Caspase-generated fragment of the Met receptor favors apoptosis via the intrinsic pathway independently of its tyrosine kinase activity

The receptor tyrosine kinase Met and its ligand, the hepatocyte growth factor, are essential to embryonic development, whereas the deregulation of Met signaling is associated with tumorigenesis. While ligand-activated Met promotes survival, caspase-dependent generation of the p40 Met fragment leads to apoptosis induction – hallmark of the dependence receptor. Although the survival signaling pathways induced by Met are well described, the pro-apoptotic signaling pathways are unknown. We show that, although p40 Met contains the entire kinase domain, it accelerates apoptosis independently of kinase activity. In cell cultures undergoing apoptosis, the fragment shows a mitochondrial localization, required for p40 Met-induced cell death. Fulminant hepatic failure induced in mice leads to the generation of p40 Met localized also in the mitochondria, demonstrating caspase cleavage of Met in vivo. According to its localization, the fragment induces mitochondrial permeabilization, which is inhibited by Bak silencing and Bcl-xL overexpression. Moreover, Met silencing delays mitochondrial permeabilization induced by an apoptotic treatment. Thus, the Met-dependence receptor in addition to its well-known role in survival signaling mediated by its kinase activity, also participates in the intrinsic apoptosis pathway through the generation of p40 Met – a caspase-dependent fragment of Met implicated in the mitochondrial permeabilization process

OA031-02. Regulatory T cells inhibit CD8 T cell proliferation in HIV-1 infection through CD39/adenosine pathway

International audiencen.

Communication Between the Cell Membrane and the Nucleus: Role of Protein Compartmentalization

Understanding how the information is conveyed from outside to inside the cell is a critical challenge for all biologists involved in signal transduction. The flow of information initiated by cell-cell and cell-extracellular matrix contacts is mediated by the formation of adhesion complexes involving multiple proteins. Inside adhesion complexes, connective membrane skeleton (CMS) proteins are signal transducers that bind to adhesion molecules, organize the cytoskeleton, and initiate biochemical cascades. Adhesion complex-mediated signal transduction ultimately directs the formation of supramolecular structures in the cell nucleus, as illustrated by the establishment of multi complexes of DNA-bound transcription factors, and the redistribution of nuclear structural proteins to form nuclear subdomains. Recently, several CMS proteins have been observed to travel to the cell nucleus, suggesting a distinctive role for these proteins in signal transduction. This review focuses on the nuclear translocation of structural signal transducers of the membrane skeleton and also extends our analysis to possible translocation of resident nuclear proteins to the membrane skeleton. This leads us to envision the communication between spatially distant cellular compartments (i.e., membrane skeleton and cell nucleus) as a bidirectional flow of information (a dynamic reciprocity) based on subtle multilevel structural and biochemical equilibria. At one level, it is mediated by the interaction between structural signal transducers and their binding partners, at another level it may be mediated by the balance and integration of signal transducers in different cellular compartments

New synchronization method for <i>Plasmodium falciparum</i>

&lt;b&gt;Background&lt;/b&gt;: Plasmodium falciparum is usually asynchronous during in vitro culture. Although various synchronization methods are available, they are not able to narrow the range of ages of parasites. A newly developed method is described that allows synchronization of parasites to produce cultures with an age range as low as 30 minutes. &lt;b&gt;Methods&lt;/b&gt;: Trophozoites and schizonts are enriched using Plasmion. The enriched late stage parasites are immobilized as a monolayer onto plastic Petri dishes using concanavalin A. Uninfected erythrocytes are placed onto the monolayer for a limited time period, during which time schizonts on the monolayer rupture and the released merozoites invade the fresh erythrocytes. The overlay is then taken off into a culture flask, resulting in a highly synchronized population of parasites. &lt;b&gt;Results&lt;/b&gt;: Plasmion treatment results in a 10- to 13-fold enrichment of late stage parasites. The monolayer method results in highly synchronized cultures of parasites where invasion has occurred within a very limited time window, which can be as low as 30 minutes. The method is simple, requiring no specialized equipment and relatively cheap reagents. &lt;b&gt;Conclusions&lt;/b&gt;: The new method for parasite synchronization results in highly synchronized populations of parasites, which will be useful for studies of the parasite asexual cell cycle

Shearmeter floats in the area of the WHOI Brazil Basin Tracer Release Experiment : technical and oceanographic data

