Crowded chromatin is not sufficient for heterochromatin formation and not required for its maintenance

International audienceIn contrast to cytoplasmic organelles, which are usually separated from the rest of the cell by phospholipid membranes, nuclear compartments are readily accessible to diffusing proteins and must rely on different mechanisms to maintain their integrity. Specific interactions between scaffolding proteins are known to have important roles for the formation and maintenance of nuclear structures. General physical mechanisms such as molecular crowding, phase separation or colloidal behavior have also been suggested, but their physiological significance remains uncertain. For macromolecular crowding, a role in the maintenance of nucleoli and promyelocytic leukemia (PML) nuclear bodies has been shown. Here, we tested whether a modulation of the compaction state of chromatin, which directly influences the local crowding state, has an impact on the formation and maintenance of densely packed heterochromatin. By osmotic perturbations, we could modify the packing state of chromatin in a controlled manner and show that chromatin compaction, which is associated with increased crowding conditions, is not, per se, sufficient to initiate the formation of new bona fide heterochromatin structures nor is it necessary to maintain already established heterochromatin domains. In consequence, if an increase in crowding induced by chromatin compaction maybe an early step in heterochromatin formation, specific protein-protein interactions are nevertheless required to make heterochromatin long lasting and independent of the crowding state

Hardware communication refinement in digital signal processing, modelling issues

In this paper we present the different modelling problems which a Digital Signal Processing (DSP) application designer has to tackle while refining an abstract specification relying on coarse grain data (e.g. matrices) toward a hardware implementation model relying on fine grain data (e.g. scalar). To address this problematic, we propose a modelling framework which can be used to refine an algorithm specified with coarse grain interfaces to a form which allow, from the functionnality point of view, to model all its fine grain hardware implmentation

Seed Spillage from Grain Trailers on Road Verges during Oilseed Rape Harvest: An Experimental Survey

Context: Anthropogenic vectors enhance the natural dispersal capacity of plant seeds significantly in terms of quantity and distance. Human-mediated seed dispersal (i.e. anthropochory) greatly increases the dispersal of crop species across agroecosystems. In the case of oilseed rape (OSR), spillage of seeds from grain trailers during harvest has never been quantified. Methods: Our experimental approach involved establishing 85 seed trap-sites on the road verges of an agricultural area around the grain silo of Selommes (Loir-et-Cher, France). We recorded OSR spillage during harvest and applied a linear model to the data. Results: The amount of seed spilled was related positively to the area of the OSR fields served by the road, whereas the amount of seed spilled decreased with other variables, such as distance from the trap-site to the verge of the road and to the nearest field. The distance to the grain silo, through local and regional effects, affected seed loss. Local effects from fields adjacent to the road resulted in a cumulative spillage on one-lane roads. On two-lane roads, spillage was nearly constant whatever the distance to the silo due to a mixture of these local effects and of grain trailers that joined the road from more distant fields. From the data, we predicted the number of seeds lost from grain trailers on one road verge in the study area. We predicted a total spillage of 2.05610 6 seeds (64.76610 5) along the road length, which represented

Photodermatoses médicamenteuses ( revue de la littérature et analyse des cas notifiés dans la Base Nationale de Pharmacovigilance du 1er Janvier 2010 au 30 Juin 2012. Place du pharmacien d officine)

Une photosensibilisation résulte de la rencontre au niveau cutané d une substance photosensibilisante et d une longueur d onde efficace. Elle se traduit par l apparition de lésions cutanées plus ou moins étendues et d aspect variable. Ce type d effet indésirable est déjà connu et est notamment observé avec les quinolones, le kétoprofène et l amiodarone. 417 observations de photosensibilisation médicamenteuses ont été recueillies et analysées à partir des données de la Base Nationale de Pharmacovigilance. Il en résulte que le pouvoir photosensibilisant des fibrates, de l acide tiaprofénique, des antidépresseurs ainsi que des diurétiques semble sous-estimé. L étude met aussi en avant la notion de réaction croisée qui peut survenir entre différentes classes médicamenteuses, parfois même avec certains cosmétiques. L octocrylène fait d ailleurs l objet de nombreuses discussions. Le diagnostic d une réaction de photosensibilisation repose avant tout sur la réalisation de photopatch-test. Malheureusement, comme le montre cette étude, cet examen de référence reste peu pratiqué. Le rôle du pharmacien d officine repose avant tout sur la prévention primaire du risque. Il assure un conseil officinal associé à chaque délivrance de produit au potentiel photosensibilisant en rappelant toutes les mesures de photoprotection aux patients.Photosensitivity follows from the meeting on the skin of a photosensitizer product and an effective wavelength. It results in the appearance of more or less widespread and variable aspect lesions on the skin. This type of side effect is known and is particularly observed with quinolones, ketoprofen and amiodaron. 417 observations of drug photosensitivity were collected and analyzed data from the National Pharmacovigilance Database. It follows that the photosensitizer potential to fibrates, tiaprofenic acid, antidepressants and diuretics seems underestimated. The study also highlights the notion of cross-reaction that can occur between different drug classes, sometimes even with some cosmetics. Octocrylene is also the subject of many discussions. Achieving photopatch testing remains the most effective way to diagnose photosensitivity. Unfortunately, this method is rarely used. The role of the pharmacist is primarily based on the primary prevention of risk. It provides advice associated with each product delivery to photosensitizing potential recalling all measures of photoprotection to patients.ANGERS-BU Médecine-Pharmacie (490072105) / SudocSudocFranceF

