Modulation of sterol homeostasis by the Cdc42p effectors Cla4p and Ste20p in the yeast Saccharomyces cerevisiae

This article is available open access through the publisher’s website at the link below. Copyright @ 2009 The Authors.The conserved Rho-type GTPase Cdc42p is a key regulator of signal transduction and polarity in eukaryotic cells. In the yeast Saccharomyces cerevisiae, Cdc42p promotes polarized growth through the p21-activated kinases Ste20p and Cla4p. Previously, we demonstrated that Ste20p forms a complex with Erg4p, Cbr1p and Ncp1p, which all catalyze important steps in sterol biosynthesis. CLA4 interacts genetically with ERG4 and NCP1. Furthermore, Erg4p, Ncp1p and Cbr1p play important roles in cell polarization during vegetative growth, mating and filamentation. As Ste20p and Cla4p are involved in these processes it seems likely that sterol biosynthetic enzymes and p21-activated kinases act in related pathways. Here, we demonstrate that the deletion of either STE20 or CLA4 results in increased levels of sterols. In addition, higher concentrations of steryl esters, the storage form of sterols, were observed in cla4Δ cells. CLA4 expression from a multicopy plasmid reduces enzyme activity of Are2p, the major steryl ester synthase, under aerobic conditions. Altogether, our data suggest that Ste20p and Cla4p may function as negative modulators of sterol biosynthesis. Moreover, Cla4p has a negative effect on steryl ester formation. As sterol homeostasis is crucial for cell polarization, Ste20p and Cla4p may regulate cell polarity in part through the modulation of sterol homeostasis.Deutsche Forschungsgemeinschaft and the Austrian FWF

Global MHD simulation of flux transfer events at the high-latitude magnetopause observed by the cluster spacecraft and the SuperDARN radar system

A global magnetohydrodynamic numerical simulation is used to study the large-scale structure and formation location of flux transfer events (FTEs) in synergy with in situ spacecraft and ground-based observations. During the main period of interest on the 14 February 2001 from 0930 to 1100 UT the Cluster spacecraft were approaching the Northern Hemisphere high-latitude magnetopause in the postnoon sector on an outbound trajectory. Throughout this period the magnetic field, electron, and ion sensors on board Cluster observed characteristic signatures of FTEs. A few minutes delayed to these observations the Super Dual Auroral Radar Network (SuperDARN) system indicated flow disturbances in the conjugate ionospheres. These “two-point” observations on the ground and in space were closely correlated and were caused by ongoing unsteady reconnection in the vicinity of the spacecraft. The three-dimensional structures and dynamics of the observed FTEs and the associated reconnection sites are studied by using the Block-Adaptive-Tree-Solarwind-Roe-Upwind-Scheme (BATS-R-US) MHD code in combination with a simple open flux tube motion model (Cooling). Using these two models the spatial and temporal evolution of the FTEs is estimated. The models fill the gaps left by measurements and allow a “point-to-point” mapping between the instruments in order to investigate the global structure of the phenomenon. The modeled results presented are in good correlation with previous theoretical and observational studies addressing individual features of FTEs

Extended microsatellite analysis in microsatellite stable, MSH2 and MLH1 mutation-negative HNPCC patients: Genetic reclassification and correlation with clinical features

