes growth factor activation with extracellular matrix assembly during morphogenesis and tissue repair. The mechanisms by which BMP-1 exerts these effects are highly context dependent. Because BMP-1 overexpression induces marked phenotypic changes in two human cell lines (HT1080 and 293-EBNA cells), we investigated how BMP-1 simultaneously affects cell-matrix interactions and growth factor activity in these cells. Increasing BMP-1 led to a loss of cell adhesion that depended on the matricellular glycoprotein thrombospondin-1 (TSP-1). BMP-1 cleaved TSP-1 between the VWFC/procollagen-like domain and the type 1 repeats that mediate several key TSP-1 functions. This cleavage induced the release of TSP-1 C-terminal domains from the extracellular matrix and abolished its previously described multisite cooperative interactions with heparan sulfate proteoglycans and CD36 on HT1080 cells. In addition, BMP-1-dependent proteolysis potentiated the TSP-1-mediated activation of latent transforming growth factor-β (TGF-β), leading to increased signaling through the canonical SMAD pathway. In primary human corneal stromal cells (keratocytes), endogenous BMP-1 cleaved TSP-1, and the addition of exogenous BMP-1 enhanced cleavage, but this had no substantial effect on cell adhesion. Instead, processed TSP-1 promoted the differentiation of keratocytes into myofibroblasts and stimulated production of the myofibroblast marker α-SMA, consistent with the presence of processed TSP-1 in human corneal scars. Our results indicate that BMP-1 can both trigger the disruption of cell adhesion and stimulate TGF-β signaling in TSP-1-rich microenvironments, which has important potential consequences for wound healing and tumor progression.
JUL 2020
DOI : 10.1126/scisignal.aba3880
Pubmed ID : 32636307
- CKD Increases Carbonylation of HDL and Is Associated with Impaired Antiaggregant Properties.
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Florens, N; Calzada, C; Lemoine, S; Boulet, MM; Guillot, N; Barba, C; Roux, J; Delolme, F; Page, A; Poux, JM; Laville, M; Moulin, P; Soulère, L; Guebre-Egziabher, F; Juillard, L; Soulage, CO
- J AM SOC NEPHROL
- Collaborative platform: PSF
Abstract :
HDL from CKD rabbits and patients on hemodialysis had HNE adducts. The percentage of platelet aggregation or activation induced by collagen was significantly higher when platelets were incubated with HDL from CKD rabbit and hemodialysis groups than with HDL from the control group. In both rabbits and humans, platelet aggregation and activation were significantly higher in the presence of HNE-modified HDL than with HDL from their respective controls. Incubation of platelets with a blocking antibody directed against CD36 or with a pharmacologic inhibitor of SRC kinases restored the antiaggregative phenotype in the presence of HDL from CKD rabbits, patients on hemodialysis and peritoneal dialysis, and HNE-modified HDL.
JUL 2020
DOI : 10.1681/ASN.2019111205
Pubmed ID : 32518085
- The Inflammasome Adaptor ASC Delays UV-Induced Skin Tumorigenesis in Beta HPV38 E6 and E7 Transgenic Mice.
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Magnotti, F; Valsesia, S; Gupta, P; Flechtenmacher, C; Contard, P; Viarisio, D; Venuti, A; Wencker, M; Tommasino, M; Marvel, J; Henry T
- J INVEST DERMATOL
- Collaborative platform: PBES
Abstract :
Non-melanoma skin cancer is the most common cancer in fair-skinned adults and comprises basal cell carcinoma and squamous cell carcinoma (SCC). Its incidence reaches 1 of 100 person-year in Australia and is still on the rise worldwide. The development of non-melanoma skin cancer depends on genetic and environmental factors. Particularly, there is clear evidence that cumulative sun exposure contributes to the development of non-melanoma skin cancer. In addition, the fact that immunosuppression strongly increases the susceptibility to non-melanoma skin cancer supports the role of infectious agents in this pathologic condition. Indeed, a subgroup of cutaneous human papillomavirus (HPV) types belonging to genus Beta of the HPV phylogenetic tree has been implicated as the etiological factor of cutaneous SCC.
