ALBERT

Beschreibung: Seismic tomography images of the upper mantle structures beneath the Pacific Northwestern United States display a maze of high-velocity anomalies, many of which produce distorted waveforms evident in the USArray observations indicative of the Juan de Fuca (JdF) slab. The inferred location of the slab agrees quite well with existing contour lines defining the slab's upper interface. Synthetic waveforms generated from a recent tomography image fit teleseismic travel times quite well and also some of the waveform distortions. Regional earthquake data, however, require substantial changes to the tomographic velocities. By modeling regional waveforms of the 2008 Nevada earthquake, we find that the uppermost mantle of the 1D reference model AK135, the reference velocity model used for most tomographic studies, is too fast for the western United States. Here, we replace AK135 with mT7, a modification of an older Basin-and-Range model T7. We present two hybrid velocity structures satisfying the waveform data based on modified tomographic images and conventional slab wisdom. We derive P and SH velocity structures down to 660 km along two cross sections through the JdF slab. Our results indicate that the JdF slab is subducted to a depth of 250 km beneath the Seattle region, and terminates at a shallower depth beneath Portland region of Oregon to the south. The slab is about 60 km thick and has a P velocity increase of 5% with respect to mT7. In order to fit waveform complexities of teleseismic Gulf of Mexico and South American events, a slab-like high-velocity anomaly with velocity increases of 3% for P and 7% for SH is inferred just above the 660 discontinuity beneath Nevada.

Beschreibung: The 2010 eruptive activity at the Eyjafjallajökull volcanic system began 20 March with a basaltic flank eruption on a 300 m long fissure on the Fimmvörðuháls Pass, in between Eyjafjallajökull and Mýrdalsjökull volcanoes. The magma expelled from the fissure is olivine- and plagioclase-bearing mildly alkali basalt that exhibits uniform and rather primitive whole-rock composition. This event provides a rare opportunity to assess deep magmatic processes in Iceland. Melt inclusions (MIs) hosted in olivine phenocrysts were analyzed for their major, trace and volatile element concentrations to enable identification of magmatic source(s) for Eyjafjallajökull volcano and to better constrain processes occurring at depth. The MIs, in particular those in Mg-rich olivines, record primary magma composition before homogenisation and differentiation during magma ascent. The olivine phenocrysts hosting the MIs have a large compositional range, extending from Fo73 to Fo87, reflecting changes in the magma characteristics from the source to the surface. The MI compositions exhibit significant variations with MgO ranging from 5.2 to 7.2 wt%. This compositional range was caused by a binary mixing of two basaltic end-members followed by fractional crystallization process. The sources of these end-members are identical to those of Katla and Surtsey basalts, with a dominant role of the Katla source. Trace element characteristics of the Fimmvörðuháls MIs suggest important proportions of recycled oceanic crust in their mantle sources.

Beschreibung: Small-magnitude earthquakes and ground deformation are the precursors most frequently recorded before volcanic eruptions. Analogous signals (using acoustic emissions) have also been reported before the bulk brittle failure of crustal rock in the laboratory. Models based on laboratory and field data have focused on precursory behavior during deformation under a constant stress. A new model is proposed for extending analyses to deformation under an increasing stress. It describes how precursory time series can be determined from a parent relation between fracturing and stress, together with time-dependent changes in applied stress and rock resistance. The model applies to rock in which these stresses do not interact with each other and occupy volumes much smaller than the total volume being deformed. It identifies how the amounts of fracturing observed during deformation are controlled not only by stress concentrations at macroscopic heterogeneities, such as crack tips but also by rock composition, temperature, confining pressure, and the distribution of energy among atoms. The results appear to be scale independent, and so may be used to investigate whether the approach to bulk failure is limited by changes in applied stress or in rock weakening. When applied to pre-eruptive data from Hawaii, the analysis suggests that precursory signals are controlled by an increase in applied stress, rather than by creep deformation under a constant stress.

Beschreibung: The Colorado Plateau is a physiographic province in the western US with an average elevation of ∼1.9 km where, in contrast to neighboring provinces, there is little evidence of large scale tectonic deformation or magmatism. Recent availability of Earthscope/USArray seismic data allow us to better examine the crust and upper mantle structure beneath the region and test proposed explanations for the plateau's uplift and relative stability. Using phase velocities for fundamental mode Rayleigh waves and P receiver functions, we perform over 800 joint inversions for 1-D shear wave velocity VS profiles sampling the plateau and surrounding regions down to 150 km depth. We image a sharp change in crustal thickness at the western edge of the Colorado Plateau with a more gradual increase eastward moving into the Rocky Mountains. A relatively thick (≳100 km) lithosphere beneath the plateau extends into the Rocky Mountains to the north. We use empirical scaling relations to estimate densities from our VS results, and predict the associated gravity anomalies, which are inconsistent with the observed distribution of the Bouguer gravity anomalies. We somewhat reconcile the prediction and observations by assuming that lateral density variations below 50 km can be ignored and the lithospheric root is therefore neutrally buoyant. While there is some evidence for small scale convection and lithospheric removal at its edges, the shape of the lithospheric mantle anomaly is consistent with a large scale uplift of the plateau by heating since removal of the Farallon slab. We conclude that the lithospheric root is key to the long term stability of the Colorado Plateau, leading to a colder, stronger crust.

Beschreibung: Hysteresis loops provide essential information concerning both induced and remanent magnetizations and are an important tool for characterizing magnetic mineral assemblages. Although the hysteresis behavior of mixed natural magnetic assemblages has been a focal point of much recent work, little progress has been made in unmixing of hysteresis loops into characteristic components. Unmixing strategies can act as cornerstones for interpretation of rock magnetic data and have become popular for characterizing isothermal remanent magnetization acquisition curves. Unmixing of hysteresis loops is, however, a challenging task because the individual component loops in the mixture must meet stringent shape constraints. We present a new technique for decomposing an ensemble of hysteresis loops into a small number of end-members based on linear mixing theory. The end-members are not based on type curves but instead are derived directly from the hysteresis data. Particular attention is paid to the form of the end-members, ensuring they meet the shape constraints expected for hysteresis loops of natural magnetic mineral assemblages. Marine sediments from the Southern Ocean and lake sediments from Butte Valley, northern California, provide case studies on which the proposed unmixing method is tested.

Beschreibung: This paper presents a new constitutive model that simulates the mechanical behavior of methane hydrate-bearing soil based on the concept of critical state soil mechanics, referred to as the “Methane Hydrate Critical State (MHCS) model”. Methane hydrate-bearing soil is, under certain geological conditions, known to exhibit greater stiffness, strength and dilatancy, which are often observed in dense soils and also in bonded soils such as cemented soil and unsaturated soil. Those soils tend to show greater resistance to compressive deformation but the tendency disappears when the soil is excessively compressed or the bonds are destroyed due to shearing. The proposed model represents these features by introducing five extra model parameters to the conventional critical state model. It is found that, for an accurate prediction of ground settlement, volumetric yielding plays an important role when hydrate soil undergoes a significant change in effective stresses and hydrate saturation, which are expected during depressurization for methane gas recovery.

Beschreibung: Experimental heating tests were performed on Volterra gypsum to study the micromechanical consequences of the dehydration reaction. The experimental conditions were drained at 5 MPa fluid pressure and confining pressures ranging from 15 to 55 MPa. One test was performed with a constantly applied differential stress of 30 MPa. The reaction is marked by (1) a porosity increase and homogeneous compaction, (2) a swarm of acoustic emissions, (3) a large decrease in P and S wave velocities, and (4) a decrease in VP/VS ratio. Wave velocity data are interpreted in terms of crack density and pore aspect ratio, which, modeling pores as spheroids, is estimated at around 0.05 (crack-like spheroid). Complementary tests performed in an environmental scanning electron microscope indicate that cracks first form inside the gypsum grains and are oriented preferentially along the crystal structure of gypsum. Most of the visible porosity appears at later stages when grains shrink and grain boundaries open. Extrapolation of our data to serpentinites in subduction zones suggest that the signature of dehydrating rocks in seismic tomography could be a low apparent Poisson's ratio, although this interpretation may be masked by anisotropy development due to preexisting crystal preferred orientation and/or deformation-induced cracking. The large compaction and the absence of strain localization in the deformation test suggests that dehydrating rocks maybe seen as soft inclusions and could thus induce ruptures in the surrounding, nonreacting rocks.

Beschreibung: We use a two-dimensional displacement discontinuity method (DDM) for quasi-static boundary value problems to investigate sinusoidal faults of finite length in an otherwise homogeneous and isotropic elastic material. The DDM incorporates a complementarity algorithm to enforce appropriate contact boundary conditions along the model fault. The numerical solution for the model sinusoidal fault converges to the analytical solution for a straight fault of finite length as the ratio amplitude/wavelength goes to zero. It does not converge to the analytical solution for an infinite sinusoidal interface as the ratio distance/wavelength goes to zero. We provide stick, slip, and opening distributions along wavy faults with a range of uniform coefficients of friction, amplitude/wavelength ratios, and wave numbers. As the number of sinusoidal waves or the amplitude/wavelength is increased, mean slip decreases. Additionally, the fault geometry causes slip to deviate significantly from the elliptical distribution of a planar fault. We demonstrate that the displacement discontinuity of wavy faults cannot be prescribed a priori. This necessitates implementation of the complementarity algorithm and precludes an analytical solution. We employ the terms lee and stoss instead of releasing and restraining bends because a local minimum in slip may occur along lee sides, as well as stoss sides. In some cases, lee sides stick while stoss sides slip. Trends in the slip perturbation can be explained by the angular relationship between the local fault trace and the orientation of the remote principal stresses; however, the displacement discontinuity along a wavy model fault cannot be explained by this relationship alone.

Beschreibung: At Cranfield, Mississippi, United States, a large-scale carbon dioxide (CO2) injection through an injection well (∼3,080 m deep) was continuously monitored using U-tube samplers in two observation wells located 68 and 112 m east of the injector. The Lower Tuscaloosa Formation injection zone, which consists of amalgamated fluvial point-bar and channel-fill deposits, presents an interesting environment for studying fluid flow in heterogeneous formations. Continual fluid sampling was carried out during the first month of CO2 injection. Two subsequent tracer tests using sulfur hexafluoride (SF6) and krypton were conducted at different injection rates to measure flow velocity change. The field observations showed significant heterogeneity of fluid flow and for the first time clearly demonstrated that fluid flow evolved with time and injection rate. It was found the wells were connected through numerous, separate flow pathways. CO2 flowed through an increasing fraction of the reservoir and sweep efficiency improved with time. The field study also first documented in situ component exchange between brine and gas phases during CO2 injection. It was found that CH4 degassed from brine and is enriched along the gas–water contact. Multiple injectate flow fronts with high CH4 concentration arrived at different times and led to gas composition fluctuations in the observation wells. The findings provide valuable insights into heterogeneous multiphase flow in rock formations and show that conventional geological models and static fluid flow simulations are unable to fully describe the heterogeneous and dynamic flow during fluid injection.

Beschreibung: By regressing simple, independent variables that describe climate and tectonic processes against measures of topography and relief of 69 mountain ranges worldwide, we quantify the relative importance of these processes in shaping observed landscapes. Climate variables include latitude (as a surrogate for mean annual temperature and insolation, but most importantly for the likelihood of glaciation) and mean annual precipitation. To quantify tectonics we use shortening rates across each range. As a measure of topography, we use mean and maximum elevations and relief calculated over different length scales. We show that the combination of climate (negative correlation) and tectonics (positive correlation) explain substantial fractions (〉25%, but

Beschreibung: We present a new approach to extracting spatially and temporally continuous ground deformation fields from interferometric synthetic aperture radar (InSAR) data. We focus on unwrapped interferograms from a single viewing geometry, estimating ground deformation along the line-of-sight. Our approach is based on a wavelet decomposition in space and a general parametrization in time. We refer to this approach as MInTS (Multiscale InSAR Time Series). The wavelet decomposition efficiently deals with commonly seen spatial covariances in repeat-pass InSAR measurements, since the coefficients of the wavelets are essentially spatially uncorrelated. Our time-dependent parametrization is capable of capturing both recognized and unrecognized processes, and is not arbitrarily tied to the times of the SAR acquisitions. We estimate deformation in the wavelet-domain, using a cross-validated, regularized least squares inversion. We include a model-resolution-based regularization, in order to more heavily damp the model during periods of sparse SAR acquisitions, compared to during times of dense acquisitions. To illustrate the application of MInTS, we consider a catalog of 92 ERS and Envisat interferograms, spanning 16 years, in the Long Valley caldera, CA, region. MInTS analysis captures the ground deformation with high spatial density over the Long Valley region.

Beschreibung: In shallow magmatic intrusions, a characteristic layering structure (hereafter referred to as cyclic layering) can sometimes be observed. This cyclic layering is caused by double diffusion and crystallization kinetics, and different from what is observed as rhythmic layering caused by gravity. The cyclic layering is visualized as differential weathering in response to the differential stiffness caused by textural variations such as those in the volume fraction, number density, and size of vesicles or crystals. The spacing of layers seems to increase according to a geometric progression, like as in Liesegang bands of a diffusion-precipitation system. In order to understand the development condition for cyclic layering and the characteristics of textural variations, such as the spacing of layering in crystallized multi-component melts by conductive cooling, we carried out a numerical experiment on the 1D crystallization process of a binary eutectic melt. This simulation took into account the cooling from contact with country rock as well as the compositional and thermal diffusion and the kinetics of diffusion-limited crystallization. The governing equations include dimensionless control parameters describing the relative importance of thermal diffusion or compositional diffusion (Lewis number, Le) and the effective latent heat release (Stefan number, St). From the results of the numerical experiments, it was found that the layering develops through eutectic oscillation (compositional and thermal oscillation below the eutectic point), suggesting that the bi-activating condition, whereby both phases cooperatively activate their crystallization rates, is essential for the development of layering. No layering is observed at the margin, and the length of the region with no layering increases exponentially with decreasing St. The amplitude of textural oscillation decreases with decreasing St. Thus, practically no layering develops at small latent heat release. Three types of layering structure or oscillatory profiles of texture are observed (short, long and multiple types), depending mainly on Le. Realistic values of Le and St suggest that natural cyclic layering is the multiple or long type of layering. The common ratios of geometric progressions converge with increasing Le to constants in the range of approximately 1.02–1.05, which is similar to the range of the natural observations. Experiments with no latent heat release by the second-phase simulating vesicles show similar oscillatory behaviors, suggesting that the latent heat release of the first crystallizing phase is an essential factor for the development of vesicle layering.