Background: Hereditary nonpolyposis colorectal cancer (HNPCC) is an autosomal dominant disorder predisposing to predominantly colorectal cancer (CRC) and endometrial cancer frequently due to germline mutations in DNA mismatch repair (MMR) genes, mainly MLH1, MSH2 and also MSH6 in families seen to demonstrate an excess of endometrial cancer. As a consequence, tumors in HNPCC reveal alterations in the length of simple repetitive genomic sequences like poly-A, poly-T, CA or GT repeats (microsatellites) in at least 90% of the cases. Aim of the Study: The study cohort consisted of 25 HNPCC index patients ( 19 Amsterdam positive, 6 Bethesda positive) who revealed a microsatellite stable (MSS) - or low instable (MSI-L) - tumor phenotype with negative mutation analysis for the MMR genes MLH1 and MSH2. An extended marker panel (BAT40, D10S197, D13S153, D18S58, MYCL1) was analyzed for the tumors of these patients with regard to three aspects. First, to reconfirm the MSI-L phenotype found by the standard panel; second, to find minor MSIs which might point towards an MSH6 mutation, and third, to reconfirm the MSS status of hereditary tumors. The reconfirmation of the MSS status of tumors not caused by mutations in the MMR genes should allow one to define another entity of hereditary CRC. Their clinical features were compared with those of 150 patients with sporadic CRCs. Results: In this way, 17 MSS and 8 MSI-L tumors were reclassified as 5 MSS, 18 MSI-L and even 2 MSI-H ( high instability) tumors, the last being seen to demonstrate at least 4 instable markers out of 10. Among all family members, 87 malignancies were documented. The mean age of onset for CRCs was the lowest in the MSI-H-phenotyped patients with 40.5 +/- 4.9 years (vs. 47.0 +/- 14.6 and 49.8 +/- 11.9 years in MSI-L- and MSS-phenotyped patients, respectively). The percentage of CRC was the highest in families with MSS-phenotyped tumors (88%), followed by MSI-L-phenotyped ( 78%) and then by MSI-H-phenotyped (67%) tumors. MSS tumors were preferentially localized in the distal colon supposing a similar biologic behavior like sporadic CRC. MSH6 mutation analysis for the MSI-L and MSI-H patients revealed one truncating mutation for a patient initially with an MSS tumor, which was reclassified as MSI-L by analyzing the extended marker panel. Conclusion: Extended microsatellite analysis serves to evaluate the sensitivity of the reference panel for HNPCC detection and permits phenotype confirmation or upgrading. Additionally, it confirms the MSS status of hereditary CRCs not caused by the common mutations in the MMR genes and provides hints to another entity of hereditary CRC. Copyright (C) 2004 S. Karger AG, Basel

The Semiclassical Limit of Causal Dynamical Triangulations

Previous work has shown that the macroscopic structure of the theory of quantum gravity defined by causal dynamical triangulations (CDT) is compatible with that of a de Sitter universe. After emphasizing the strictly nonperturbative nature of this semiclassical limit we present a detailed study of the three-volume data, which allows us to re-confirm the de Sitter structure, exhibit short-distance discretization effects, and make a first detailed investigation of the presence of higher-order curvature terms in the effective action for the scale factor. Technically, we make use of a novel way of fixing the total four-volume in the simulations.Comment: 30 pages, 10 figure

Supramolecular organization of the human N-BAR domain in shaping the sarcolemma membrane

This is the final version of the article. Available from Elsevier via the DOI in this record.The 30 kDa N-BAR domain of the human Bin1 protein is essential for the generation of skeletal muscle T-tubules. By electron cryo-microscopy and electron cryo-tomography with a direct electron detector, we found that Bin1-N-BAR domains assemble into scaffolds of low long-range order that form flexible membrane tubules. The diameter of the tubules closely matches the curved shape of the N-BAR domain, which depends on the composition of the target membrane. These insights are fundamental to our understanding of T-tubule formation and function in human skeletal muscle.This work was supported by grants from the Deutsche Forschungsgemeinschaft (GRK 1026, SFB610) (A.A., T.G., J.B.), the BMBF ZIK program (A.M., J.B.), the European Regional Development Fund of the European Commission (A.M., T.G.: EFRE 1241 12 0001), and the state Sachsen-Anhalt (A.M., T.G., J.B.)