Six drifting floats designed to measure shear were deployed in the vicinity of the Brazil Basin Tracer Release Experiment. The one-year long time series of oceanographic conditions obtained by the floats are for direct comparison with long-term tracer dispersion. The purpose of the tracer dispersion experiment was to study mixing of Antarctic Bottom Water at approximately 4000 m depth with less dense water above. Two of the floats returned shear records, one from about 1660 m depth and one from about 2800 m depth. Mean shear at 1660 m was 2.2 x 10 -3 s-1 with N = 1.1 cph, about 1.9 times the Garrett-Munk model amount. Mean shear at 2800 m was 1.1 x 10-3 with N = 0.5 cph, about 2.2 times Garrett-Munk. There was no apparent depth structure to the shear recorded by the near-bottom float moving over the mountainous seafloor. The two shear time series and the local tidal velocities were not strongly correlated, but the tide and shear series did have some similarities. Some variability in the 1660-m shear may be due to atmospheric forcing. Three floats deeper than 2800 m returned one-year long trajectories. Two trajectories were persistently eastward.Funding was provided by the National Science Foundation under Grant Nos. OCE-9416014 and OCE-9906685

Nanoengineered Astronomical Optics

We describe a technology for the fabrication of inexpensive and versatile mirrors through the use of a new type of nanoengineered optical material composed by the spreading of a self-assembling reflective colloidal film spread at the surface of a liquid. These new reflecting liquids offer interesting possibilities for astronomical instrumentation. For example, they can replace mercury in conventional rotating liquid mirrors. The main advantages offered include extremely low cost and, by coating a viscous liquid, the possibility of tilting the mirror by a few tens of degrees. We also have coated ferromagnetic liquids with these reflecting films. The resulting surfaces can be shaped by the application of a magnetic field, yielding reflecting surfaces that can have complicated shapes that can rapidly shift with time. These inexpensive and versatile optical elements could have numerous scientific and technological applications. Among possible astronomical applications, they could be used to make large inexpensive adaptive mirrors exhibiting strokes ranging from nanometers to several millimeters.Comment: Submitted to Astrophysical Journal Letters. 18 pages, 4 figure

Evaluation of electrothermal vaporization for sample introduction aiming at Cu isotopic analysis via multicollector-inductively coupled plasma mass spectrometry

A new method for Cu isotopic analysis was developed using a commercially available electrothermal vaporization (ETV) device coupled to multicollector-inductively coupled plasma mass spectrometry (MC-ICP-MS). The method demonstrated potential for the isotopic analysis of microsamples (e.g., 5 mu L) in a biological context. For example, Cu isotopic analysis of NIST 3114 (diluted to 1 mg L-1 Cu) using self-bracketing provided average delta Cu-65 values of 0.00 +/- 0.17%0 (2SD, n = 10) and internal precision values of 712 ppm. In order to achieve this level of accuracy and precision, it is critical to properly deal with the short transient signals generated by the ETV-MC-ICP-MS, which implies using point by point calculations and time lag detector correction (TDC), as well as a criterion to reject potential outliers. The results of this technique were compared with the results obtained via femtosecond-laser ablation-MC-ICPMS using the same pre-treated serum samples. No significant differences were observed among the results obtained in both cases, while external precision was 0.26%0 for ETV-MC-ICP-MS and 0.24%0 for fs-LA-MC-ICP-MS, expressed as median value of 2SD (n = 27), further proving the usefulness of the approach proposed in this context, as the use of ETV results in a more straightforward approach

Target identification of Mycobacterium tuberculosis phenotypic\textit{Mycobacterium tuberculosis phenotypic} hits using a concerted chemogenomic, biophysical and structural approach

Mycobacterium phenotypic hits are a good reservoir for new chemotypes for the treatment of tuberculosis. However, the absence of defined molecular targets and modes of action could lead to failure in drug development. Therefore, a combination of ligand-based and structure-based chemogenomic approaches followed by biophysical and biochemical validation have been used to identify targets for Mycobacterium tuberculosis phenotypic hits. Our approach identified EthR and InhA as targets for several hits, with some showing dual activity against these proteins. From the 35 predicted EthR inhibitors, eight exhibited an IC50 below 50 μM against M. tuberculosis EthR and three were confirmed to be also simultaneously active against InhA. Further hit validation was performed using X-ray crystallography yielding eight new crystal structures of EthR inhibitors. Although the EthR inhibitors attain their activity against M. tuberculosis by hitting yet undefined targets, these results provide new lead compounds that could be further developed to be used to potentiate the effect of EthA activated pro-drugs, such as ethionamide, thus enhancing their bactericidal effect.GM is grateful to the European Molecular Biology Laboratory and Marie Sklodowska-Curie Actions for funding this work. VM and MB acknowledge Bill & Melinda Gates Foundation [subcontract by the Foundation for the National Institutes of Health (NIH)] (OPP1024021). VM and MS acknowledge the European Community’s Seventh Framework Programme [grant number 260872]. GP would like to acknowledge the Wellcome Trust and the European Molecular Biology Laboratory for funding. JPO was funded by the member nation states of the European Molecular Biology Laboratory. TLB acknowledges The Wellcome Trust for funding and support (grant number 200814/Z/16/Z)