MAY 2020
DOI : 10.1016/j.jid.2020.04.028
Pubmed ID : 32470340
- Taking Advantage of Oxidation to Characterize Thiol-Containing Polymer Chains by MALDI-TOF Mass Spectrometry.
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Gaillot, C; Delolme, F; Fabre, L; Charreyre, MT; Ladavière, C; Favier A
- ANAL CHEM
- Collaborative platform: PSF
Abstract :
MALDI-TOF mass spectrometry analyses revealed the oxidation of thiol-containing polymer chain-ends during sample preparation using THF as solvent. In these conditions, the extent of oxidation was hardly reproducible, and led to various types of oxidized compounds. Preparing the samples at the last minute using commercial THF stabilized with an antioxidant led to more reproducible results, with the least oxidation. However, it is demonstrated herein that thiol oxidation can be advantageously taken into profit to further ascertain the presence of the thiol at the polymer chain-end. To force thiol oxidation we used THF without any antioxidant stabilizer, thus more prone to form peroxides. Thiol-containing polymer chains can thereby be indirectly evidenced by the formation of oxidation products such as chain-chain disulfide bonds and sulfonic acid chains-ends. More importantly, in these oxidizing conditions and in the negative mode, sulfonic acid-terminated polymer chains can be more sensitively detected than thiol ones (the low pKa of sulfonic acids facilitating their anionization in MALDI source). In conclusion, performing MALDI-TOF mass spectrometry analyses in oxidizing conditions, as complement to regular analyses, was found to be very useful for the chain-end identification of different thiol-containing polymer chains.
MAR 2020
DOI : 10.1021/acs.analchem.9b05207
Pubmed ID : 31990181
- Gradient in cytoplasmic pressure in germline cells controls overlying epithelial cell morphogenesis
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Lamiré, LA; Milani, P; Runel, G; Kiss, A; Arias, L; Vergier, B; de Bossoreille, S; Das, P; Cluet, D; Boudaoud, A; Grammont,M
- PLOS BIOL
- Acknowledged platforms: PLATIM, Arthrotools
Abstract :
It is unknown how growth in one tissue impacts morphogenesis in a neighboring tissue. To address this, we used the Drosophila ovarian follicle, in which a cluster of 15 nurse cells and a posteriorly located oocyte are surrounded by a layer of epithelial cells. It is known that as the nurse cells grow, the overlying epithelial cells flatten in a wave that begins in the anterior. Here, we demonstrate that an anterior to posterior gradient of decreasing cytoplasmic pressure is present across the nurse cells and that this gradient acts through TGFβ to control both the triggering and the progression of the wave of epithelial cell flattening. Our data indicate that intrinsic nurse cell growth is important to control proper nurse cell pressure. Finally, we reveal that nurse cell pressure and subsequent TGFβ activity in the stretched cells combine to increase follicle elongation in the anterior, which is crucial for allowing nurse cell growth and pressure control. More generally, our results reveal that during development, inner cytoplasmic pressure in individual cells has an important role in shaping their neighbors.
NOV 2020
DOI : 10.1371/journal.pbio.3000940
Pubmed ID : 33253165
- Spen modulates lipid droplet content in adult Drosophila glial cells and protects against paraquat toxicity
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Girard, V; Goubard,V; Querenet, M; Seugnet, L; Pays,L; Nataf,S; Dufourd, E; Cluet,D; Mollereau, B; Davoust, N
- SCI REP
- Acknowledged platform: PLATIM
Abstract :
Glial cells are early sensors of neuronal injury and can store lipids in lipid droplets under oxidative stress conditions. Here, we investigated the functions of the RNA-binding protein, SPEN/SHARP, in the context of Parkinson’s disease (PD). Using a data-mining approach, we found that SPEN/SHARP is one of many astrocyte-expressed genes that are significantly differentially expressed in the substantia nigra of PD patients compared with control subjects. Interestingly, the differentially expressed genes are enriched in lipid metabolism-associated genes. In a Drosophila model of PD, we observed that flies carrying a loss-of-function allele of the ortholog split-ends (spen) or with glial cell-specific, but not neuronal-specific, spen knockdown were more sensitive to paraquat intoxication, indicating a protective role for Spen in glial cells. We also found that Spen is a positive regulator of Notch signaling in adult Drosophila glial cells. Moreover, Spen was required to limit abnormal accumulation of lipid droplets in glial cells in a manner independent of its regulation of Notch signaling. Taken together, our results demonstrate that Spen regulates lipid metabolism and storage in glial cells and contributes to glial cell-mediated neuroprotection.