The influence of the geomagnetic field and of the uncertainties in the primary spectrum on the development of the muon flux in the atmosphere

In this paper we study the sensitivity of the flux of atmospheric muons to uncertainties in the primary cosmic ray spectrum and to the treatment of the geomagnetic field in a calculation. We use the air shower simulation program AIRES to make the calculation for two different primary spectra and under several approximations to the propagation of charged particles in the geomagnetic field. The results illustrate the importance of accurate modelling of the geomagnetic field effects. We propose a high and a low fit of the proton and helium fluxes, and calculate the muon fluxes with these different inputs. Comparison with measurements of the muon flux by the CAPRICE experiment shows a slight preference for the higher primary cosmic ray flux parametrization.Comment: 24 pages, 13 figures, submitted to Phys.Rev.

Comparison of ultracold neutron sources for fundamental physics measurements

Ultracold neutrons (UCNs) are key for precision studies of fundamental parameters of the neutron and in searches for new CP violating processes or exotic interactions beyond the Standard Model of particle physics. The most prominent example is the search for a permanent electric dipole moment of the neutron (nEDM). We have performed an experimental comparison of the leading UCN sources currently operating. We have used a 'standard' UCN storage bottle with a volume of 32 liters, comparable in size to nEDM experiments, which allows us to compare the UCN density available at a given beam port.Comment: 20 pages, 30 Figure

Visualization of ATP synthase dimers in mitochondria by electron cryo-tomography

This is the final version of the article. Available from the publisher via the DOI in this recordThe video component of this article can be found at http://www.jove.com/video/51228/Electron cryo-tomography is a powerful tool in structural biology, capable of visualizing the three-dimensional structure of biological samples, such as cells, organelles, membrane vesicles, or viruses at molecular detail. To achieve this, the aqueous sample is rapidly vitrified in liquid ethane, which preserves it in a close-to-native, frozen-hydrated state. In the electron microscope, tilt series are recorded at liquid nitrogen temperature, from which 3D tomograms are reconstructed. The signal-to-noise ratio of the tomographic volume is inherently low. Recognizable, recurring features are enhanced by subtomogram averaging, by which individual subvolumes are cut out, aligned and averaged to reduce noise. In this way, 3D maps with a resolution of 2 nm or better can be obtained. A fit of available high-resolution structures to the 3D volume then produces atomic models of protein complexes in their native environment. Here we show how we use electron cryo-tomography to study the in situ organization of large membrane protein complexes in mitochondria. We find that ATP synthases are organized in rows of dimers along highly curved apices of the inner membrane cristae, whereas complex I is randomly distributed in the membrane regions on either side of the rows. By subtomogram averaging we obtained a structure of the mitochondrial ATP synthase dimer within the cristae membrane.This work was supported by the Max Planck Society (K.M.D., B.D., A.W.M., T.B., T.B.B., D.J.M., and W.K.), the Cluster of Excellence Frankfurt “Macromolecular Complexes” funded by the Deutsche Forschungsgemeinschaft (W.K.) and a postdoctoral EMBO Long-Term Fellowship (V.A.M.G.)

A National Survey of Skin Infections, Care Behaviors and MRSA Knowledge in the United States

A nationally representative sample of approximately 2000 individuals was surveyed to assess SSTI infections over their lifetime and then prospectively over six-months. Knowledge of MRSA, future likelihood to self-treat a SSTI and self-care behaviors was also queried. Chi square tests, linear and multinomial regression were used for analysis. About 50% of those with a reported history of a SSTI typical of MRSA had sought medical treatment. MRSA knowledge was low: 28% of respondents could describe MRSA. Use of protective self-care behaviors that may reduce transmission, such as covering a lesion, differed with knowledge of MRSA and socio-demographics. Those reporting a history of a MRSA-like SSTI were more likely to respond that they would self-treat than those without such a history (OR 2.05 95% CI 1.40, 3.01; p<0.001). Since half of respondents reported not seeking care for past lesions, incidence determined from clinical encounters would greatly underestimate true incidence. MRSA knowledge was not associated with seeking medical care, but was associated with self-care practices that may decrease transmission.</p