Beschreibung: 27 December 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 6 Insulin stimulates glucose uptake through the membrane translocation of GLUT4 and GLUT1. Peroxisome proliferator-activated receptor γ (PPARγ) enhances insulin sensitivity. Here, we demonstrate that insulin stimulates GLUT4 and GLUT1 translocation, and glucose uptake, by activating the signaling pathway involving nicotinic acid adenine dinucleotide phosphate (NAADP), a calcium mobilizer, in adipocytes. We also demonstrate that PPARγ mediates insulin sensitization by enhancing NAADP production through upregulation of CD38, the only enzyme identified for NAADP synthesis. Insulin produced NAADP by both CD38-dependent and -independent pathways, whereas PPARγ produced NAADP by CD38-dependent pathway. Blocking the NAADP signaling pathway abrogated both insulin-stimulated and PPARγ-induced GLUT4 and GLUT1 translocation, thereby inhibiting glucose uptake. CD38 knockout partially inhibited insulin-stimulated glucose uptake. However, CD38 knockout completely blocked PPARγ-induced glucose uptake in adipocytes and PPARγ-mediated amelioration of glucose tolerance in diabetic mice. These results demonstrated that the NAADP signaling pathway is a critical molecular target for PPARγ-mediated insulin sensitization. Graphical abstract Highlights ► Insulin induces glucose uptake via NAADP-mediated calcium increase ► PPARγ mediates insulin sensitization by upregulating the NAADP-producing enzyme CD38 ► PPARγ agonists ameliorate glucose tolerance by CD38 upregulation

Beschreibung: Available online 27 December 2012 Publication year: 2012 Source: Cell Reports Enveloped viruses have developed various adroit mechanisms to invade their host cells. This process requires one or more viral envelope glycoprotein to achieve cell attachment and membrane fusion. Members of the Flaviviridae such as flaviviruses possess only one envelope glycoprotein, E, whereas pestiviruses and hepacivirus encode two glycoproteins, E1 and E2. Although E2 is involved in cell attachment, it has been unclear which protein is responsible for membrane fusion. We report the crystal structures of the homodimeric glycoprotein E2 from the pestivirus bovine viral diarrhea virus 1 (BVDV1) at both neutral and low pH. Unexpectedly, BVDV1 E2 does not have a class II fusion protein fold, and at low pH the N-terminal domain is disordered, similarly to the intermediate postfusion state of E2 from sindbis virus, an alphavirus. Our results suggest that the pestivirus and possibly the hepacivirus fusion machinery are unlike any previously observed. Graphical abstract Highlights ► Structure of the major antigenically dominant protein of BVDV ► The overall fold of BVDV E2 shows no similarity to the class II fusion proteins ► At low pH, BVDV E2 N-terminal domain is disordered ► Entry mechanism of BVDV is probably applicable to hepatitis C virus

Beschreibung: 27 December 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 6 Controversy has recently arisen over the role of sirtuins in metazoan aging. In this issue of Cell Reports , Banerjee et al. demonstrate that Drosophila Sir2 is necessary for life span extension in response to dietary restriction and that its overexpression in the fat body increases the life span.

Beschreibung: 27 December 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 6 Inhibition of sirtuin 2 (SIRT2) deacetylase mediates protective effects in cell and invertebrate models of Parkinson’s disease and Huntington’s disease (HD). Here we report the in vivo efficacy of a brain-permeable SIRT2 inhibitor in two genetic mouse models of HD. Compound treatment resulted in improved motor function, extended survival, and reduced brain atrophy and is associated with marked reduction of aggregated mutant huntingtin, a hallmark of HD pathology. Our results provide preclinical validation of SIRT2 inhibition as a potential therapeutic target for HD and support the further development of SIRT2 inhibitors for testing in humans. Graphical abstract Highlights ► Sirtuin 2 (SIRT2) inhibition is neuroprotective in two HD mouse models ► SIRT2 inhibitor treatment markedly reduces huntingtin aggregates in HD mouse brain ► SIRT2 is a promising therapeutic target for neurological protein aggregation disorders

Beschreibung: 27 December 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 6 Planar cell polarity (PCP) refers to the collective orientation of cells within the epithelial plane. We show that progenitor cells forming the ducts of the embryonic pancreas express PCP proteins and exhibit an active PCP pathway. Planar polarity proteins are acquired at embryonic day 11.5 synchronously to apicobasal polarization of pancreas progenitors. Loss of function of the two PCP core components Celsr2 and Celsr3 shows that they control the differentiation of endocrine cells from polarized progenitors, with a prevalent effect on insulin-producing beta cells. This results in a decreased glucose clearance. Loss of Celsr2 and 3 leads to a reduction of Jun phosphorylation in progenitors, which, in turn, reduces beta cell differentiation from endocrine progenitors. These results highlight the importance of the PCP pathway in cell differentiation in vertebrates. In addition, they reveal that tridimensional organization and collective communication of cells are needed in the pancreatic epithelium in order to generate appropriate numbers of endocrine cells. Graphical abstract Highlights ► PCP proteins are restricted to progenitor cells in the embryonic pancreas ► The PCP core components Celsr2 and Celsr3 control endocrine cell differentiation ► This effect is mediated by the JNK pathway

Beschreibung: 27 December 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 6 MicroRNAs (miRNAs) are small, noncoding RNAs that inhibit translation and promote mRNA decay. The levels of mature miRNAs are the result of different rates of transcription, processing, and turnover. The noncanonical polymerase Gld2 has been implicated in the stabilization of miR-122, possibly through catalyzing 3′ monoadenylation; however, there is little evidence that this relationship is one of cause and effect. Here, we biochemically characterize Gld2’s involvement in miRNA monoadenylation and its effect on miRNA stability. We find that Gld2 directly monoadenylates and stabilizes specific miRNA populations in human fibroblasts and that sensitivity to monoadenylation-induced stability depends on nucleotides in the miRNA 3′ end. These results establish a mechanism of miRNA stability and resulting posttranscriptional gene regulation. Graphical abstract Highlights ► Gld2 monoadenylates specific microRNAs (miRNAs) ► Monoadenylation stabilizes miRNA subpopulations and prolongs their activity ► Sensitivity to monoadenylation and stability depends on nucleotides in the miRNA 3′ end

Beschreibung: 27 December 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 6 In the cytoplasm, the correct delivery of membrane proteins is an essential and highly regulated process. The posttranslational targeting of the important tail-anchor membrane (TA) proteins has recently been under intense investigation. A specialized pathway, called the guided entry of TA proteins (GET) pathway in yeast and the transmembrane domain recognition complex (TRC) pathway in vertebrates, recognizes endoplasmic-reticulum-targeted TA proteins and delivers them through a complex series of handoffs. An early step is the formation of a complex between Sgt2/SGTA, a cochaperone with a presumed ubiquitin-like-binding domain (UBD), and Get5/UBL4A, a ubiquitin-like domain (UBL)-containing protein. We structurally characterize this UBD/UBL interaction for both yeast and human proteins. This characterization is supported by biophysical studies that demonstrate that complex formation is mediated by electrostatics, generating an interface that has high-affinity with rapid kinetics. In total, this work provides a refined model of the interplay of Sgt2 homologs in TA targeting. Graphical abstract Highlights ► Sgt2 homologs contain dimerization motifs that are novel UBDs ► The UBL of Get5 homologs has features that distinguish it from other UBLs ► Complex formation of Sgt2 and Get5 includes a conserved and dynamic interface ► The system can rapidly discriminate between other UBL pathways

Beschreibung: 27 December 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 6 Chimera formation after blastocyst injection or morula aggregation is the principal functional assay of the developmental potential of mouse embryonic stem cells (ESCs). This property, which demonstrates functional equivalence between ESCs and the preimplantation epiblast, is not shared by epiblast stem cell (EpiSC) lines. Here, we show that EpiSCs derived either from postimplantation embryos or from ESCs in vitro readily generate chimeras when grafted to postimplantation embryos in whole embryo culture. EpiSC derivatives integrate and differentiate to derivatives of all three embryonic germ layers and primordial germ cells. In contrast, grafted ESCs seldom proliferate in postimplantation embryos, and fail to acquire the identity of their host-derived neighbors. EpiSCs do not incorporate efficiently into embryonic day 8.5 embryos, a stage by which pluripotency has been lost. Thus, chimera formation by EpiSCs requires a permissive environment, the postimplantation epiblast, and demonstrates functional equivalence between this cell type and EpiSCs. Graphical abstract Highlights ► Epiblast stem cells (EpiSCs) form chimeras when injected into postimplantation epiblast ► Embryonic stem cells do not form postimplantation chimeras ► EpiSCs do not integrate if they are injected after gastrulation

Beschreibung: 27 December 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 6 An increased understanding of antitumor immunity is necessary for improving cell-based immunotherapies against human cancers. Here, we investigated the roles of two immune system-expressed microRNAs (miRNAs), miR-155 and miR-146a, in the regulation of antitumor immune responses. Our results indicate that miR-155 promotes and miR-146a inhibits interferon γ (IFNγ) responses by T cells and reduces solid tumor growth in vivo. Using a double-knockout (DKO) mouse strain deficient in both miR-155 and miR-146a, we have also identified an epistatic relationship between these two miRNAs. DKO mice had defective T cell responses and tumor growth phenotypes similar to miR-155 −/− mice. Further analysis of the T cell compartment revealed that miR-155 modulates IFNγ expression through a mechanism involving repression of Ship1. Our work reveals critical roles for miRNAs in the reciprocal regulation of CD4 + and CD8 + T cell-mediated antitumor immunity and demonstrates the dominant nature of miR-155 during its promotion of immune responses. Graphical abstract Highlights ► miR-155 promotes and miR-146a inhibits both CD4+ and CD8+ T cell antitumor responses ► DKO mice reveal epistasis between miR-155 and miR-146a during tumor immunity ► miR-155 regulation of IFNγ involves repression of its target Ship1 in T cells

Beschreibung: Available online 20 December 2012 Publication year: 2012 Source: Cell Reports In the fission yeast Schizosaccharomyces pombe , a chromosomal imprinting event controls the asymmetric pattern of mating-type switching. The orientation of DNA replication at the mating-type locus is instrumental in this process. However, the factors leading to imprinting are not fully identified and the mechanism is poorly understood. Here, we show that the replication fork pause at the mat1 locus ( MPS1 ), essential for imprint formation, depends on the lysine-specific demethylase Lsd1. We demonstrate that either Lsd1 or Lsd2 amine oxidase activity is required for these processes, working upstream of the imprinting factors Swi1 and Swi3 (homologs of mammalian Timeless and Tipin, respectively). We also show that the Lsd1/2 complex controls the replication fork terminators, within the rDNA repeats. These findings reveal a role for the Lsd1/2 demethylases in controlling polar replication fork progression, imprint formation, and subsequent asymmetric cell divisions. Graphical abstract Highlights ► The lysine-specific demethylase, Lsd1, is a mating-type switching mutant ► Lsd1 is essential for replication fork pausing at the mat1 locus ► Lsd1 is essential for replication fork arrest at the replication fork block at rDNA loci ► Lsd1 and Lsd2 act in a redundant manner for replication fork pausing

Beschreibung: Available online 21 December 2012 Publication year: 2012 Source: Cell Reports Chromatin dynamics play a central role in maintaining genome integrity, but how this is achieved remains largely unknown. Here, we report that microrchidia CW-type zinc finger 2 (MORC2), an uncharacterized protein with a derived PHD finger domain and a conserved GHKL-type ATPase module, is a physiological substrate of p21-activated kinase 1 (PAK1), an important integrator of extracellular signals and nuclear processes. Following DNA damage, MORC2 is phosphorylated on serine 739 in a PAK1-dependent manner, and phosphorylated MORC2 regulates its DNA-dependent ATPase activity to facilitate chromatin remodeling. Moreover, MORC2 associates with chromatin and promotes gamma-H2AX induction in a PAK1 phosphorylation-dependent manner. Consequently, cells expressing MORC2-S739A mutation displayed a reduction in DNA repair efficiency and were hypersensitive to DNA-damaging agent. These findings suggest that the PAK1-MORC2 axis is critical for orchestrating the interplay between chromatin dynamics and the maintenance of genomic integrity through sequentially integrating multiple essential enzymatic processes. Graphical abstract Highlights ► MORC2 is a DNA damage-responsive phosphoprotein activated by PAK1 kinase ► MORC2 regulates phosphorylation-coupled, ATPase-dependent chromatin remodeling ► MORC2 facilitates gamma-H2AX induction independently of PIKK kinases ► MORC2 promotes DSB repair in a PAK1 phosphorylation-dependent manner

Beschreibung: Using fundamental mode Rayleigh waves from the INDEPTH-IV and Namche-Barwa seismic experiments for periods between 20 and 143 s, we have investigated the lithospheric structure beneath eastern Tibet. We have found a ∼200-km-wide high velocity body, starting at ∼60 km depth and roughly centered beneath the Bangong-Nijuang Suture, which is most likely a piece of the underthrusting Indian continental lithosphere. The sub-horizontal underthrusting of the Indian lithosphere beneath eastern Tibet appears to be accompanied by its lateral tearing into at least two fragments, and subsequent break-off of the westernmost portion at ∼91°E-33°N. The uppermost mantle low velocity zone we observe beneath the N. Qiangtang and Songpan-Ganzi terranes is most probably due to warmer and thinner lithosphere relative to southern Tibet. We attribute the low velocity zones concentrated along the northern and southern branches of the eastern Kunlun fault at lithospheric depths to strain heating caused by shearing. The azimuthal fast directions at all periods up to 143 s (∼200 km peak sensitivity depth) beneath the N. Qiangtang and Songpan-Ganzi terranes are consistent, suggesting vertically coherent deformation between crust and uppermost mantle. Furthermore, the low velocity zone below the Kunlun Shan reaching down to 〉200 km argues against a present southward continental subduction along the southern margin of Qaidam Basin.