NOV 2020
DOI : 10.1038/s41598-020-76891-9
Pubmed ID : 33208773
- The interplays between Crimean-Congo hemorrhagic fever virus (CCHFV) M segment-encoded accessory proteins and structural proteins promote virus assembly and infectivity
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Freitas, N; Enguehard, M; Denolly, S; Levy, C; Neveu, G; Lerolle, S; Devignot, S; Weber, F; Bergeron, C; Legros,V; Cosset, FL
- PLOS PATHOG
- Acknowledged platforms: PLATIM, Cytometry, AGC
Abstract :
Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne orthonairovirus that has become a serious threat to the public health. CCHFV has a single-stranded, tripartite RNA genome composed of L, M, and S segments. Cleavage of the M polyprotein precursor generates the two envelope glycoproteins (GPs) as well as three secreted nonstructural proteins GP38 and GP85 or GP160, representing GP38 only or GP38 linked to a mucin-like protein (MLD), and a double-membrane-spanning protein called NSm. Here, we examined the relevance of each M-segment non-structural proteins in virus assembly, egress and infectivity using a well-established CCHFV virus-like-particle system (tc-VLP). Deletion of MLD protein had no impact on infectivity although it reduced by 60% incorporation of GPs into particles. Additional deletion of GP38 abolished production of infectious tc-VLPs. The loss of infectivity was associated with impaired Gc maturation and exclusion from the Golgi, showing that Gn is not sufficient to target CCHFV GPs to the site of assembly. Consistent with this, efficient complementation was achieved in cells expressing MLD-GP38 in trans with increased levels of preGc to Gc conversion, co-targeting to the Golgi, resulting in particle incorporation and restored infectivity. Contrastingly, a MLD-GP38 variant retained in the ER allowed preGc cleavage but failed to rescue miss-localization or infectivity. NSm deletion, conversely, did not affect trafficking of Gc but interfered with Gc processing, particle formation and secretion. NSm expression affected N-glycosylation of different viral proteins most likely due to increased speed of trafficking through the secretory pathway. This highlights a potential role of NSm in overcoming Golgi retention and facilitating CCHFV egress. Thus, deletions of GP38 or NSm demonstrate their important role on CCHFV particle production and infectivity. GP85 is an essential viral factor for preGc cleavage, trafficking and Gc incorporation into particles, whereas NSm protein is involved in CCHFV assembly and virion secretion.
SEP 2020
DOI : 10.1371/journal.ppat.1008850
Pubmed ID : 32956404
- The Lysine Specific Demethylase-1 Negatively Regulates the COL9A1 Gene in Human Articular Chondrocytes.
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Durand, AL; Dufour, A; Aubert-Foucher, E; Oger-Desfeux, C; Pasdeloup, M; Lustig, S; Servien, E; Vaz,G; Perrier-Groult, E; Mallein-Gerin, F; Lafont, JE
- INT J MOL SCI
- Acknowledged platforms: AGC
Abstract :
Osteoarthritis (OA) is a degenerative disease of the joints which is associated with an impaired production of the cartilage matrix by the chondrocytes. Here, we investigated the role of Lysine-Specific Demethylase-1 (LSD1), a chromatin remodeling enzyme whose role in articular chondrocytes was previously associated with a catabolic activity and which is potentially involved during OA. Following a loss of function strategy and RNA sequencing analysis, we detail the genes which are targeted by LSD1 in human articular chondrocytes and identify COL9A1, a gene encoding the α1 chain of the cartilage-specific type IX collagen, as negatively regulated by LSD1. We show that LSD1 interacts with the transcription factor SOX9 and is recruited to the promoter of COL9A1. Interestingly, we observe that OA cartilage displays stronger LSD1 immunostaining compared with normal, and we demonstrate that the depletion of LSD1 in OA chondrocytes prevents the decrease in COL9A1 following Il-1β treatment. These results suggest LSD1 is a new regulator of the anabolic activity of articular chondrocytes potentially destabilizing the cartilage matrix, since it negatively regulates COL9A1, a gene encoding a crucial anchoring collagen molecule. This newly identified role played by LSD1 may thus participate in the alteration of the cartilage matrix during OA.
