Tripartite phase separation of two signal effectors with vesicles priming B cell responsiveness.

Antibody-mediated immune responses rely on antigen recognition by the B cell antigen receptor (BCR) and the proper engagement of its intracellular signal effector proteins. Src homology (SH) 2 domain-containing leukocyte protein of 65 kDa (SLP65) is the key scaffold protein mediating BCR signaling. In resting B cells, SLP65 colocalizes with Cbl-interacting protein of 85 kDa (CIN85) in cytoplasmic granules whose formation is not fully understood. Here we show that effective B cell activation requires tripartite phase separation of SLP65, CIN85, and lipid vesicles into droplets via vesicle binding of SLP65 and promiscuous interactions between nine SH3 domains of the trimeric CIN85 and the proline-rich motifs (PRMs) of SLP65. Vesicles are clustered and the dynamical structure of SLP65 persists in the droplet phase in vitro. Our results demonstrate that phase separation driven by concerted transient interactions between scaffold proteins and vesicles is a cellular mechanism to concentrate and organize signal transducers

Concepts, Developments and Advanced Applications of the PAX Toolkit

The Physics Analysis eXpert (PAX) is an open source toolkit for high energy physics analysis. The C++ class collection provided by PAX is deployed in a number of analyses with complex event topologies at Tevatron and LHC. In this article, we summarize basic concepts and class structure of the PAX kernel. We report about the most recent developments of the kernel and introduce two new PAX accessories. The PaxFactory, that provides a class collection to facilitate event hypothesis evolution, and VisualPax, a Graphical User Interface for PAX objects

Radiation hardness of CMS pixel barrel modules

Pixel detectors are used in the innermost part of the multi purpose experiments at LHC and are therefore exposed to the highest fluences of ionising radiation, which in this part of the detectors consists mainly of charged pions. The radiation hardness of all detector components has thoroughly been tested up to the fluences expected at the LHC. In case of an LHC upgrade, the fluence will be much higher and it is not yet clear how long the present pixel modules will stay operative in such a harsh environment. The aim of this study was to establish such a limit as a benchmark for other possible detector concepts considered for the upgrade. As the sensors and the readout chip are the parts most sensitive to radiation damage, samples consisting of a small pixel sensor bump-bonded to a CMS-readout chip (PSI46V2.1) have been irradiated with positive 200 MeV pions at PSI up to 6E14 Neq and with 21 GeV protons at CERN up to 5E15 Neq. After irradiation the response of the system to beta particles from a Sr-90 source was measured to characterise the charge collection efficiency of the sensor. Radiation induced changes in the readout chip were also measured. The results show that the present pixel modules can be expected to be still operational after a fluence of 2.8E15 Neq. Samples irradiated up to 5E15 Neq still see the beta particles. However, further tests are needed to confirm whether a stable operation with high particle detection efficiency is possible after such a high fluence.Comment: Contribution to the 11th European Symposium on Semiconductor Detectors June 7-11, 2009 Wildbad Kreuth, German

Support varieties for selfinjective algebras

Support varieties for any finite dimensional algebra over a field were introduced by Snashall-Solberg using graded subalgebras of the Hochschild cohomology. We mainly study these varieties for selfinjective algebras under appropriate finite generation hypotheses. Then many of the standard results from the theory of support varieties for finite groups generalize to this situation. In particular, the complexity of the module equals the dimension of its corresponding variety, all closed homogeneous varieties occur as the variety of some module, the variety of an indecomposable module is connected, periodic modules are lines and for symmetric algebras a generalization of Webb's theorem is true

Requirement for beta-catenin in anterior-posterior axis formation in mice

The anterior-posterior axis of the mouse embryo is defined before formation of the primitive streak, and axis specification and subsequent anterior development involves signaling from both embryonic ectoderm and visceral endoderm. Tauhe Wnt signaling pathway is essential for various developmental processes, but a role in anterior-posterior axis formation in the mouse has not been previously established. Beta-catenin is a central player in the Wnt pathway and in cadherin-mediated cell adhesion. We generated beta-catenin-deficient mouse embryos and observed a defect in anterior-posterior axis formation at embryonic day 5.5, as visualized by the absence of Hex and Hesx1 and the mislocation of cerberus-like and Lim1 expression. Subsequently, no mesoderm and head structures are generated. Intercellular adhesion is maintained since plakoglobin substitutes for beta-catenin. Our data demonstrate that beta-catenin function is essential in anterior-posterior axis formation in the mouse, and experiments with chimeric embryos show that this function is required..

A novel interaction between ATOH8 and PPP3CB

ATOH8 is a bHLH transcription factor playing roles in a variety of developmental processes such as neurogenesis, differentiation of pancreatic precursor cells, development of kidney and muscle, and differentiation of endothelial cells. PPP3CB belongs to the catalytic subunit of the serine/threonine phosphatase, calcineurin, which can dephosphorylate its substrate proteins to regulate their physiological activities. In our study, we demonstrated that ATOH8 interacts with PPP3CB in vitro with different approaches. We show that the conserved catalytic domain of PPP3CB interacts with both the N-terminus and the bHLH domain of ATOH8. Although the interaction domain of PPP3CB is conserved among all isoforms of calcineurin A, ATOH8 selectively interacts with PPP3CB instead of PPP3CA, probably due to the unique proline-rich region present in the N-terminus of PPP3CB, which controls the specificity of its interaction partners. Furthermore, we show that inhibition of the interaction with calcineurin inhibitor, cyclosporin A (CsA), leads to the retention of ATOH8 to the cytoplasm, suggesting that the interaction renders nuclear localization of ATOH8 which may be critical to control its activity as transcription factor