Beschreibung: Publication year: 2012 Source: Cell Reports Hong Zan, Clayton A. White, Lisa M. Thomas, Thach Mai, Guideng Li, Zhenming Xu, Jinsong Zhang, Paolo Casali By diversifying the biological effector functions of antibodies, class switch DNA recombination (CSR) plays a critical role in the maturation of the immune response. It is initiated by activation-induced cytidine deaminase (AID)-mediated deoxycytosine deamination, yielding deoxyuridine (dU), and dU glycosylation by uracil DNA glycosylase (Ung) in antibody switch (S) region DNA. Here we showed that the translesion DNA synthesis polymerase Rev1 directly interacted with Ung and targeted in an AID-dependent and Ung-independent fashion the S regions undergoing CSR. Rev1 −/− Ung +/+ B cells reduced Ung recruitment to S regions, DNA-dU glycosylation, and CSR. Together with an S region spectrum of mutations similar to that of Rev1 +/+ Ung −/− B cells, this suggests that Rev1 operates in the same pathway as Ung, as emphasized by further decreased CSR in Rev1 −/− Msh2 −/− B cells. Rescue of CSR in Rev1 −/− B cells by a catalytically inactive Rev1 mutant shows that the important role of Rev1 in CSR is mediated by Rev1’s scaffolding function, not its enzymatic function. Graphical abstract Graphical Abstract Highlights ► Rev1 DNA polymerase plays an important role in class switch DNA recombination ► This role is mediated by Rev1’s scaffolding function, not its enzymatic function ► Rev1 interacts with Ung and recruits it to switch region DNA ► Rev1 enhances Ung-mediated dU glycosylation in DNA

Beschreibung: Publication year: 2012 Source: Cell Reports Michihiko Sugimoto, Masayo Kondo, Michiko Hirose, Misao Suzuki, Kazuyuki Mekada, Takaya Abe, Hiroshi Kiyonari, Atsuo Ogura, Nobuo Takagi, Karen Artzt, Kuniya Abe After implantation, pluripotent epiblasts are converted to embryonic ectoderm through cell–cell interactions that significantly change the transcriptional and epigenetic networks. An entrée to understanding this vital developmental transition is the t w5 mutation of the mouse t complex. This mutation produces highly specific defects in the embryonic ectoderm before gastrulation, leading to death of the embryonic ectoderm. Using a positional cloning approach, we have now identified the mutated gene, completing a decades-long search. The gene, vacuolar protein sorting 52 ( Vps52 ), is a mouse homolog of yeast VPS52 that is involved in the retrograde trafficking of endosomes. Our data suggest that Vps52 acts in extraembryonic tissues to support the growth and differentiation of embryonic ectoderm via cell–cell interactions. It is also required in the formation of embryonic structures at a later stage of development, revealing hitherto unknown functions of Vps52 in the development of a multicellular organism. Graphical abstract Graphical Abstract Highlights ► Vps52 , part of retrograde transport to the Golgi, causes the t w5 lethal phenotype ► Vps52 acts in extraembryonic tissues to dictate embryonic ectoderm differentiation ► VPS52’s essential function in the embryo involves cell–cell interactions ► VPS52 also has an indispensable function later in development

Beschreibung: Publication year: 2012 Source: Cell Reports Myrto Denaxa, Melanie Kalaitzidou, Anna Garefalaki, Angeliki Achimastou, Reena Lasrado, Tamara Maes, Vassilis Pachnis The generation of cortical interneuron subtypes is controlled by genetic programs that are activated in the ventral forebrain and unfold during the prolonged period of inhibitory neuron development. The LIM-homeodomain protein LHX6 is critical for the development of all cortical interneurons originating in the medial ganglionic eminence, but the molecular mechanisms that operate downstream of LHX6 to control the terminal differentiation of somatostatin- and parvalbumin-expressing interneurons within the cortex remain unknown. Here, we provide evidence that the nuclear matrix and genome organizer protein SATB1 is induced by neuronal activity and functions downstream of Lhx6 to control the transition of tangentially migrating immature interneurons into the terminally differentiated Somatostatin (SST)-expressing subtype. Our experiments provide a molecular framework for understanding the genetic and epigenetic mechanisms by which specified but immature cortical interneurons acquire the subtype-defining molecular and morphophysiological characteristics that allow them to integrate and function within cortical circuits. Graphical abstract Graphical Abstract Highlights ► Satb1 is specifically expressed in MGE-derived cortical interneurons ► Satb1 is required for the maturation of MGE-derived cortical interneurons in vivo ► Satb1 promotes the maturation of SST+ cortical interneurons downstream of Lhx6 ► Neuronal activity induces expression of Satb1 in immature cortical interneurons

Beschreibung: Available online 7 December 2012 Publication year: 2012 Source: Cell Reports Periodic somite segmentation is controlled by the cyclic gene Hes7 , whose oscillatory expression depends upon negative feedback with a delayed timing. The mechanism that regulates the pace of segmentation remains to be determined, but mathematical modeling has predicted that negative feedback with shorter delays would give rise to dampened but more rapid oscillations. Here, we show that reducing the number of introns within the Hes7 gene shortens the delay and results in a more rapid tempo of both Hes7 oscillation and somite segmentation, increasing the number of somites and vertebrae in the cervical and upper thoracic region. These results suggest that the number of introns is important for the appropriate tempo of oscillatory expression and that Hes7 is a key regulator of the pace of the segmentation clock. Graphical abstract Highlights ► Reduction of Hes7 intron number accelerates the tempo of Hes7 oscillation ► Reduction of Hes7 intron number accelerates the tempo of somite segmentation ► Hes7 is a key segmentation pacemaker ► The number of introns may be important for normal timing of gene expression

Beschreibung: The unique physical and chemical properties of cratonic lithosphere are thought to be key to its long-term survival and its resistance to pervasive modification by tectonic processes. Study of mantle structure in southeast Canada and the northeast US offers an excellent opportunity to address this issue because the region spans 3 billion years of Earth history, including Archean formation of the Superior craton and younger accretion of terranes to eastern Laurentia during the Proterozoic Grenville and Phanerozoic Appalachian orogenies. Trending NW–SE through each of these terranes is the track of the Great Meteor hot spot, which affected the region during the Mesozoic. Here we study mantle seismic velocity structure beneath this region of eastern North America using tomographic inversion of teleseismic P-wave relative arrival-times recorded by a large-aperture seismograph network. There are no large-scale systematic differences between Superior and Grenville mantle wave speed structure, which may suggest that tectonic stabilization of cratons occurred in a similar fashion during the Archean and Proterozoic. Cratonic lithosphere is largely thought to be resistant to modification by hot spot processes, in contrast to younger terranes where lithospheric erosion and significant magmatism are expected. Low velocities beneath the regions affected by the Great Meteor hot spot are broadest beneath the Paleozoic Appalachian terranes, indicating pervasive modification of the lithosphere during magmatism. The zone of modification narrows considerably into the Proterozoic Grenville province before disappearing completely in the Archean Superior craton, where the surface signature of Mesozoic magmatism is limited to kimberlite eruptions.

Beschreibung: It is well known that the Ionian Sea is characterized by thin (8–11 km) crystalline crust, thick (5–7 km) sedimentary cover, and low heat flow, typical for a Mesozoic (at least) basin. Yet seismic data have not yielded univocal interpretations, and a debate has developed on the oceanic versus “thinned continental” nature of the Ionian basin. Here we analyze the magnetic anomaly pattern of the Ionian Sea and compare it to synthetic fields produced by a geopotential field generator, considering realistic crust geometry. The Ionian basin is mostly characterized by slightly negative magnetic residuals and by a prominent positive (150 nT at sea level) “B” anomaly at the northwestern basin margin. We first test continental crust models, considering a homogeneous crystalline crust with K = 1 × 10−3, then a 5 km thick deep crustal layer of serpentinite (K = 1 × 10−1). The first model yields insignificant anomalies, while the second gives an anomaly pattern anticorrelated with the observed residuals. We subsequently test oceanic crust models, considering a 2 km thick 2A basaltic layer with K = 5 × 10−3, magnetic remanence of 5 A/m, and a unique magnetic polarity (no typical oceanic magnetic anomaly stripes are apparent in the observed data set). Magnetic remanence directions were derived from Pangean-African paleopoles in the 290–190 Ma age window. Only reverse polarity models reproduce the B anomaly, and among them the 220–230 Ma models best approximate magnetic features observed on the abyssal plain and at the western basin boundary. The Ionian Sea turns out to be the oldest preserved oceanic floor known so far.

Beschreibung: The devastation inflicted by recent earthquakes demonstrates the danger of under-predicting the size of earthquakes. Unfortunately, earthquakes may rupture fault-sections larger than previously observed, making it essential to develop predictive rupture models. We present numerical models based on earthquake physics and fault zone data, that determine whether a rupture on a segmented fault could cascade and grow into a devastating, multisegment earthquake. We demonstrate that weakened (damaged) fault zones and bi-material interfaces promote rupture propagation and greatly increase the risk of cascading ruptures and triggered seismicity. This result provides a feasible explanation for the outstanding observation of a very large (10 km) rupture jump documented in the MW7.8 2001 Kunlun, China earthquake. However, enhanced inter-seismic deformation and energy dissipation at fault tips suppress rupture propagation and may turn even small discontinuities into effective earthquake barriers. By assessing fault stability, identifying rupture barriers and foreseeing multisegment earthquakes, we provide a tool to improve earthquake prediction and hazard analysis.

Beschreibung: We use first-principles molecular dynamics simulations to study the behavior of cation ordering in the non-equivalent octahedral sites of Mg-Fe olivine solid solutions. Our theoretical calculations confirm the previous experimental finding that Mg2+ and Fe2+ can invert their octahedral site occupancy at a critical temperature. Assuming that the site preference of Fe changes discontinuously between two states in which it is completely restricted to either M1 or M2 sites, we have calculated the transition temperature, Tt, between the two extreme states. Under ambient pressure Tt is calculated to be 520°C that agrees fairly with the experimental finding in which, however, the ordering state changed discontinuously over a much smaller range of the site occupancy of Fe. Tt is found to be pressure sensitive, showing an increase by 30 to 100°C per unit GPa, depending upon the iron content. Using the Indian continental geotherm, we estimate a depth of around 75 Km corresponding to the calculated transition pressure and temperature of cation ordering, which matches well with the depth for the Hales discontinuity marked by a jump of shear wave velocity by ∼4%. For olivine solid solutions with 12.5% iron, the ordering transition increases Vs from 4.5 to 4.7 Km/s. Both the inferences, viz. depth of discontinuity and magnitude of velocity increase find support from the modeling of teleseismic earthquake waveforms recorded over broadband seismographs on the Dharwar Craton. This leads us to infer that the cation ordering transition in ferromagnesian olivine might be a potential factor for the Hales discontinuity.

Beschreibung: Deformation fabrics in Earth's crust and mantle are commonly used to constrain the tectonic history, deformation mechanisms, and rheological properties of the lithosphere. Their formation involves heterogeneous and multiscale deformation processes that current single-scale models cannot capture. Here we present a micromechanics-based MultiOrder Power Law Approach (MOPLA) for the simulation of multiscale fabrics in crustal scale high-strain zones. We consider the progressive deformation in a crustal high-strain zone on three different scales. On the macroscopic scale, representing the average assemblage of rock units at a point, we regard the rock mass as a continuum made of many first-order elements. The progressive deformation of first-order elements in the macroscopic flow field simulates tectonic transposition. On the scale of an individual first-order element, we regard it as an Eshelby inhomogeneity embedded in a poly element continuum. We apply Eshelby's inhomogeneity formalism for power law materials to relate the flow field inside a first-order element to the macroscopic flow field. On the scale pertinent to structures observed on the outcrop or smaller scale, the partitioned flow fields inside individual first-order elements are used to examine the fabric development. We implement MOPLA in MathCad, apply the approach to a natural example of the Cascade Lake shear zone, and discuss the implications of multiscale deformation. Our model predicts lineation patterns observed in natural high-strain zones that have remained unexplained by single-scale models.

Beschreibung: Vibrational densities of states and phonon dispersion relations for Mg0.875Fe0.125O ferropericlase in the high- and low-spin (HS and LS) states were calculated from first principles lattice dynamics using the internally consistent LDA+U technique. Finite-temperature thermodynamic properties were determined based on the quasi-harmonic approximation including the HS and LS mixing entropy and the magnetic entropy effects, which gave pressure and temperature variations of the low-spin fraction. Our results suggest that for thermodynamic modeling of the earth's interior, the effect of the mixed spin state cannot be ignored in the lower mantle, especially the lowermost part. The anomaly in the seismic wave velocity due to the spin crossover transition of ferropericlase, if it exists, is difficult to detect because of the wide pressure range of the transition, which is broadened by the temperature effect and the damping of the amplitude of the slow seismic wave.

Beschreibung: In the Ursa Basin, Gulf of Mexico, in situ mudstone permeability near the seafloor declines from 1.1 × 10−16 to 5.8 × 10−19 m2 over a depth of 578 m. We can reproduce this in situ permeability-porosity behavior through consolidation experiments in the laboratory. We use uniaxial constant-rate-of-strain consolidation experiments to measure permeability-porosity relationships and derive in situ permeabilities of 31 mudstone samples collected at Integrated Ocean Drilling Program (IODP) Sites U1324 and U1322. Although these mudstones have similar grain-size distributions, permeability at a given porosity varies significantly between the samples due to small variations in composition or fabric. We calculate an upscaled permeability relationship based on the observed permeability variation in the samples and determine a resultant large-scale permeability anisotropy of around 30. Based on this upscaled relationship and observations of in situ pressure, we calculate upward fluid flow rates of 0.5 mm/yr. We find that given the observed compressibility, permeability, and the geologic forcing at Ursa, overpressures are predicted as observed in the subsurface. The primary mechanism for overpressure generation at Ursa is sediment loading due to rapid burial. Low vertical permeabilities, accompanied by high sedimentation rates, can cause severe overpressure near the seafloor, which controls fluid flow and can reduce slope stability as observed in the Mississippi Canyon region. Such flow systems, especially at intermediate depths on passive margins, are important due to their control over macroscale behavior such as topographic gradient of continental slopes and submarine landslides, but have been largely understudied in the past.