AUG 2020
DOI : 10.3390/ijms21176322
Pubmed ID : 32878268
- Cranial evolution in the extinct Rodrigues Island owl Otus murivorus (Strigidae), associated with unexpected ecological adaptations
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Duhamel,A; Hume,JP; Guenser, P; Salaviale, C; Louchart, A
- SCI REP
- Acknowledged platform: ImmOs
Abstract :
Island birds that were victims of anthropic extinctions were often more specialist species, having evolved their most distinctive features in isolation, making the study of fossil insular birds most interesting. Here we studied a fossil cranium of the ‘giant’ extinct scops owl Otus murivorus from Rodrigues Island (Mascarene Islands, southwestern Indian Ocean), to determine any potential unique characters. The fossil and extant strigids were imaged through X-ray microtomography, providing 3D views of external and internal (endocast, inner ear) cranial structures. Geometric morphometrics and analyses of traditional measurements yielded new information about the Rodrigues owl’s evolution and ecology. Otus murivorus exhibits a 2-tier “lag behind” phenomenon for cranium and brain evolution, both being proportionately small relative to increased body size. It also had a much more developed olfactory bulb than congeners, indicating an unexpectedly developed olfactory sense, suggesting a partial food scavenging habit. In addition, O. murivorus had the eyes placed more laterally than O. sunia, the species from which it was derived, probably a side effect of a small brain; rather terrestrial habits; probably relatively fearless behavior; and a less vertical posture (head less upright) than other owls (this in part an allometric effect of size increase). These evolutionary features, added to gigantism and wing reduction, make the extinct Rodrigues owl’s evolution remarkable, and with multiple causes..
AUG 2020
DOI : 10.1038/s41598-020-69868-1
Pubmed ID : 32820225
- Structure and catalytic regulation of Plasmodium falciparum IMP specific nucleotidase
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Carrique, L; Ballut, L; Shukla, A; Varma, N; Ravi, R; Violot, S; Srinivasan, B; Ganeshappa, UT; Kulkarni, S; Balaram, H; Aghajari, N
- NAT COMM
- Acknowledged platform: PSF
Abstract :
Plasmodium falciparum (Pf) relies solely on the salvage pathway for its purine nucleotide requirements, making this pathway indispensable to the parasite. Purine nucleotide levels are regulated by anabolic processes and by nucleotidases that hydrolyse these metabolites into nucleosides. Certain apicomplexan parasites, including Pf, have an IMP-specific-nucleotidase 1 (ISN1). Here we show, by comprehensive substrate screening, that PfISN1 catalyzes the dephosphorylation of inosine monophosphate (IMP) and is allosterically activated by ATP. Crystal structures of tetrameric PfISN1 reveal complex rearrangements of domain organization tightly associated with catalysis. Immunofluorescence microscopy and expression of GFP-fused protein indicate cytosolic localization of PfISN1 and expression in asexual and gametocyte stages of the parasite. With earlier evidence on isn1 upregulation in female gametocytes, the structures reported in this study may contribute to initiate the design for possible transmission-blocking agents.
JUN 2020
DOI :10.1038/s41467-020-17013-x
Pubmed ID : 32591529
- Metabolic Cooperation among Commensal Bacteria Supports Drosophila Juvenile Growth under Nutritional Stress
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Consuegra, J; Grenier, T; Akherraz, H; Rahioui, I; Gervais, H; da Silva, P; Leulier, F
- ISCIENCE
- Acknowledged platform: ArthroTools
Abstract :
The gut microbiota shapes animal growth trajectory in stressful nutritional environments, but the molecular mechanisms behind such physiological benefits remain poorly understood. The gut microbiota is mostly composed of bacteria, which construct metabolic networks among themselves and with the host. Until now, how the metabolic activities of the microbiota contribute to host juvenile growth remains unknown. Here, using Drosophila as a host model, we report that two of its major bacterial partners, Lactobacillus plantarum and Acetobacter pomorum, engage in a beneficial metabolic dialogue that boosts host juvenile growth despite nutritional stress. We pinpoint that lactate, produced by L. plantarum, is utilized by A. pomorum as an additional carbon source, and A. pomorum provides essential amino acids and vitamins to L. plantarum. Such bacterial cross-feeding provisions a set of anabolic metabolites to the host, which may foster host systemic growth despite poor nutrition.