Rab27A and its effector MyRIP link secretory granules to F-actin and control their motion towards release sites

The GTPase Rab27A interacts with myosin-VIIa and myosin-Va via MyRIP or melanophilin and mediates melanosome binding to actin. Here we show that Rab27A and MyRIP are associated with secretory granules (SGs) in adrenal chromaffin cells and PC12 cells. Overexpression of Rab27A, GTPase-deficient Rab27A-Q78L, or MyRIP reduced secretory responses of PC12 cells. Amperometric recordings of single adrenal chromaffin cells revealed that Rab27A-Q78L and MyRIP reduced the sustained component of release. Moreover, these effects on secretion were partly suppressed by the actin-depolymerizing drug latrunculin but strengthened by jasplakinolide, which stabilizes the actin cortex. Finally, MyRIP and Rab27A-Q78L restricted the motion of SGs in the subplasmalemmal region of PC12 cells, as measured by evanescent-wave fluorescence microscopy. In contrast, the Rab27A-binding domain of MyRIP and a MyRIP construct that interacts with myosin-Va but not with actin increased the mobility of SGs. We propose that Rab27A and MyRIP link SGs to F-actin and control their motion toward release sites through the actin cortex

Ensconsin/Map7 promotes microtubule growth and centrosome separation in Drosophila neural stem cells.

International audienceThe mitotic spindle is crucial to achieve segregation of sister chromatids. To identify new mitotic spindle assembly regulators, we isolated 855 microtubule-associated proteins (MAPs) from Drosophila melanogaster mitotic or interphasic embryos. Using RNAi, we screened 96 poorly characterized genes in the Drosophila central nervous system to establish their possible role during spindle assembly. We found that Ensconsin/MAP7 mutant neuroblasts display shorter metaphase spindles, a defect caused by a reduced microtubule polymerization rate and enhanced by centrosome ablation. In agreement with a direct effect in regulating spindle length, Ensconsin overexpression triggered an increase in spindle length in S2 cells, whereas purified Ensconsin stimulated microtubule polymerization in vitro. Interestingly, ensc-null mutant flies also display defective centrosome separation and positioning during interphase, a phenotype also detected in kinesin-1 mutants. Collectively, our results suggest that Ensconsin cooperates with its binding partner Kinesin-1 during interphase to trigger centrosome separation. In addition, Ensconsin promotes microtubule polymerization during mitosis to control spindle length independent of Kinesin-1

EZH2 alterations in follicular lymphoma: biological and clinical correlations

International audienceThe histone methyltransferase EZH2 has an essential role in the development of follicular lymphoma (FL). Recurrent gain-of-function mutations in EZH2 have been described in 25% of FL patients and induce aberrant methylation of histone H3 lysine 27 (H3K27). We evaluated the role of EZH2 genomic gains in FL biology. Using RNA sequencing, Sanger sequencing and SNP-arrays, the mutation status, copy-number and gene-expression profiles of EZH2 were assessed in a cohort of 159 FL patients from the PRIMA trial. Immunohistochemical (IHC) EZH2 expression (n = 55) and H3K27 methylation (n = 63) profiles were also evaluated. In total, 37% of patients (59/159) harbored an alteration in the EZH2 gene (mutation n = 46, gain n = 23). Both types of alterations were associated with highly similar transcriptional changes, with increased proliferation programs. An H3K27me3/me2 IHC score fully distinguished mutated from wild-type samples, showing its applicability as surrogate for EZH2 mutation analysis. However, this score did not predict the presence of gains at the EZH2 locus. The presence of an EZH2 genetic alteration was an independent factor associated with a longer progression-free survival (hazard ratio 0.58, 95% confidence interval 0.36–0.93, P = 0.025). We propose that the copy-number status of EZH2 should also be considered when evaluating patient stratification and selecting patients for EZH2 inhibitor-targeted therapies