Beschreibung: Available online 13 December 2012 Publication year: 2012 Source: Cell Reports Dopamine contributes to corticostriatal plasticity and motor learning. Dopamine denervation profoundly alters motor performance, as in Parkinson’s disease (PD); however, the extent to which these symptoms reflect impaired motor learning is unknown. Here, we demonstrate a D2 receptor blockade-induced aberrant learning that impedes future motor performance when dopamine signaling is restored, an effect diminished by coadministration of adenosine antagonists during blockade. We hypothesize that an inappropriate corticostriatal potentiation in striatopallidal cells of the indirect pathway underlies aberrant learning. We demonstrate synaptic potentiation in striatopallidal neurons induced by D2 blockade and diminished by application of an adenosine antagonist, consistent with behavioral observations. A neurocomputational model of the basal ganglia recapitulates the behavioral pattern and further links aberrant learning to plasticity in the indirect pathway. Thus, D2-mediated aberrant learning may contribute to motor deficits in PD, suggesting new avenues for the development of therapeutics. Graphical abstract Highlights ► Dopamine blockade induces aberrant learning that impairs future motor performance ► Aberrant learning is mediated by the D2-expressing striatopallidal pathway ► A2A antagonism protects against aberrant learning but impairs recovery ► D2 blockade induces potentiation at striatopallidal corticostriatal synapses

Beschreibung: Available online 13 December 2012 Publication year: 2012 Source: Cell Reports Elimination of aberrantly folded polypeptides from the endoplasmic reticulum (ER) by the ER - a ssociated d egradation (ERAD) system promotes cell survival under stress conditions. This quality control mechanism requires movement of misfolded proteins across the ER membrane for targeting to the cytosolic proteasome, a process facilitated by a “holdase” complex, consisting of Bag6 and the cofactors Ubl4A and Trc35. This multiprotein complex also participates in several other protein quality control processes. Here, we report SGTA as a component of the Bag6 system, which cooperates with Bag6 to channel dislocated ERAD substrates that are prone to aggregation. Using nuclear magnetic resonance spectroscopy and biochemical assays, we demonstrate that SGTA contains a noncanonical ubiquitin-like-binding domain that interacts specifically with an unconventional ubiquitin-like protein/domain in Ubl4A at least in part via electrostatics. This interaction helps recruit SGTA to Bag6, enhances substrate loading to Bag6, and thus prevents the formation of nondegradable protein aggregates in ERAD. Graphical abstract Highlights ► The two UBLs in the Bag6 complex have distinct features that specify their interactors ► The Ubl4A UBL interacts with SGTA to enhance its binding to Bag6 ► The Ubl4A UBL binds SGTA-N via an unconventional means of UBL recognition ► SGTA assists Bag6 in maintaining the solubility of ERAD substrates and promoting ERAD

Beschreibung: Available online 13 December 2012 Publication year: 2012 Source: Cell Reports The formation of the mammalian cortex requires the generation, migration, and differentiation of neurons. The vital role that the microtubule cytoskeleton plays in these cellular processes is reflected by the discovery that mutations in various tubulin isotypes cause different neurodevelopmental diseases, including lissencephaly ( TUBA1A ), polymicrogyria ( TUBA1A , TUBB2B , TUBB3 ), and an ocular motility disorder ( TUBB3 ). Here, we show that Tubb5 is expressed in neurogenic progenitors in the mouse and that its depletion in vivo perturbs the cell cycle of progenitors and alters the position of migrating neurons. We report the occurrence of three microcephalic patients with structural brain abnormalities harboring de novo mutations in TUBB5 (M299V, V353I, and E401K). These mutant proteins, which affect the chaperone-dependent assembly of tubulin heterodimers in different ways, disrupt neurogenic division and/or migration in vivo. Our results provide insight into the functional repertoire of the tubulin gene family, specifically implicating TUBB5 in embryonic neurogenesis and microcephaly. Graphical abstract Highlights ► The β-tubulin Tubb5 is highly expressed in the developing mouse and human cortex ► In vivo knockdown of Tubb5 perturbs the cell cycle and alters neuronal positioning ► Mutations in TUBB5 cause microcephaly with dysmorphic basal ganglia in humans ► TUBB5 mutations affect chaperone-mediated tubulin folding in different ways

Beschreibung: Available online 13 December 2012 Publication year: 2012 Source: Cell Reports Elimination of aberrantly folded polypeptides from the endoplasmic reticulum (ER) by the ER - a ssociated d egradation (ERAD) system promotes cell survival under stress conditions. This quality control mechanism requires movement of misfolded proteins across the ER membrane for targeting to the cytosolic proteasome, a process facilitated by a “holdase” complex, consisting of Bag6 and the cofactors Ubl4A and Trc35. This multiprotein complex also participates in several other protein quality control processes. Here, we report SGTA as a component of the Bag6 system, which cooperates with Bag6 to channel dislocated ERAD substrates that are prone to aggregation. Using nuclear magnetic resonance spectroscopy and biochemical assays, we demonstrate that SGTA contains a noncanonical ubiquitin-like-binding domain that interacts specifically with an unconventional ubiquitin-like protein/domain in Ubl4A at least in part via electrostatics. This interaction helps recruit SGTA to Bag6, enhances substrate loading to Bag6, and thus prevents the formation of nondegradable protein aggregates in ERAD. Graphical abstract Highlights ► The two UBLs in the Bag6 complex have distinct features that specify their interactors ► The Ubl4A UBL interacts with SGTA to enhance its binding to Bag6 ► The Ubl4A UBL binds SGTA-N via an unconventional means of UBL recognition ► SGTA assists Bag6 in maintaining the solubility of ERAD substrates and promoting ERAD

Beschreibung: Available online 7 December 2012 Publication year: 2012 Source: Cell Reports The efficient generation of hematopoietic stem cells from human pluripotent stem cells is dependent on the appropriate specification of the definitive hematopoietic program during differentiation. In this study, we used T lymphocyte potential to track the onset of definitive hematopoiesis from human embryonic and induced pluripotent stem cells differentiated with specific morphogens in serum- and stromal-free cultures. We show that this program develops from a progenitor population with characteristics of hemogenic endothelium, including the expression of CD34, VE-cadherin, GATA2 , LMO2 , and RUNX1 . Along with T cells, these progenitors display the capacity to generate myeloid and erythroid cells. Manipulation of Activin/Nodal signaling during early stages of differentiation revealed that development of the definitive hematopoietic progenitor population is not dependent on this pathway, distinguishing it from primitive hematopoiesis. Collectively, these findings demonstrate that it is possible to generate T lymphoid progenitors from pluripotent stem cells and that this lineage develops from a population whose emergence marks the onset of human definitive hematopoiesis. Graphical abstract Highlights ► Activin/Nodal signaling distinguishes between primitive and definitive hematopoiesis ► T cell development identifies definitive hematopoiesis in human pluripotent cultures ► Primitive hematopoiesis can be distinguished by CD41a and CD235a coexpression ► Demonstration of human T cell development from hiPSCs

Beschreibung: Available online 7 December 2012 Publication year: 2012 Source: Cell Reports DNA ligase IV (LigIV) and Artemis are central components of the nonhomologous end-joining (NHEJ) machinery that is required for V(D)J recombination and the maintenance of genomic integrity in mammalian cells. We report here crystal structures of the LigIV DNA binding domain (DBD) in both its apo form and in complex with a peptide derived from the Artemis C-terminal region. We show that LigIV interacts with Artemis through an extended hydrophobic surface. In particular, we find that the helix α2 in LigIV-DBD is longer than in other mammalian ligases and presents residues that specifically interact with the Artemis peptide, which adopts a partially helical conformation on binding. Mutations of key residues on the LigIV-DBD hydrophobic surface abolish the interaction. Together, our results provide structural insights into the specificity of the LigIV-Artemis interaction and how the enzymatic activities of the two proteins may be coordinated during NHEJ. Graphical abstract Highlights ► Crystal structure of the Ligase-IV-Artemis complex ► Ligase IV DNA binding domain contains a unique hydrophobic cavity ► The structure shows why only Ligase IV can recruit Artemis ► Insights into how activities of Ligase IV and Artemis may be coordinated in NHEJ

Beschreibung: 29 November 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 5 A key feature of RNA polymerase II (Pol II) preinitiation complexes (PICs) is their ability to coordinate transcription initiation with chromatin modification and remodeling. To understand how this coordination is achieved, we employed extensive proteomic and mechanistic analyses to study the composition and assembly of PICs in HeLa cell and mouse embryonic stem cell (ESC) nuclear extracts. Strikingly, most of the machinery that is necessary for transcription initiation on chromatin is part of the PIC. The PIC is nearly identical between ESCs and HeLa cells and contains two major coactivator complexes: Mediator and SAGA. Genome-wide analysis of Mediator reveals that it has a close correlation with Pol II, TATA-binding protein, and messenger RNA levels and thus may play a major role in PIC assembly. Moreover, Mediator coordinates assembly of the Pol II initiation factors and chromatin machinery into a PIC in vitro, whereas SAGA acts after PIC assembly to allow transcription on chromatin. Graphical abstract Highlights ► The composition of mammalian PICs was determined by proteomic analysis ► A wide range of chromatin-modifying and -remodeling factors are recruited to PICs ► Mediator coordinates binding of Pol II initiation, elongation, and chromatin machineries ► SAGA acts after PIC assembly to make chromatin templates transcriptionally competent

Beschreibung: 29 November 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 5 In meiosis, two specialized cell divisions allow the separation of paired chromosomes first, then of sister chromatids. Separase removes the cohesin complex holding sister chromatids together in a stepwise manner from chromosome arms in meiosis I, then from the centromere region in meiosis II. Using mouse oocytes, our study reveals that cyclin A2 promotes entry into meiosis, as well as an additional unexpected role; namely, its requirement for separase-dependent sister chromatid separation in meiosis II. Untimely cyclin A2-associated kinase activity in meiosis I leads to precocious sister separation, whereas inhibition of cyclin A2 in meiosis II prevents it. Accordingly, endogenous cyclin A is localized to kinetochores throughout meiosis II, but not in anaphase I. Additionally, we found that cyclin B1, but not cyclin A2, inhibits separase in meiosis I. These findings indicate that separase-dependent cohesin removal is differentially regulated by cyclin B1 and A2 in mammalian meiosis. Graphical abstract Highlights ► Cyclin A2 is required for meiotic entry and sister chromatid separation in meiosis II ► Constitutive cyclin A2 activity in meiosis I leads to precocious sister separation ► Only cyclin B1, and not cyclin A2, can inhibit separase in mouse oocyte meiosis I ► Endogenous cyclin A is localized to centromeres throughout the second meiotic division

Beschreibung: Available online 6 December 2012 Publication year: 2012 Source: Cell Reports Pathogens that reside in the phagosomes of infected cells persist despite the presence of potent T cell responses. We addressed the mechanism of immune evasion by using a mouse model of Salmonella typhimurium (ST). Recombinants of ST were generated that translocated antigen to the cytosol or phagosomes of infected cells. We find that the kinetics of antigen presentation and CD8 + T cell priming is accelerated by cytosolic antigen delivery, although the magnitude of CD8 + T cell response is not influenced by antigenic location. More importantly, only those targets that readily display antigen on the cell surface, owing to antigenic translocation to the cytosol, are recognized and killed by CD8 + T cells. Thus, vaccination approaches developed to control phagosomal pathogens should incorporate methods for modulating antigen presentation such that infected target cells can be readily recognized by CD8 + T cells. Graphical abstract Highlights ► Intracellular location of antigen governs the duration of infection ► Forcing the recognition of infected cells by CD8 T cells controls infection ► CD8 T cells can convert a chronic infection to an acute infection ► Suppression is not the cause, but the end result, of a chronic infection

Beschreibung: Available online 13 December 2012 Publication year: 2012 Source: Cell Reports Trafficking of proteins specifically to the axonal or somatodendritic membrane allows neurons to establish and maintain polarized compartments with distinct morphology and function. Diverse evidence suggests that an actin-dependent vesicle filter within the axon initial segment (AIS) plays a critical role in polarized trafficking; however, no distinctive actin-based structures capable of comprising such a filter have been found within the AIS. Here, using correlative light and scanning electron microscopy, we visualized networks of actin filaments several microns wide within the AIS of cortical neurons in culture. Individual filaments within these patches are predominantly oriented with their plus ends facing toward the cell body, consistent with models of filter selectivity. Vesicles carrying dendritic proteins are much more likely to stop in regions occupied by the actin patches than in other regions, indicating that the patches likely prevent movement of dendritic proteins to the axon and thereby act as a vesicle filter. Graphical abstract Highlights ► 2 μm diameter actin networks are visualized by SEM and light microscopy in the AIS ► Actin filaments within networks are oriented with their plus ends facing proximally ► Actin network structure suggests a mechanism for selectively halting vesicles ► Vesicles carrying dendritic proteins halt and reverse in actin networks

Beschreibung: Available online 13 December 2012 Publication year: 2012 Source: Cell Reports Dopamine contributes to corticostriatal plasticity and motor learning. Dopamine denervation profoundly alters motor performance, as in Parkinson’s disease (PD); however, the extent to which these symptoms reflect impaired motor learning is unknown. Here, we demonstrate a D2 receptor blockade-induced aberrant learning that impedes future motor performance when dopamine signaling is restored, an effect diminished by coadministration of adenosine antagonists during blockade. We hypothesize that an inappropriate corticostriatal potentiation in striatopallidal cells of the indirect pathway underlies aberrant learning. We demonstrate synaptic potentiation in striatopallidal neurons induced by D2 blockade and diminished by application of an adenosine antagonist, consistent with behavioral observations. A neurocomputational model of the basal ganglia recapitulates the behavioral pattern and further links aberrant learning to plasticity in the indirect pathway. Thus, D2-mediated aberrant learning may contribute to motor deficits in PD, suggesting new avenues for the development of therapeutics. Graphical abstract Highlights ► Dopamine blockade induces aberrant learning that impairs future motor performance ► Aberrant learning is mediated by the D2-expressing striatopallidal pathway ► A2A antagonism protects against aberrant learning but impairs recovery ► D2 blockade induces potentiation at striatopallidal corticostriatal synapses