JUN 2020
DOI :10.1016/j.isci.2020.101232
Pubmed ID : 31068585
- Presynaptic APP levels and synaptic homeostasis are regulated by Akt phosphorylation of huntingtin
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Bruyère, J; Abada, YS; Vitet, H; Fontaine, G; Deloulme, JC; Cès, A; Denarier, E; Pernet-Gallay, K; Andrieux, A; Humbert,S; Potier,MC; Delatour, B; Saudou, F
- ELIFE
- Acknowledge platform: Vectorology
Abstract :
Studies have suggested that amyloid precursor protein (APP) regulates synaptic homeostasis, but the evidence has not been consistent. In particular, signaling pathways controlling APP transport to the synapse in axons and dendrites remain to be identified. Having previously shown that Huntingtin (HTT), the scaffolding protein involved in Huntington’s disease, regulates neuritic transport of APP, we used a microfluidic corticocortical neuronal network-on-a-chip to examine APP transport and localization to the pre- and post-synaptic compartments. We found that HTT, upon phosphorylation by the Ser/Thr kinase Akt, regulates APP transport in axons but not dendrites. Expression of an unphosphorylatable HTT decreased axonal anterograde transport of APP, reduced presynaptic APP levels, and increased synaptic density. Ablating in vivo HTT phosphorylation in APPPS1 mice, which overexpress APP, reduced presynaptic APP levels, restored synapse number and improved learning and memory. The Akt-HTT pathway and axonal transport of APP thus regulate APP presynaptic levels and synapse homeostasis.
MAY 2020
DOI : 10.7554/eLife.56371
Pubmed ID : 32452382
- Insight into the Lifestyle of Amoeba Willaertia magna during Bioreactor Growth Using Transcriptomics and Proteomics
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Hasni, I; Decloquement, P; Demanèche, S; Mameri, RM; Abbe, O; Colson, P; La Scola, B
- MICROORGANISMS
- Acknowledged platform: PSF
Abstract :
To gain insight into the biology of NK cells, others and we previously identified the NK-cell signature, defined as the set of transcripts which expression is highly enriched in these cells compared to other immune subtypes. The transcript encoding the Serine/threonine/tyrosine kinase 1 (Styk1) is part of this signature. However, the role of Styk1 in the immune system is unknown. Here, we report the generation of a novel transgenic mouse model, in which Styk1 expression is invalidated and replaced by an EGFP reporter cassette. We demonstrated that Styk1 expression is a hallmark of NK cells and other NK1.1 expressing cells such as liver type 1 innate lymphoid cells (ILC1) and NK1.1(+) gamma delta T cells. Styk1 expression is maintained by IL-15 in NK cells and negatively correlates with the expression of educating NK-cell receptors. Analysis of phosphorylation levels of mTOR substrates suggested that Styk1 could moderately contribute to the activity of the PI3K/Akt/mTOR pathway. However, Styk1-deficient NK cells develop normally and have normal in vitro and in vivo effector functions. Thus Styk1 expression is a hallmark of NK cells, ILC1 and NK1.1(+) T cells but is dispensable for their development and immune functions.