Genome-Scale Analysis of Mycoplasma agalactiae Loci Involved in Interaction with Host Cells

Mycoplasma agalactiae is an important pathogen of small ruminants, in which it causes contagious agalactia. It belongs to a large group of “minimal bacteria” with a small genome and reduced metabolic capacities that are dependent on their host for nutrients. Mycoplasma survival thus relies on intimate contact with host cells, but little is known about the factors involved in these interactions or in the more general infectious process. To address this issue, an assay based on goat epithelial and fibroblastic cells was used to screen a M. agalactiae knockout mutant library. Mutants with reduced growth capacities in cell culture were selected and 62 genomic loci were identified as contributing to this phenotype. As expected for minimal bacteria, “transport and metabolism” was the functional category most commonly implicated in this phenotype, but 50% of the selected mutants were disrupted in coding sequences (CDSs) with unknown functions, with surface lipoproteins being most commonly represented in this category. Since mycoplasmas lack a cell wall, lipoproteins are likely to be important in interactions with the host. A few intergenic regions were also identified that may act as regulatory sequences under co-culture conditions. Interestingly, some mutants mapped to gene clusters that are highly conserved across mycoplasma species but located in different positions. One of these clusters was found in a transcriptionally active region of the M. agalactiae chromosome, downstream of a cryptic promoter. A possible scenario for the evolution of these loci is discussed. Finally, several CDSs identified here are conserved in other important pathogenic mycoplasmas, and some were involved in horizontal gene transfer with phylogenetically distant species. These results provide a basis for further deciphering functions mediating mycoplasma-host interactions

Analyse des mouvements des granules de sécrétion à proximité de la membrane plasmique par microscopie de fluorescence à excitation par onde évanescente

Regulated hormone secretion is a multi-step process. Secretory granules (SG), which contain hormones, are formed by budding from the trans-golgi network and then migrate towards the cell periphery. Hormones are released by exocytosis when cells are stimulated. We applied total internal reflection fluorescence microscopy to study the dynamics of SG located in the subplasmalemmal region. We developed a motion analysis method to highlight transient behaviours along three-dimensional SG trajectories. The application of this method to the study of the effects of cytoskeletal drugs on SG dynamics and double labelling observations allowed us to associate each type of movement with a particular environment (SG linked to the plasma membrane, the actin filaments or the microtubules). We also studied the role of the complex formed by the proteins Rab27A, MyRIP and Myosin Va during the capture of SG in the cell periphery and their attachment to actin filaments.La sécrétion régulée d'hormones est un processus décomposable en plusieurs étapes. Les granules de sécrétion (GS) contenant ces hormones sont formés au niveau du réseau trans-golgien puis migrent jusqu'à la périphérie de la cellule. Ces hormones sont libérées dans le milieu extérieur par exocytose en cas de stimulation de la cellule. Grâce à l'observation des GS situés dans la région juxta-membranaire par microscopie de fluorescence à excitation par onde évanescente, les mouvements de ces organites ont été étudiés à l'échelle du granule unique. Une méthode d'analyse permettant la mise en évidence de comportements transitoires au sein des trajectoires tridimensionnelles des GS a été mise au point. Grâce à son application à l'étude des effets de drogues du cytosquelette et à des observations en double marquage, nous avons pu associer chaque type de mouvements décrit par les GS à un environnement particulier (GS lié à la membrane plasmique, au cytosquelette d'actine ou de microtubules). Nous avons de plus étudié le rôle du complexe protéique Rab27A/MyRIP/Myosine Va lors de la capture des GS en périphérie de la cellule et leur accrochage aux filaments d'actine

New Methodologies to Study DNA Repair Processes in Space and Time Within Living Cells

International audienceDNA repair requires a coordinated effort from an array of factors that play different roles in the DNA damage response from recognizing and signaling the presence of a break, creating a repair competent environment, and physically repairing the lesion. Due to the rapid nature of many of these events, live-cell microscopy has become an invaluable method to study this process. In this review we outline commonly used tools to induce DNA damage under the microscope and discuss spatio-temporal analysis tools that can bring added information regarding protein dynamics at sites of damage. In particular, we show how to go beyond the classical analysis of protein recruitment curves to be able to assess the dynamic association of the repair factors with the DNA lesions as well as the target-search strategies used to efficiently find these lesions. Finally, we discuss how the use of mathematical models, combined with experimental evidence, can be used to better interpret the complex dynamics of repair proteins at DNA lesions