Beschreibung: Available online 13 December 2012 Publication year: 2012 Source: Cell Reports Sir2, an evolutionarily conserved NAD + -dependent deacetylase, has been implicated as a key factor in mediating organismal life span. However, recent contradictory findings have brought into question the role of Sir2 and its orthologs in regulating organismal longevity. In this study, we report that Drosophila Sir2 (dSir2) in the adult fat body regulates longevity in a diet-dependent manner. We used inducible Gal4 drivers to knock down and overexpress dSir2 in a tissue-specific manner. A diet-dependent life span phenotype of dSir2 perturbations (both knockdown and overexpression) in the fat body, but not muscles, negates the effects of background genetic mutations. In addition to providing clarity to the field, our study contrasts the ability of dSir2 in two metabolic tissues to affect longevity. We also show that dSir2 knockdown abrogates fat-body dFOXO-dependent life span extension. This report highlights the importance of the interplay between genetic factors and dietary inputs in determining organismal life spans. Graphical abstract Highlights ► The dSir2-dependent longevity phenotype is diet specific ► DR-dependent effects are associated with increased dSir2 expression and NAD + levels ► dSir2 in the fat body, but not in muscles, regulates longevity ► dSir2-dFOXO interaction in the fat body affects life-span extension

Beschreibung: Available online 6 December 2012 Publication year: 2012 Source: Cell Reports Deubiquitinating enzymes (DUBs) constitute a large family of cysteine proteases that have a broad impact on numerous biological and pathological processes, including the regulation of genomic stability. DUBs are often assembled onto multiprotein complexes to assist in their localization and substrate selection, yet it remains unclear how the enzymatic activity of DUBs is modulated by intracellular signals. Herein, we show that bursts of reactive oxygen species (ROS) reversibly inactivate DUBs through the oxidation of the catalytic cysteine residue. Importantly, USP1, a key regulator of genomic stability, is reversibly inactivated upon oxidative stress. This, in part, explains the rapid nature of PCNA monoubiquitination-dependent DNA damage tolerance in response to oxidative DNA damage in replicating cells. We propose that DUBs of the cysteine protease family act as ROS sensors in human cells and that ROS-mediated DUB inactivation is a critical mechanism for fine-tuning stress-activated signaling pathways. Graphical abstract Highlights ► USP1 is inactivated upon oxidative stress to fine-tune PCNA monoubiquitination ► The Cys protease family of DUBs are reversibly inactivated by ROS ► The AMSH metalloprotease DUB is refractory to ROS-mediated catalytic inactivation

Beschreibung: Available online 13 December 2012 Publication year: 2012 Source: Cell Reports Genome information, which is three-dimensionally organized within cells as chromatin, is searched and read by various proteins for diverse cell functions. Although how the protein factors find their targets remains unclear, the dynamic and flexible nature of chromatin is likely crucial. Using a combined approach of fluorescence correlation spectroscopy, single-nucleosome imaging, and Monte Carlo computer simulations, we demonstrate local chromatin dynamics in living mammalian cells. We show that similar to interphase chromatin, dense mitotic chromosomes also have considerable chromatin accessibility. For both interphase and mitotic chromatin, we observed local fluctuation of individual nucleosomes (∼50 nm movement/30 ms), which is caused by confined Brownian motion. Inhibition of these local dynamics by crosslinking impaired accessibility in the dense chromatin regions. Our findings show that local nucleosome dynamics drive chromatin accessibility. We propose that this local nucleosome fluctuation is the basis for scanning genome information. Graphical abstract Highlights ► Dense chromatin regions have considerable chromatin accessibility ► Observed local fluctuation of individual nucleosomes in interphase and mitotic chromatin ► Inhibition of nucleoome fluctuation impaired the chromatin accessibility ► Local fluctuation of nucleosomes is the basis for scanning genome information

Beschreibung: 29 November 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 5 Expansions of simple DNA repeats cause numerous hereditary diseases in humans. We analyzed the role of DNA polymerases in the instability of Friedreich’s ataxia (GAA) n repeats in a yeast experimental system. The elementary step of expansion corresponded to ∼160 bp in the wild-type strain, matching the size of Okazaki fragments in yeast. This step increased when DNA polymerase α was mutated, suggesting a link between the scale of expansions and Okazaki fragment size. Expandable repeats strongly elevated the rate of mutations at substantial distances around them, a phenomenon we call repeat-induced mutagenesis (RIM). Notably, defects in the replicative DNA polymerases δ and ε strongly increased rates for both repeat expansions and RIM. The increases in repeat-mediated instability observed in DNA polymerase δ mutants depended on translesion DNA polymerases. We conclude that repeat expansions and RIM are two sides of the same replicative mechanism. Graphical abstract Highlights ► Elementary step of repeat expansion corresponds to size of an Okazaki fragment ► Mutated DNA polymerase α leads to an increase in expansion step ► Mutated DNA polymerases δ and ε lead to elevated expansion rates ► Repeats induce mutagenesis, which is further elevated in polymerase mutants

Beschreibung: 29 November 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 5 Plasma sphingosine-1-phosphate (S1P) regulates vascular permeability, and plasma and lymph S1P guide lymphocyte egress from lymphoid organs. S1P is made intracellularly, and little is known about how S1P is delivered into circulatory fluids. Here, we find that mice without the major facilitator superfamily transporter Spns2 have a profound reduction in lymph S1P, but only a minor decrease in plasma S1P. Spns2-deficient mice have a redistribution of lymphocytes from the spleen to lymph nodes and a loss of circulating lymphocytes, consistent with normal egress from the spleen directed by plasma S1P and blocked egress from lymph nodes directed by lymph S1P. Spns2 is needed in endothelial cells to supply lymph S1P and support lymphocyte circulation. As a differential requirement for lymph and blood S1P, Spns2 may be an attractive target for immune suppressive drugs. Graphical abstract Highlights ► The transporter Spns2 is required to supply lymph, but not plasma, S1P ► Spns2-deficient mice have disrupted peripheral lymphocyte circulation ► Spns2 is required in endothelial cells to secrete lymph S1P and support trafficking

Beschreibung: 29 November 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 5 Elongin A increases the rate of RNA polymerase II (pol II) transcript elongation by suppressing transient pausing by the enzyme. Elongin A also acts as a component of a cullin-RING ligase that can target stalled pol II for ubiquitylation and proteasome-dependent degradation. It is not known whether these activities of Elongin A are functionally interdependent in vivo. Here, we demonstrate that Elongin A-deficient (Elongin A −/− ) embryos exhibit abnormalities in the formation of both cranial and spinal nerves and that Elongin A −/− embryonic stem cells (ESCs) show a markedly decreased capacity to differentiate into neurons. Moreover, we identify Elongin A mutations that selectively inactivate one or the other of the aforementioned activities and show that mutants that retain the elongation stimulatory, but not pol II ubiquitylation, activity of Elongin A rescue neuronal differentiation and support retinoic acid-induced upregulation of a subset of neurogenesis-related genes in Elongin A −/− ESCs. Graphical abstract Highlights ► RA-induced neuronal differentiation is markedly impaired in Elongin A −/− ESCs ► Formation of cranial ganglia and DRG is severely impaired in Elongin A −/− embryos ► Mutations that differentially affect the two activities of Elongin A were identified ► Elongin A’s elongation stimulatory activity is required for a subset of RA-induced genes

Beschreibung: 29 November 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 5 The large Mediator (L-Mediator) is a general coactivator of RNA polymerase II transcription and is formed by the reversible association of the small Mediator (S-Mediator) and the kinase-module-harboring Cdk8. It is not known how the kinase module association/dissociation is regulated. We describe the fission yeast Cdk11-L-type cyclin pombe (Lcp1) complex and show that its inactivation alters the global expression profile in a manner very similar to that of mutations of the kinase module. Cdk11 is broadly distributed onto chromatin and phosphorylates the Med27 and Med4 Mediator subunits on conserved residues. The association of the kinase module and the S-Mediator is strongly decreased by the inactivation of either Cdk11 or the mutation of its target residues on the Mediator. These results show that Cdk11-Lcp1 regulates the association of the kinase module and the S-Mediator to form the L-Mediator complex. Graphical abstract Highlights ► A complex related to metazoan Cdk11-CyclinL is conserved in fission yeast ► Loss of either Cdk8 or Cdk11 similarly affects global expression profile ► Cdk11 phosphorylates two Mediator subunits ► Association of the Cdk8 module with the S-Mediator requires phosphorylation by Cdk11

Beschreibung: 29 November 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 5 Age-related macular degeneration (AMD) is the leading cause of irreversible blindness in the elderly population worldwide. Although recent studies have demonstrated strong genetic associations between AMD and SNPs in a number of genes, other modes of regulation are also likely to play a role in the etiology of this disease. We identified a significantly decreased level of methylation on the IL17RC promoter in AMD patients. Furthermore, we showed that hypomethylation of the IL17RC promoter in AMD patients led to an elevated expression of its protein and messenger RNA in peripheral blood as well as in the affected retina and choroid, suggesting that the DNA methylation pattern and expression of IL17RC may potentially serve as a biomarker for the diagnosis of AMD and likely plays a role in disease pathogenesis. Graphical abstract Highlights ► ∼1.5% of CpG sites are differentially methylated between twins with discordant AMD ► Hypomethylated IL17RC promoter and elevated IL17RC expression are associated with AMD ► IL-17-mediated inflammatory responses contribute to AMD pathogenesis

Beschreibung: 29 November 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 5 RIG-I and MDA5 are cytosolic RNA sensors that play a critical role in innate antiviral responses. Major advances have been made in identifying RIG-I ligands, but our knowledge of the ligands for MDA5 remains restricted to data from transfection experiments mostly using poly(I:C), a synthetic dsRNA mimic. Here, we dissected the IFN-α/β-stimulatory activity of different viral RNA species produced during picornavirus infection, both by RNA transfection and in infected cells in which specific steps of viral RNA replication were inhibited. Our results show that the incoming genomic plus-strand RNA does not activate MDA5, but minus-strand RNA synthesis and production of the 7.5 kbp replicative form trigger a strong IFN-α/β response. IFN-α/β production does not rely on plus-strand RNA synthesis and thus generation of the partially double-stranded replicative intermediate. This study reports MDA5 activation by a natural RNA ligand under physiological conditions. Graphical abstract Highlights ► Viral ssRNA, with or without the 5′ VPg peptide, does not induce IFN-α/β ► Synthesis of minus-strand—but not plus-strand—RNA is crucial for MDA5 activation ► The picornavirus RF activates MDA5 both in vitro and in vivo ► MDA5 activation by viral replicative form is independent of the terminal groups of this RNA

Beschreibung: 29 November 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 5 In mammals, each olfactory bulb (OB) contains a pair of mirror-symmetric glomerular maps organized to reflect odorant receptor identity. The functional implication of maintaining these symmetric medial-lateral maps within each OB remains unclear. Here, using in vivo multielectrode recordings to simultaneously detect odorant-induced activity across the entire OB, we reveal a timing difference in the odorant-evoked onset latencies between the medial and lateral halves. Interestingly, the latencies in the medial and lateral OB decreased at different rates as odorant concentration increased, causing the timing difference between them to also diminish. As a result, output neurons in the medial and lateral OB fired with greater synchrony at higher odorant concentrations. Thus, we propose that temporal differences in activity between the medial and lateral OB can dynamically code odorant concentration, which is subsequently decoded in the olfactory cortex through the integration of synchronous action potentials. Graphical abstract Highlights ► Odorants differentially activate the medial and lateral olfactory bulb (OB) ► An ∼60 ms time window is defined between medial and lateral mitral cell output ► The medial-lateral time window is dynamically modulated by odorant concentrations ► High odorant concentrations synchronize medial-lateral OB output

Beschreibung: 29 November 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 5 MicroRNAs (miRNAs) are essential regulators of development, physiology, and evolution, and their biogenesis is strictly controlled at multiple levels. Regulatory proteins, such as KSRP, modulate rates and timing of enzymatic reactions responsible for maturation of select miRNAs from their primary transcripts in response to specific stimuli. Here, we show that KSRP silencing in mesenchymal C2C12 cells produces a change in the transcriptome largely overlapping that induced by bone morphogenetic protein 2 (BMP2) signaling activation. This induces osteoblastic differentiation while preventing myogenic differentiation. KSRP silencing- and BMP2-dependent myogenic miRNA (myomiR) maturation blockade is required for osteoblastic differentiation of C2C12 cells. Our results demonstrate that phosphorylated R-SMAD proteins, the transducers of BMP2 signal, associate with phosphorylated KSRP and block its interaction with primary myomiRs. This abrogates KSRP-dependent myomiR maturation, with SMAD4, SMAD5, and SMAD9 silencing being able to rescue KSRP function. Thus, SMAD-induced blockade of KSRP-dependent myomiR maturation is critical for orienting C2C12 cell differentiation toward osteoblastic lineage. Graphical abstract Highlights ► KSRP silencing and BMP2 signaling activation block myomiR maturation ► myomiR maturation blockade is required for osteoblastic differentiation of C2C12 cells ► SMAD proteins interact with KSRP and block its ability to promote myomiR maturation

Beschreibung: 29 November 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 5 Senescence is a cellular response preventing tumorigenesis. The Ras oncogene is frequently activated or mutated in human cancers, but Ras activation is insufficient to transform primary cells. In a search for cooperating oncogenes, we identify the lysine demethylase JMJD2A/KDM4A. We show that JMJD2A functions as a negative regulator of Ras-induced senescence and collaborates with oncogenic Ras to promote cellular transformation by negatively regulating the p53 pathway. We find CHD5 , a known tumor suppressor regulating p53 activity, as a target of JMJD2A. The expression of JMJD2A inhibits Ras-mediated CHD5 induction leading to a reduced activity of the p53 pathway. In addition, we show that JMJD2A is overexpressed in mouse and human lung cancers. Depletion of JMJD2A in the human lung cancer cell line A549 bearing an activated K-Ras allele triggers senescence. We propose that JMJD2A is an oncogene that represents a target for Ras-expressing tumors. Graphical abstract Highlights ► JMJD2A contributes to the bypass of Ras-induced senescence ► Genome-wide search for gene promoters regulated by JMJD2A identifies CHD5 ► JMJD2A cooperates with Ras to transform primary cells ► JMJD2A depletion in human lung cancer cells leads to massive senescence