MAY 2020
DOI : 10.3390/microorganisms8050771
Pubmed ID : 32455615
- Viral infection impacts transposable element transcript amounts in Drosophila
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Roy, M; Viginier, B; Saint-Michel, E; Arnaud, F; Ratinier, M; Fablet, M
- PROC NATL ACAD SCI U S A
- Acknowledged platform: ArthroTools
Abstract :
Transposable elements (TEs) are genomic parasites that are found in all genomes, some of which display sequence similarity to certain viruses. In insects, TEs are controlled by the Piwi-interacting small interfering RNA (piRNA) pathway in gonads, while the small interfering RNA (siRNA) pathway is dedicated to TE somatic control and defense against viruses. So far, these two small interfering RNA pathways are considered to involve distinct molecular effectors and are described as independent. Using Sindbis virus (SINV) in Drosophila, here we show that viral infections affect TE transcript amounts via modulations of the piRNA and siRNA repertoires, with the clearest effects in somatic tissues. These results suggest that viral acute or chronic infections may impact TE activity and, thus, the tempo of genetic diversification. In addition, these results deserve further evolutionary considerations regarding potential benefits to the host, the virus, or the TEs
JUN 2020
DOI : 10.1073/pnas.2006106117
Pubmed ID : 32434916
- Temporal integration of auxin information for the regulation of patterning
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Galvan-Ampudia, CS; Cerutti, G; Legrand, J; Brunoud, G; Martin-Arevalillo, R; Azais, R; Bayle, V; Moussu, S; Wenzl, C; Jaillais, Y; Lohmann, JU; Godin, C; Vernoux, T
- ELIFE
- Acknowledged platform: PLATIM
Abstract :
Positional information is essential for coordinating the development of multicellular organisms. In plants, positional information provided by the hormone auxin regulates rhythmic organ production at the shoot apex, but the spatio-temporal dynamics of auxin gradients is unknown. We used quantitative imaging to demonstrate that auxin carries high-definition graded information not only in space but also in time. We show that, during organogenesis, temporal patterns of auxin arise from rhythmic centrifugal waves of high auxin travelling through the tissue faster than growth. We further demonstrate that temporal integration of auxin concentration is required to trigger the auxin-dependent transcription associated with organogenesis. This provides a mechanism to temporally differentiate sites of organ initiation and exemplifies how spatio-temporal positional information can be used to create rhythmicity.
MAY 2020
DOI : 10.7554/eLife.55832
Pubmed ID : 32379043
- H2A.Z is dispensable for both basal and activated transcription in post-mitotic mouse muscles
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Belotti, E; Lacoste, N; Simonet, T; Papin, C; Padmanabhan,K; Scionti, I; Gangloff, YG; Ramos, L; Dalkara,D; Hamiche, A; Dimitrov, S; Schaeffer, L
- NUCLEIC ACIDS RES
- Acknowledged platform: PBES
Abstract :
While the histone variant H2A.Z is known to be required for mitosis, it is also enriched in nucleosomes surrounding the transcription start site of active promoters, implicating H2A.Z in transcription. However, evidence obtained so far mainly rely on correlational data generated in actively dividing cells. We have exploited a paradigm in which transcription is uncoupled from the cell cycle by developing an in vivo system to inactivate H2A.Z in terminally differentiated post-mitotic muscle cells. ChIP-seq, RNA-seq and ATAC-seq experiments performed on H2A.Z KO post-mitotic muscle cells show that this histone variant is neither required to maintain nor to activate transcription. Altogether, this study provides in vivo evidence that in the absence of mitosis H2A.Z is dispensable for transcription and that the enrichment of H2A.Z on active promoters is a marker but not an active driver of transcription.
MAY 2020
DOI : 10.1093/nar/gkaa157
Pubmed ID : 32266374
- Development of a Reverse Genetics System for Toscana Virus (Lineage A)
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Alexander, AJT; Confort, MP; Desloire, S; Dunlop, JI; Kuchi, S; Sreenu, VB; Mair, D; Wilkie, GS; Da Silva Filipe, A; Brennan, B; Ratinier, M; Arnaud, F; Kohl, A
- VIRUSES
- Acknowledged platform:
Abstract :
Toscana virus (TOSV) is a Phlebovirus in the Phenuiviridae family, order Bunyavirales, found in the countries surrounding the Mediterranean. TOSV is an important cause of seasonal acute meningitis and encephalitis within its range. Here, we determined the full sequence of the TOSV strain 1500590, a lineage A virus obtained from an infected patient (Marseille, 2007) and used this in combination with other sequence information to construct functional cDNA plasmids encoding the viral L, M, and S antigenomic sequences under the control of the T7 RNA promoter to recover recombinant viruses. Importantly, resequencing identified two single nucleotide changes to a TOSV reference genome, which, when corrected, restored functionality to the polymerase L and made it possible to recover infectious recombinant TOSV (rTOSV) from cDNA, as well as establish a minigenome system. Using reverse genetics, we produced an NSs-deletant rTOSV and also obtained viruses expressing reporter genes instead of NSs. The availability of such a system assists investigating questions that require genetic manipulation of the viral genome, such as investigations into replication and tropism, and beyond these fundamental aspects, also the development of novel vaccine design strategies.
APR 2020
DOI : 10.3390/v12040411
Pubmed ID : 32272808
- Drosophila-associated bacteria differentially shape the nutritional requirements of their host during juvenile growth
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Consuegra, J; Grenier, T; Baa-Puyoulet, P; Rahioui, I; Akherraz, H; Gervais, H; Parisot, N; da Silva, P; Charles, H; Calevro, F; Leulier, F
- PLOS BIOL
- Acknowledged platform: ArthroTools
Abstract :
The interplay between nutrition and the microbial communities colonizing the gastrointestinal tract (i.e., gut microbiota) determines juvenile growth trajectory. Nutritional deficiencies trigger developmental delays, and an immature gut microbiota is a hallmark of pathologies related to childhood undernutrition. However, how host-associated bacteria modulate the impact of nutrition on juvenile growth remains elusive. Here, using gnotobiotic Drosophila melanogaster larvae independently associated with Acetobacter pomorumWJL (ApWJL) and Lactobacillus plantarumNC8 (LpNC8), 2 model Drosophila-associated bacteria, we performed a large-scale, systematic nutritional screen based on larval growth in 40 different and precisely controlled nutritional environments. We combined these results with genome-based metabolic network reconstruction to define the biosynthetic capacities of Drosophila germ-free (GF) larvae and its 2 bacterial partners. We first established that ApWJL and LpNC8 differentially fulfill the nutritional requirements of the ex-GF larvae and parsed such difference down to individual amino acids, vitamins, other micronutrients, and trace metals. We found that Drosophila-associated bacteria not only fortify the host’s diet with essential nutrients but, in specific instances, functionally compensate for host auxotrophies by either providing a metabolic intermediate or nutrient derivative to the host or by uptaking, concentrating, and delivering contaminant traces of micronutrients. Our systematic work reveals that beyond the molecular dialogue engaged between the host and its bacterial partners, Drosophila and its associated bacteria establish an integrated nutritional network relying on nutrient provision and utilization.
MAR 2020
DOI : 10.1371/journal.pbio.3000681
Pubmed ID : 32196485
- Generation of an Fsp1 (fibroblast‐specific protein 1)‐Flpo transgenic mouse strain
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Cardot‐Ruffino, V; Chauvet, V; Caligaris, C; Bertrand‐Chapel, A; Chuvin, N; Pommier, RM; Valcourt, U; Vincent, D; Martel, S; Aires, S; Kaniewski, B; Dubus, P; Cassier, P; Sentis, S; Bartholin, L
- GENESIS 2020
- Acknowledged platform: PBES
Abstract :
Recombination systems represent a major breakthrough in the field of genetic model engineering. The Flp recombinases (Flp, Flpe, and Flpo) bind and cleave DNA Frt sites. We created a transgenic mouse strain ([Fsp1‐Flpo]) expressing the Flpo recombinase in fibroblasts. This strain was obtained by random insertion inside mouse zygotes after pronuclear injection. Flpo expression was placed under the control of the promoter of Fsp1 (fibroblast‐specific protein 1) gene, whose expression starts after gastrulation at Day 8.5 in cells of mesenchymal origin. We verified the correct expression and function of the Flpo enzyme by several ex vivo and in vivo approaches. The [Fsp1‐Flpo] strain represents a genuine tool to further target the recombination of transgenes with Frt sites specifically in cells of mesenchymal origin or with a fibroblastic phenotype.
MAY 2020
DOI : 10.1002/dvg.23359
Pubmed ID : 32191380
- Noninvasive monitoring of liver metastasis development via combined multispectral photoacoustic imaging and fluorescence diffuse optical tomography
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Lavaud, J; Henry, M; Gayet, P; Fertin, A; Vollaire, J; Usson, Y; Coll, JL; Josserand, V
- INT J BIOL SCI
- Acknowledged platform: Vectorology
Abstract :
Contrast-agent-free PAI reflected tumor angiogenesis and gradual hypoxia during metastasis development. Multispectral PAI enabled noninvasive real-time monitoring of ICG blood pharmacokinetics, which demonstrated tumor-related liver dysfunction. Both PAI and fluorescence ICG signals were clearly modified in metastasis-bearing livers but did not allow for differentiation between different disease stages. In contrast, there was a significant improvement achieved by using the tumor-specific marker Angiostamp800, which provided gradually increasing PAI and fDOT signals during metastasis development.