Beschreibung: 29 November 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 5 Mitochondria-originating reactive oxygen species (ROS) control T cell receptor (TCR)-induced gene expression. Here, we show that TCR-triggered activation of ADP-dependent glucokinase (ADPGK), an alternative, glycolytic enzyme typical for Archaea , mediates generation of the oxidative signal. We also show that ADPGK is localized in the endoplasmic reticulum and suggest that its active site protrudes toward the cytosol. The ADPGK-driven increase in glycolytic metabolism coincides with TCR-induced glucose uptake, downregulation of mitochondrial respiration, and deviation of glycolysis toward mitochondrial glycerol-3-phosphate dehydrogenase (GPD) shuttle; i.e., a metabolic shift to aerobic glycolysis similar to the Warburg effect. The activation of respiratory-chain-associated GPD2 results in hyperreduction of ubiquinone and ROS release from mitochondria. In parallel, mitochondrial bioenergetics and ultrastructure are altered. Downregulation of ADPGK or GPD2 abundance inhibits oxidative signal generation and induction of NF-κB-dependent gene expression, whereas overexpression of ADPGK potentiates them. Graphical abstract Highlights ► TCR triggering activates ADPGK, an ER-localized glycolytic enzyme ► TCR induction shifts metabolism toward glycolysis and activates mitochondrial GPD2 ► Activation of GPD2 leads to ubiquinol accumulation and ROS release from mitochondria ► ADPGK- and GPD2-dependent oxidative signal drives NF-κB-regulated gene expression

Beschreibung: 29 November 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 5 Alzheimer's disease (AD) is the most common cause of dementia among older people. There are no effective medications currently available to prevent and treat AD and halt disease progression. Monoacylglycerol lipase (MAGL) is the primary enzyme metabolizing the endocannabinoid 2-arachidonoylglycerol in the brain. We show here that inactivation of MAGL robustly suppressed production and accumulation of β-amyloid (Aβ) associated with reduced expression of β-site amyloid precursor protein cleaving enzyme 1 (BACE1) in a mouse model of AD. MAGL inhibition also prevented neuroinflammation, decreased neurodegeneration, maintained integrity of hippocampal synaptic structure and function, and improved long-term synaptic plasticity, spatial learning, and memory in AD animals. Although the molecular mechanisms underlying the beneficial effects produced by MAGL inhibition remain to be determined, our results suggest that MAGL, which regulates endocannabinoid and prostaglandin signaling, contributes to pathogenesis and neuropathology of AD, and thus is a promising therapeutic target for the prevention and treatment of AD. Graphical abstract Highlights ► Inactivation of MAGL reduces Aβ plaques and BACE1 expression in AD mice ► MAGL inhibition decreases neuroinflammation and neurodegeneration ► MAGL inhibition maintains integrity of hippocampal synaptic structure and function ► MAGL inhibition improves synaptic plasticity and learning and memory in AD mice

Beschreibung: 29 November 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 5 Antagonistic pleiotropy (AP), or genetic tradeoff, is an important concept that is frequently invoked in theories of aging, cancer, genetic disease, and other common phenomena. However, the prevalence of AP, which genes are subject to AP, and to what extent and how AP may be resolved remain unclear. By measuring the fitness difference between the wild-type and null alleles of ∼5,000 nonessential genes in yeast, we found that in any given environment, yeast expresses hundreds of genes that harm rather than benefit the organism, demonstrating widespread AP. Nonetheless, under sufficient selection, AP is often resolvable through regulatory evolution, primarily by trans -acting changes, although in one case we also detected a cis -acting change and localized its causal mutation. However, AP is resolved more slowly in smaller populations, predicting more unresolved AP in multicellular organisms than in yeast. These findings provide an empirical foundation for AP-dependent theories and have broad biomedical and evolutionary implications. Graphical abstract Highlights ► Under any conditions, yeast expresses many genes that are harmful to the cell ► Such problems can often be resolved by regulatory evolution under selection ► Such regulatory evolution tends to occur via trans -acting genetic changes ► Antagonistic pleiotropy is predicted to be more abundant in multicellular organisms

Beschreibung: 29 November 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 5 After implantation, pluripotent epiblasts are converted to embryonic ectoderm through cell–cell interactions that significantly change the transcriptional and epigenetic networks. An entrée to understanding this vital developmental transition is the t w5 mutation of the mouse t complex. This mutation produces highly specific defects in the embryonic ectoderm before gastrulation, leading to death of the embryonic ectoderm. Using a positional cloning approach, we have now identified the mutated gene, completing a decades-long search. The gene, vacuolar protein sorting 52 ( Vps52 ), is a mouse homolog of yeast VPS52 that is involved in the retrograde trafficking of endosomes. Our data suggest that Vps52 acts in extraembryonic tissues to support the growth and differentiation of embryonic ectoderm via cell–cell interactions. It is also required in the formation of embryonic structures at a later stage of development, revealing hitherto unknown functions of Vps52 in the development of a multicellular organism. Graphical abstract Highlights ► Vps52 , part of retrograde transport to the Golgi, causes the t w5 lethal phenotype ► Vps52 acts in extraembryonic tissues to dictate embryonic ectoderm differentiation ► VPS52’s essential function in the embryo involves cell–cell interactions ► VPS52 also has an indispensable function later in development

Beschreibung: 29 November 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 5 Human pluripotent stem cells (hPSCs), including embryonic stem cells and induced pluripotent stem cells, are potentially useful in regenerative therapies for heart disease. For medical applications, clinical-grade cardiac cells must be produced from hPSCs in a defined, cost-effective manner. Cell-based screening led to the discovery of KY02111, a small molecule that promotes differentiation of hPSCs to cardiomyocytes. Although the direct target of KY02111 remains unknown, results of the present study suggest that KY02111 promotes differentiation by inhibiting WNT signaling in hPSCs but in a manner that is distinct from that of previously studied WNT inhibitors. Combined use of KY02111 and WNT signaling modulators produced robust cardiac differentiation of hPSCs in a xeno-free, defined medium, devoid of serum and any kind of recombinant cytokines and hormones, such as BMP4, Activin A, or insulin. The methodology has potential as a means for the practical production of human cardiomyocytes for regeneration therapies. Graphical abstract Highlights ► KY02111 promotes the differentiation of hPSCs to functional cardiomyocytes ► KY02111 acts downstream of APC and GSK3β to inhibit WNT signaling ► KY02111 and WNT inhibitors cooperatively enhance hPSC cardiomyogenesis ► KY02111and WNT modulators permit cytokine and xeno-free hPSC cardiomyogenesis

Beschreibung: 29 November 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 5 Growing evidence suggests that aggregation-prone proteins are both harmful and functional for a cell. How do cellular systems balance the detrimental and beneficial effect of protein aggregation? We reveal that aggregation-prone proteins are subject to differential transcriptional, translational, and degradation control compared to nonaggregation-prone proteins, which leads to their decreased synthesis, low abundance, and high turnover. Genetic modulators that enhance the aggregation phenotype are enriched in genes that influence expression homeostasis. Moreover, genes encoding aggregation-prone proteins are more likely to be harmful when overexpressed. The trends are evolutionarily conserved and suggest a strategy whereby cellular mechanisms specifically modulate the availability of aggregation-prone proteins to (1) keep concentrations below the critical ones required for aggregation and (2) shift the equilibrium between the monomeric and oligomeric/aggregate form, as explained by Le Chatelier’s principle. This strategy may prevent formation of undesirable aggregates and keep functional assemblies/aggregates under control. Graphical abstract Highlights ► mRNA encoding aggregation-prone proteins is complex, suggesting greater translational regulation ► Aggregation-prone proteins are present in low abundance and for short periods of time ► Tight control is evolutionarily conserved and provides robustness against aggregation ► Aggregation-prone proteins are subject to tight regulation

Beschreibung: 29 November 2012 Publication year: 2012 Source: Cell Reports, Volume 2, Issue 5 Variability in signaling pathway activation between neighboring epithelial cells can arise from local differences in the microenvironment, noisy gene expression, or acquired genetic changes. To investigate the consequences of this cell-to-cell variability in signaling pathway activation on coordinated multicellular processes such as morphogenesis, we use DNA-programmed assembly to construct three-dimensional MCF10A microtissues that are mosaic for low-level expression of activated H-Ras. We find two emergent behaviors in mosaic microtissues: cells with activated H-Ras are basally extruded or lead motile multicellular protrusions that direct the collective motility of their wild-type neighbors. Remarkably, these behaviors are not observed in homogeneous microtissues in which all cells express the activated Ras protein, indicating that heterogeneity in Ras activity, rather than the total amount of Ras activity, is critical for these processes. Our results directly demonstrate that cell-to-cell variability in pathway activation within local populations of epithelial cells can drive emergent behaviors during epithelial morphogenesis. Graphical abstract Highlights ► Cell-to-cell variability in Ras activity triggers emergent behaviors ► Homogeneous and low-level Ras activation does not disrupt early morphogenesis ► Programmed assembly precisely controls the composition of mosaic epithelial tissues

Beschreibung: Explosive activity at Stromboli volcano is analyzed using a high-frame rate (50 Hz) thermal camera and differential pressure transducers. We develop a thermal image-based decomposition method to derive vertical and horizontal exit velocities of the explosive cloud. Peak vertical velocity ranges between 23 and 203 m/s, slightly higher than previous estimates and rapidly decreasing to a constant value of 30–50 m/s within the first ∼0.1 s. Plume velocities are consistent with an elongated cloud expanding much faster vertically than horizontally and indicating the interaction with the conduit wall. Considering a vent radius of ∼2 m we estimate a volumetric flux of 200–600 m3/s, which converts to total volumes of gas-particles of 103–104 m3 for a single eruption. These volumes are proportional to the thermal energy recorded by the camera, providing a means to convert thermal radiance to volumes. Comparing the thermal onset of the explosions with the arrival time of the acoustic pressure, we demonstrate that infrasound is propagating 0.14−1.7 s ahead of the explosive front. The time difference between thermal and acoustic onsets constrains the infrasonic source within the conduit at 15–35 m below the crater rim. Peak amplitudes of acoustic pressure show a power law relationship (p ∼ U2) with the exit vertical velocities consistent with the energy balance of a two-phase flow rapidly accelerated in the conduit by gas decompression. Our results support monopole isotropic acoustic radiation of a source embedded within the conduit walls and indicate that explosive dynamics undergo strong accelerations of 103–104 m/s2.

Beschreibung: We present a high resolution 3 dimensional (3D) P wave velocity model for Tenerife Island, Canaries, covering the top of Teide volcano (3,718 m a.s.l.) down to around 8 km below sea level (b.s.l). The tomographic inversion is based on a large data set of travel times obtained from a 3D active seismic experiment using offshore shots (air guns) recorded at more than 100 onshore seismic stations. The obtained seismic velocity structure is strongly heterogeneous with significant (up to 40%) lateral variations. The main volcanic structure of the Las Cañadas-Teide-Pico Viejo Complex (CTPVC) is characterized by a high P wave velocity body, similar to many other stratovolcanoes. The presence of different high P wave velocity regions inside the CTPVC may be related to the geological and volcanological evolution of the system. The presence of high P wave velocities at the center of the island is interpreted as evidence for a single central volcanic source for the formation of Tenerife. Furthermore, reduced P wave velocities are found in a small confined region in CTPVC and are more likely related to hydrothermal alteration, as indicated by the existence of fumaroles, than to the presence of a magma chamber beneath the system. In the external regions, surrounding CTPVC a few lower P wave velocity regions can be interpreted as fractured zones, hydrothermal alterations, porous materials and thick volcaniclastic deposits.

Beschreibung: Eastern Indonesia and the southern Philippines comprise a huge and seismically highly active region that has received less than the deserved attention in tsunami research compared with the surrounding areas exposed to the major subduction zones. In an effort to redress the balance the tsunami hazard in this region is studied by establishing a tsunami event database which, in combination with seismological and tectonic information from the region, has allowed us to define and justify a number of ‘credible worst-case’ tsunami scenarios. These scenarios have been used in numerical simulations of tsunami generation and propagation to study maximum water level along potentially affected shorelines. The scenarios have in turn been combined to provide regional tsunami hazard maps. In many cases the simulations indicate that the maximum water level may exceed 10 m locally and even reach above 20 m in the vicinity of the source, which is of the same order as what is forecasted along the Sumatra and Java trenches for comparable return periods. For sections of coastlines close to a source, a tsunami may strike only a few minutes after it is generated, providing little time for warning. Moreover, several of the affected areas are highly populated and are therefore also high risk areas. The combination of high maximum water levels, short warning times, dense populations, and relatively short return periods suggests strongly that the tsunami hazard and risk in these regions are alarmingly high.

Beschreibung: We determine the geographical boundary and average shear velocity structure of the Pacific Anomaly at the base of the mantle based on travel time analysis of ScSH-SH and ScS2 (ScSScS)-SS phases and waveform modeling results. We further constrain the detailed geometry of the northern Anomaly around (20°N, −170°E) and its transition to the surrounding high velocity region along three perpendicular cross sections on the basis of forward waveform modeling of the observed direct S and ScS phases. The observed differential travel-time residuals and waveform modeling results allow the whole geographic boundary of the Anomaly to be delineated and the area of the base of the Anomaly is estimated to be 1.9 × 107 km2. The maximum shear velocity perturbation inside the Anomaly reaches −5% in the lowermost 500 km of the mantle. Waveform analysis suggests that the northern Anomaly reaches 450 km above the CMB with both steeply and shallowly dipping edges and its basal layer extends beneath the surrounding mantle with the degree of extension changing rapidly across a small distance. The inferred characteristics of the Anomaly support the previous suggestion that the Pacific Anomaly represents a chemical anomaly. However, unlike the inferred features of the African Anomaly pointing to an ancient compositionally distinct and geologically stable anomaly, the existence of several separated piles extending into the mid-lower mantle, the complex morphology of the piles with both steeply and shallowly dipping edges and the presence of many ultra-low velocity zones at its base suggest that the Pacific Anomaly likely possesses varying intrinsic compositions and exhibits complex interaction with the surrounding mantle.