MAR 2020
DOI : 10.7150/ijbs.40896
Pubmed ID : 32226306
- TTC12 Loss-of-Function Mutations Cause Primary Ciliary Dyskinesia and Unveil Distinct Dynein Assembly Mechanisms in Motile Cilia Versus Flagella
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Thomas, L; Bouhouche, K; Whitfield, M; Thouvenin, G; Coste, A; Louis, B; Szymanski, L; Bequignon, E; Papon, JF; Castelli, M; Lemullois, M; Dhalluin, X; Drouin-Garraud, V; Montantin, G; Tissier, S; Duquesnoy, P; Copin, B; Dastot, F; Couvet, S; Barbotin, AL; Faucon, C; Honore, I; Maitre, B; Beydon, N; Tamalet, A; Rives, N; Koll, F; Escudier, E; Tassin, AM; Touré, A; Mitchell,V; Amselem, S; Legendre, M
- AM J HUM GENET
- Acknowledged platform: Vectorology
Abstract :
Cilia and flagella are evolutionarily conserved organelles whose motility relies on the outer and inner dynein arm complexes (ODAs and IDAs). Defects in ODAs and IDAs result in primary ciliary dyskinesia (PCD), a disease characterized by recurrent airway infections and male infertility. PCD mutations in assembly factors have been shown to cause a combined ODA-IDA defect, affecting both cilia and flagella. We identified four loss-of-function mutations in TTC12, which encodes a cytoplasmic protein, in four independent families in which affected individuals displayed a peculiar PCD phenotype characterized by the absence of ODAs and IDAs in sperm flagella, contrasting with the absence of only IDAs in respiratory cilia. Analyses of both primary cells from individuals carrying TTC12 mutations and human differentiated airway cells invalidated for TTC12 by a CRISPR-Cas9 approach revealed an IDA defect restricted to a subset of single-headed IDAs that are different in flagella and cilia, whereas TTC12 depletion in the ciliate Paramecium tetraurelia recapitulated the sperm phenotype. Overall, our study, which identifies TTC12 as a gene involved in PCD, unveils distinct dynein assembly mechanisms in human motile cilia versus flagella.
FEB 2020
DOI : 10.1016/j.ajhg.2019.12.010
Pubmed ID : 31978331
- Huntingtin phosphorylation governs BDNF homeostasis and improves the phenotype of Mecp2 knockout mice
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Ehinger, Y; Bruyère, J; Panayotis, N; Abada, YS; Borloz, E; Matagne, V; Scaramuzzino, C; Vitet, H; Delatour, B; Saidi, L; Villard, L; Saudou, F; Roux, JC;
- EMBO MOL MED
- Acknowledged platform: Vectorology
Abstract :
Mutations in the X‐linked MECP2 gene are responsible for Rett syndrome (RTT), a severe neurological disorder for which there is no treatment. Several studies have linked the loss of MeCP2 function to alterations of brain‐derived neurotrophic factor (BDNF) levels, but non‐specific overexpression of BDNF only partially improves the phenotype of Mecp2‐deficient mice. We and others have previously shown that huntingtin (HTT) scaffolds molecular motor complexes, transports BDNF‐containing vesicles, and is under‐expressed in Mecp2 knockout brains. Here, we demonstrate that promoting HTT phosphorylation at Ser421, either by a phospho‐mimetic mutation or inhibition of the phosphatase calcineurin, restores endogenous BDNF axonal transport in vitro in the corticostriatal pathway, increases striatal BDNF availability and synaptic connectivity in vivo, and improves the phenotype and the survival of Mecp2 knockout mice—even though treatments were initiated only after the mice had already developed symptoms. Stimulation of endogenous cellular pathways may thus be a promising approach for the treatment of RTT patients.
FEB 2020
DOI : 10.15252/emmm.201910889
Pubmed ID : 31913581