Beschreibung: Publication year: 2012 Source: Cell Reports Vadim Shchepachev, Harry Wischnewski, Edoardo Missiaglia, Charlotte Soneson, Claus M. Azzalin Clericuzio-type poikiloderma with neutropenia (PN) is a rare genodermatosis associated with mutations in the C16orf57 gene, which codes for the uncharacterized protein hMpn1. We show here that, in both fission yeasts and humans, Mpn1 processes the spliceosomal U6 small nuclear RNA (snRNA) posttranscriptionally. In Mpn1-deficient cells, U6 molecules carry 3′ end polyuridine tails that are longer than those in normal cells and lack a terminal 2′,3′ cyclic phosphate group. In mpn1 Δ yeast cells, U6 snRNA and U4/U6 di-small nuclear RNA protein complex levels are diminished, leading to precursor messenger RNA splicing defects, which are reverted by expression of either yeast or human Mpn1 and by overexpression of U6. Recombinant hMpn1 is a 3′-to-5′ RNA exonuclease that removes uridines from U6 3′ ends, generating terminal 2′,3′ cyclic phosphates in vitro. Finally, U6 degradation rates increase in mpn1 Δ yeasts and in lymphoblasts established from individuals affected by PN. Our data indicate that Mpn1 promotes U6 stability through 3′ end posttranscriptional processing and implicate altered U6 metabolism as a potential mechanism for PN pathogenesis. Graphical abstract Graphical Abstract Highlights ► Yeast and human cells deficient for Mpn1 fail to process U6 snRNA oligo(U) 3′ ends ► Cellular U6 levels and pre-mRNA splicing are compromised in mpn1 Δ yeasts ► Mpn1 is a 3′-to-5′ RNA exonuclease ► U6 degradation rates are accelerated in yeast and human cells deficient for Mpn1

Beschreibung: Publication year: 2012 Source: Cell Reports Agathe Valluet, Sabine Druillennec, Céline Barbotin, Coralie Dorard, Anne H. Monsoro-Burq, Magalie Larcher, Celio Pouponnot, Manuela Baccarini, Lionel Larue, Alain Eychène B-Raf and C-Raf kinases have emerged as critical players in melanoma. However, little is known about their role during development and homeostasis of the melanocyte lineage. Here, we report that knockout of B-raf and C-raf genes in this lineage results in normal pigmentation at birth with no defect in migration, proliferation, or differentiation of melanoblasts in mouse hair follicles. In contrast, the double raf knockout mice displayed hair graying resulting from a defect in cell-cycle entry of melanocyte stem cells (MSCs) and their subsequent depletion in the hair follicle bulge. Therefore, Raf signaling is dispensable for early melanocyte lineage development, but necessary for MSC maintenance. Graphical abstract Graphical Abstract Highlights ► Direct in vivo evidence of the involvement of Raf proteins in stemness ► B-Raf and C-Raf are required for melanocyte stem cell self-maintenance ► The Raf/MEK/ERK pathway is not essential for melanocyte lineage development ► SCF/Kit and Raf/ERK signaling pathways are uncoupled in the melanocyte lineage

Beschreibung: Publication year: 2012 Source: Cell Reports, Volume 2, Issue 3 Simone Prömel, Marie Frickenhaus, Samantha Hughes, Lamia Mestek, David Staunton, Alison Woollard, Ioannis Vakonakis, Torsten Schöneberg, Ralf Schnabel, Andreas P. Russ, Tobias Langenhan

Beschreibung: Using seismic profiles of triplicated waveforms, we show significant lateral variations in the SH velocity (Vs) structure of the transition zone (TZ) beneath Korea and adjacent regions. Beneath Sakhalin, we detected a high Vs anomaly (∼2%) limited to middle regions of the TZ (mid-TZ), and a large Vs jump across the 660-km discontinuity. A similar jump in Vs also occurs beneath the northern portion of the North China Craton (NCC). Beneath Korea, a high Vs anomaly (∼2%) in the lower TZ is inferred, accompanied by a relatively small Vs jump across the 660-km discontinuity, which is depressed by about ∼15–20 km. The deep structure under the eastern part of northeast China (NEC) also includes a slight Vs anomaly (∼1%) in the lower TZ but does not exhibit significant depression of the 660-km discontinuity. Compared with previous study, our observations reveal strong regional variations of the TZ structure on a relatively short scale. These variations most likely reflect the geometrical distribution of the subducting northwest Pacific plate. Our results suggest that the subducting slab dips across the mid-TZ under Sakhalin, and becomes flattened atop of the 660-km discontinuity beneath Korea, while only the tip of the slab reaches the lower TZ beneath the NEC. The TZ beneath the NCC does not show evidence of the slab stagnation.

Beschreibung: The fine-scale seismic structure of the central Mexico subduction zone is studied using moderate-sized (M4-6) intraslab earthquakes. Regional waveforms from the Mapping the Rivera Subduction Zone (MARS) seismic array are complicated and contain detailed information about the subduction zone structure, including evidence of lateral heterogeneity. This waveform information is used to model the structure of the subducted plates, particularly along the transition from flat to normal subduction, where recent studies have shown evidence for possible slab tearing along the eastern projection of the Orozco Fracture Zone (OFZ). The lateral extent of a thin ultra-slow velocity layer (USL) imaged atop the Cocos slab in recent studies along the Meso America Subduction Experiment array is examined here using MARS waveforms. We find an edge to this USL which is coincident with the western boundary of the projected OFZ region. Forward modeling of the 2D structure of the subducted Rivera and Cocos plates using a finite difference algorithm provides constraints on the velocity and geometry of each slab's seismic structure in this region and confirms the location of the USL edge. We propose that the Cocos slab is currently fragmenting into a North Cocos plate and a South Cocos plate along the projection of the OFZ, in agreement with observations of variable Cocos plate motion on either side of the OFZ. This tearing event may be a young analogy to the 10 Ma Rivera-Cocos plate boundary, and may be related to the slab rollback in central Mexico.

Beschreibung: Publication year: 2012 Source: Cell Reports Andre L. Samson, Anja S. Knaupp, Maithili Sashindranath, Rachael J. Borg, Amanda E.-L. Au, Elisa J. Cops, Helen M. Saunders, Stephen H. Cody, Catriona A. McLean, Cameron J. Nowell, Victoria A. Hughes, Stephen P. Bottomley, Robert L. Medcalf Cellular injury causes a myriad of processes that affect proteostasis. We describe nucleocytoplasmic coagulation (NCC), an intracellular disulfide-dependent protein crosslinking event occurring upon late-stage cell death that orchestrates the proteolytic removal of misfolded proteins. In vitro and in vivo models of neuronal injury show that NCC involves conversion of soluble intracellular proteins, including tubulin, into insoluble oligomers. These oligomers, also seen in human brain tissue following neurotrauma, act as a cofactor and substrate for the plasminogen-activating system. In plasminogen −/− mice, levels of misfolded β-tubulin were elevated and its clearance delayed following neurotrauma, demonstrating a requirement for plasminogen in the removal of NCC constituents. While additional in vivo studies will further dissect this phenomenon, our study clearly shows that NCC, a process analogous to the formation of thrombi, generates an aggregated protein scaffold that limits release of cellular components and recruits clearance mechanisms to the site of injury. Graphical abstract Graphical Abstract Highlights ► Cell death triggers the process of nucleocytoplasmic coagulation (NCC) ► NCC is the abrupt misfolding and disulfide crosslinking of intracellular proteins ► NCC-aggregated proteins facilitate plasmin formation and subsequent proteolysis ► Thus, NCC limits release and promotes proteolytic clearance of dead cell debris

Beschreibung: Publication year: 2012 Source: Cell Reports Jennifer S. Liu, Justin T. Farlow, Amanda K. Paulson, Mark A. Labarge, Zev J. Gartner Variability in signaling pathway activation between neighboring epithelial cells can arise from local differences in the microenvironment, noisy gene expression, or acquired genetic changes. To investigate the consequences of this cell-to-cell variability in signaling pathway activation on coordinated multicellular processes such as morphogenesis, we use DNA-programmed assembly to construct three-dimensional MCF10A microtissues that are mosaic for low-level expression of activated H-Ras. We find two emergent behaviors in mosaic microtissues: cells with activated H-Ras are basally extruded or lead motile multicellular protrusions that direct the collective motility of their wild-type neighbors. Remarkably, these behaviors are not observed in homogeneous microtissues in which all cells express the activated Ras protein, indicating that heterogeneity in Ras activity, rather than the total amount of Ras activity, is critical for these processes. Our results directly demonstrate that cell-to-cell variability in pathway activation within local populations of epithelial cells can drive emergent behaviors during epithelial morphogenesis. Graphical abstract Graphical Abstract Highlights ► Cell-to-cell variability in Ras activity triggers emergent behaviors ► Homogeneous and low-level Ras activation does not disrupt early morphogenesis ► Programmed assembly precisely controls the composition of mosaic epithelial tissues

Beschreibung: Publication year: 2012 Source: Cell Reports Chandan Shee, Janet L. Gibson, Susan M. Rosenberg Mutation hotspots and showers occur across phylogeny and profoundly influence genome evolution, yet the mechanisms that produce hotspots remain obscure. We report that DNA double-strand breaks (DSBs) provoke mutation hotspots via stress-induced mutation in Escherichia coli . With tet reporters placed 2 kb to 2 Mb (half the genome) away from an I- Sce I site, RpoS/DinB-dependent mutations occur maximally within the first 2 kb and decrease logarithmically to ∼60 kb. A weak mutation tail extends to 1 Mb. Hotspotting occurs independently of I-site/ tet- reporter-pair position in the genome, upstream and downstream in the replication path. RecD, which allows RecBCD DSB-exonuclease activity, is required for strong local but not long-distance hotspotting, indicating that double-strand resection and gap-filling synthesis underlie local hotspotting, and newly illuminating DSB resection in vivo. Hotspotting near DSBs opens the possibility that specific genomic regions could be targeted for mutagenesis, and could also promote concerted evolution (coincident mutations) within genes/gene clusters, an important issue in the evolution of protein functions. Graphical abstract Graphical Abstract Highlights ► Spontaneous mutation pathway in Escherichia coli causes hotpots at double-strand breaks ► Strong local (2–60 kb) hotspot mechanism double-strand resection and gap-fill ► Weak long-distance (1 Mb) mutagenesis by break-induced replication ► Break-induced replication and length of DNA-end resection in natural repair with sister chromosomes

Beschreibung: Publication year: 2012 Source: Cell Reports Aurélie Jory, Carlos Estella, Matt W. Giorgianni, Matthew Slattery, Todd R. Laverty, Gerald M. Rubin, Richard S. Mann Over 6,000 fragments from the genome of Drosophila melanogaster were analyzed for their ability to drive expression of GAL4 reporter genes in the third-instar larval imaginal discs. About 1,200 reporter genes drove expression in the eye, antenna, leg, wing, haltere, or genital imaginal discs. The patterns ranged from large regions to individual cells. About 75% of the active fragments drove expression in multiple discs; 20% were expressed in ventral, but not dorsal, discs (legs, genital, and antenna), whereas ∼23% were expressed in dorsal but not ventral discs (wing, haltere, and eye). Several patterns, for example, within the leg chordotonal organ, appeared a surprisingly large number of times. Unbiased searches for DNA sequence motifs suggest candidate transcription factors that may regulate enhancers with shared activities. Together, these expression patterns provide a valuable resource to the community and offer a broad overview of how transcriptional regulatory information is distributed in the Drosophila genome. Graphical abstract Graphical Abstract Highlights ► Over 6,000 fragments from the Drosophila genome were analyzed for enhancer ► About 1,200 fragments drove expression of GAL4 in at least one of six imaginal discs ► The lines greatly expand the number of GAL4 drivers available to the community

Beschreibung: Characterizing the vigor of magmatic activity in Yellowstone requires knowledge of the mechanisms and rates of heat transport between magma and the ground surface. We present results from a heat flow study in two vapor dominated, acid-sulfate thermal areas in the Yellowstone Caldera, the 0.11 km2 Obsidian Pool Thermal Area (OPTA) and the 0.25 km2 Solfatara Plateau Thermal Area (SPTA). Conductive heat flux through a low permeability layer capping large vapor reservoirs is calculated from soil temperature measurements at 〉600 locations and from laboratory measurements of soil properties. The conductive heat output is 3.6 ± 0.4 MW and 7.5 ± 0.4 MW from the OPTA and the SPTA, respectively. The advective heat output from soils is 1.3 ± 0.3 MW and 1.2 ± 0.3 MW from the OPTA and the SPTA, respectively and the heat output from thermal pools in the OPTA is 6.8 ± 1.4 MW. These estimates result in a total heat output of 11.8 ± 1.4 MW and 8.8 ± 0.4 MW from OPTA and SPTA, respectively. Focused zones of high heat flux in both thermal areas are roughly aligned with regional faults suggesting that faults in both areas serve as conduits for the rising acid vapor. Extrapolation of the average heat flux from the OPTA (103 ± 2 W·m−2) and SPTA (35 ± 3 W·m−2) to the ∼35 km2 of vapor dominated areas in Yellowstone yields 3.6 and 1.2 GW, respectively, which is less than the total heat output transported by steam from the Yellowstone Caldera as estimated by the chloride inventory method (4.0 to 8.0 GW).

Beschreibung: Using two recent high-resolution earthquake catalogs, I examine clustering in California seismicity by plotting the average rate of earthquakes as a function of both space and time from target events of M 2 to 5. Comparisons between pre- and post-target event activity can be used to resolve earthquake-to-earthquake triggering associated with target events of different magnitudes. The results are more complicated than predicted by computer simulations of earthquake triggering that begin with background events occurring at random times. In particular, at least some of the temporal clustering of seismicity at short scales (0.1 to 5 km) does not appear to be caused by local earthquake triggering, but instead reflects an underlying physical process that temporarily increases the seismicity rate, such as is often hypothesized to drive earthquake swarms. Earthquake triggering for M 〈 4.5 earthquakes is only resolvable in average seismicity rates at times less than about one day and to distances of less than about 10 km, and its linear density decreases as r−1.5 to r−2.5, significantly steeper than some previous studies have found.

Beschreibung: Although kinematic earthquake source inversions show dominantly pulse-like subshear rupture behavior, seismological observations, laboratory experiments and theoretical models indicate that earthquakes can operate with different rupture styles: either as pulses or cracks, that propagate at subshear or supershear speeds. The determination of rupture style and speed has important implications for ground motions and may inform about the state of stress and strength of active fault zones. We conduct 2D in-plane dynamic rupture simulations with a spectral element method to investigate the diversity of rupture styles on faults governed by velocity-and-state-dependent friction with dramatic velocity-weakening at high slip rate. Our rupture models are governed by uniform initial stresses, and are artificially initiated. We identify the conditions that lead to different rupture styles by investigating the transitions between decaying, steady state and growing pulses, cracks, sub-shear and super-shear ruptures as a function of background stress, nucleation size and characteristic velocity at the onset of severe weakening. Our models show that small changes of background stress or nucleation size may lead to dramatic changes of rupture style. We characterize the asymptotic properties of steady state and self-similar pulses as a function of background stress. We show that an earthquake may not be restricted to a single rupture style, but that complex rupture patterns may emerge that consist of multiple rupture fronts, possibly involving different styles and back-propagating fronts. We also demonstrate the possibility of a super-shear transition for pulse-like ruptures. Finally, we draw connections between our findings and recent seismological observations.

Beschreibung: We use boundary element methods to develop antiplane, strike-slip earthquake cycle models consisting of faulting in an elastic plate with possibly different thickness and stiffness on either side of the fault overlying a linear, Maxwell viscoelastic substrate. We show that isolated plate models that neglect the coupling of the plate to the underlying substrate might significantly overpredict the asymmetry in deformation across the fault. We also show that flow in a low-viscosity channel in the lower crust could significantly contribute to the asymmetry. Through a fully probabilistic scheme, we invert geodetic data across three strike-slip fault systems for effective elastic thickness and elastic stiffness on both sides of the fault using geological and geophysical constraints. For the Renun segment of the Great Sumatra fault, inversion results show the elastic layer on the east side is stiffer than the west side but the effective elastic thicknesses are not resolvable. For the Carrizo segment of the San Andreas fault, the inversion results slightly favor a thicker elastic layer on the east side (∼2.2 times) but stiffer layer on west side (∼1.2 times); however, uniform effective elastic thickness and stiffness cannot be ruled out. For the Aksay segment of the Altyn Tagh fault in northern Tibet, inversion results show the effective elastic crust of the Tarim Basin must be stiffer and thicker than the effective elastic crust of the Tibetan Plateau to the south, but the viscosity of a hypothesized mid-crustal Tibetan channel is not resolvable.

Beschreibung: Morphochronologic slip-rates on the Altyn Tagh Fault (ATF) along the southern front of the Pingding Shan at ∼90.5°E are determined by cosmogenic radionuclide (CRN) dating of seven offset terraces at two sites. The terraces are defined based upon morphology, elevation and dating, together with fieldwork and high-resolution satellite analysis. The majority of the CRN model ages fall within narrow ranges (

Beschreibung: Publication year: 2012 Source: Cell Reports Rachel Spokoini, Ofer Moldavski, Yaakov Nahmias, Jeremy L. England, Maya Schuldiner, Daniel Kaganovich The division of the S. cerevisiae budding yeast, which produces one mother cell and one daughter cell, is asymmetric with respect to aging. Remarkably, the asymmetry of yeast aging coincides with asymmetric inheritance of damaged and aggregated proteins by the mother cell. Here, we show that misfolded proteins are retained in the mother cell by being sequestered in juxtanuclear quality control compartment (JUNQ) and insoluble protein deposit (IPOD) inclusions, which are attached to organelles. Upon exposure to stress, misfolded proteins accumulate in stress foci that must be disaggregated by Hsp104 in order to be degraded or processed to JUNQ and IPOD. Cells that fail to deliver aggregates to an inclusion pass on aggregates to subsequent generations. Graphical abstract Graphical Abstract Highlights ► Misfolded proteins form stress foci, JUNQ inclusions, or IPOD inclusions ► JUNQs and IPODs are attached to organelles ► JUNQ and IPOD inclusions are asymmetrically inherited during cell division ► Hsp104 is required for disaggregation of stress foci

Beschreibung: Publication year: 2012 Source: Cell Reports Ellen M. van Beek, Julian Alvarez Zarate, Robin van Bruggen, Karin Schornagel, Anton T.J. Tool, Takashi Matozaki, Georg Kraal, Dirk Roos, Timo K. van den Berg The phagocyte NADPH oxidase mediates oxidative microbial killing in granulocytes and macrophages. However, because the reactive oxygen species produced by the NADPH oxidase can also be toxic to the host, it is essential to control its activity. Little is known about the endogenous mechanism(s) that limits NADPH oxidase activity. Here, we demonstrate that the myeloid-inhibitory receptor SIRPα acts as a negative regulator of the phagocyte NADPH oxidase. Phagocytes isolated from SIRPα mutant mice were shown to have an enhanced respiratory burst. Furthermore, overexpression of SIRPα in human myeloid cells prevented respiratory burst activation. The inhibitory effect required interactions between SIRPα and its natural ligand, CD47, as well as signaling through the SIRPα cytoplasmic immunoreceptor tyrosine-based inhibitory motifs. Suppression of the respiratory burst by SIRPα was caused by a selective repression of gp91 phox expression, the catalytic component of the phagocyte NADPH oxidase complex. Thus, SIRPα can limit gp91 phox expression during myeloid development, thereby controlling the magnitude of the respiratory burst in phagocytes. Graphical abstract Graphical Abstract Highlights ► Activity of the phagocyte NADPH oxidase is restricted by SIRPα ► This involves SIRP-CD47 interactions and signaling through the SIRPα immunoreceptor tyrosine-based inhibitory motifs ► SIRPα signaling represses expression of the catalytic NADPH oxidase subunit gp91 phox

Beschreibung: Publication year: 2012 Source: Cell Reports Ping Yin, Dong Deng, Chuangye Yan, Xiaojing Pan, Jianzhong Jeff Xi, Nieng Yan, Yigong Shi The transcription activator-like (TAL) effector targets specific host promoter through its central DNA-binding domain, which comprises multiple tandem repeats (TALE repeats). Recent structural analyses revealed that the TALE repeats form a superhelical structure that tracks along the forward strand of the DNA duplex. Here, we demonstrate that TALE repeats specifically recognize a DNA-RNA hybrid where the DNA strand determines the binding specificity. The crystal structure of a designed TALE in complex with the DNA-RNA hybrid was determined at a resolution of 2.5 Å. Although TALE repeats are in direct contact with only the DNA strand, the phosphodiester backbone of the RNA strand is inaccessible by macromolecules such as RNases. Consistent with this observation, sequence-specific recognition of an HIV-derived DNA-RNA hybrid by an engineered TALE efficiently blocked RNase H-mediated degradation of the RNA strand. Our study broadens the utility of TALE repeats and suggests potential applications in processes involving DNA replication and retroviral infections. Graphical abstract Graphical Abstract Highlights ► TALE repeats specifically bind to DNA-RNA hybrids ► The sequence of DNA determines binding specificity by TALE repeats ► TALE repeats protect DNA-RNA hybrids from RNase H degradation ► TALEs may be used to fight retroviral infection and modulate transcription

Beschreibung: Publication year: 2012 Source: Cell Reports Michael Hiller, Bruce T. Schaar, Vahan B. Indjeian, David M. Kingsley, Lee R. Hagey, Gill Bejerano Genotype-phenotype mapping is hampered by countless genomic changes between species. We introduce a computational “forward genomics” strategy that—given only an independently lost phenotype and whole genomes—matches genomic and phenotypic loss patterns to associate specific genomic regions with this phenotype. We conducted genome-wide screens for two metabolic phenotypes. First, our approach correctly matches the inactivated Gulo gene exactly with the species that lost the ability to synthesize vitamin C. Second, we attribute naturally low biliary phospholipid levels in guinea pigs and horses to the inactivated phospholipid transporter Abcb4. Human ABCB4 mutations also result in low phospholipid levels but lead to severe liver disease, suggesting compensatory mechanisms in guinea pig and horse. Our simulation studies, counts of independent changes in existing phenotype surveys, and the forthcoming availability of many new genomes all suggest that forward genomics can be applied to many phenotypes, including those relevant for human evolution and disease. Graphical abstract Graphical Abstract Highlights ► Matching independent phenotypic losses with ancestral genomic information erosion ► Gulo gene loss uniquely matches to “loss of vitamin C synthesis” in mammals ► Abcb4 gene loss matches “low biliary phospholipid levels” in guinea pig & horse ► Broad applicability of the approach from simulation and phenotype measurements

Beschreibung: The spectral induced polarization (SIP) signature of soil contaminated with organic pollutant was studied. Using a flow column experiment, the effect of crystal violet (CV, a polar organic pollutant) on the temporal change of the SIP response over a broad frequency range (1 mHz to 45 KHz) was determined. Complimentary measurements were used to determine the effect of CV on the chemical composition of both the pore water and the solid surface. In addition, analysis of the experimental results was carried out by using both chemical complexation and induced polarization models. Our results shows that adsorption of CV to the mineral surface resulted in release of inorganic ions to the soil solution, increasing the solution electrical conductivity and therefore also the real part of the complex conductivity. Despite the increase in the real part of the complex conductivity, the imaginary part of the complex conductivity decreased with increasing concentration of adsorbed CV. Using the Revil induced polarization model, we were able to show that the contribution of the adsorbed CV to the polarization of the soil is negligible, and that the main process affecting the polarization is the decrease in the density of the inorganic surface species. The results of this study can be used to better interpret SIP signature of soils contaminated by organic compounds.

Beschreibung: Hydrothermal convection in the Taupo Volcanic Zone (TVZ), New Zealand, is driven by heat extracted at the brittle-ductile transition, which is in turn supplied by magmatic intrusion of the lower-crust. We present a numerical model that approximates this circulation in a statistical sense, being constrained by TVZ dimensions and mean thermal properties, but incorporating a permeability distribution of arbitrary heterogeneity. A particle tracking methodology that accounts for advective and dispersive transport is introduced, and ensembles of several tens of thousands of flow paths are constructed for each of the modeled geothermal systems. These flow paths reveal the nature of mass recharge in multicell convective systems, and suggest the ages of waters in TVZ geothermal systems vary between 5 and 50 kyr. Flow path ensembles are used to delineate catchments for each of the modeled systems and a method for calculating their areas is introduced. Catchment area is shown to be proportional to system heat output and area, which is consistent with a 1-D analytical model of heat and mass transfer. As an approximation to catchments delineated by particle tracking, we outline a method of Voronoi tessellation based on the positions and heat outputs of geothermal systems. This method is used to delineate catchments for geothermal systems in the TVZ and Iceland.

Beschreibung: Publication year: 2012 Source: Cell Reports Jian Zhu, Gaurav D. Gaiha, Sinu P. John, Thomas Pertel, Christopher R. Chin, Geng Gao, Hongjing Qu, Bruce D. Walker, Stephen J. Elledge, Abraham L. Brass HIV-1 depends on many host factors for propagation. Other host factors, however, antagonize HIV-1 and may have profound effects on viral activation. Curing HIV-1 requires the reduction of latent viral reservoirs that remain in the face of antiretroviral therapy. Using orthologous genetic screens, we identified bromodomain containing 4 (BRD4) as a negative regulator of HIV-1 replication. Antagonism of BRD4, via RNA interference or with a small molecule inhibitor, JQ1, both increased proviral transcriptional elongation and alleviated HIV-1 latency in cell-line models. In multiple instances, JQ1, when used in combination with the NF-κB activators Prostratin or PHA, enhanced the in vitro reactivation of latent HIV-1 in primary T cells. These data are consistent with a model wherein BRD4 competes with the virus for HIV-1 dependency factors (HDFs) and suggests that combinatorial therapies that activate HDFs and antagonize HIV-1 competitive factors may be useful for curing HIV-1 infection. Graphical abstract Graphical Abstract Highlights ► BRD4 depletion or inhibition with JQ1 increases HIV-1 replication and gene expression ► BRD4 inhibition increases Tat-dependent transcriptional elongation and Tat–PTEF-b association ► BRD4 inhibition alleviates HIV-1 latency in cell-line models ► JQ1 with HDF activators enhances HIV-1 replication in primary and latently infected T cells

Beschreibung: Publication year: 2012 Source: Cell Reports Stefanie Reissmann, Lukasz A. Joachimiak, Bryan Chen, Anne S. Meyer, Anthony Nguyen, Judith Frydman The eukaryotic chaperonin TRiC/CCT uses ATP cycling to fold many essential proteins that other chaperones cannot fold. This 1 MDa hetero-oligomer consists of two identical stacked rings assembled from eight paralogous subunits, each containing a conserved ATP-binding domain. Here, we report a dramatic asymmetry in the ATP utilization cycle of this ring-shaped chaperonin, despite its apparently symmetric architecture. Only four of the eight different subunits bind ATP at physiological concentrations. ATP binding and hydrolysis by the low-affinity subunits is fully dispensable for TRiC function in vivo . The conserved nucleotide-binding hierarchy among TRiC subunits is evolutionarily modulated through differential nucleoside contacts. Strikingly, high- and low-affinity subunits are spatially segregated within two contiguous hemispheres in the ring, generating an asymmetric power stroke that drives the folding cycle. This unusual mode of ATP utilization likely serves to orchestrate a directional mechanism underlying TRiC/CCT's unique ability to fold complex eukaryotic proteins. Graphical abstract Graphical Abstract Highlights ► The eight paralogous TRiC subunits display hierarchical ATP occupancy ► Conservation of nucleoside contacts among TRiC orthologs mirrors ATP affinity ► ATP binding and hydrolysis in the low-affinity subunits are dispensable for life ► ATP usage segregates asymmetrically into two hemispheres of the chaperonin ring

Beschreibung: Publication year: 2012 Source: Cell Reports Gil Diamant, Liat Amir-Zilberstein, Yuki Yamaguchi, Hiroshi Handa, Rivka Dikstein NF-κB is central for immune response and cell survival, and its deregulation is linked to chronic inflammation and cancer through poorly defined mechanisms. IκBα and A20 are NF-κB target genes and negative feedback regulators. Upon their activation by NF-κB, DSIF is recruited, P-TEFb is released, and their elongating polymerase II (Pol II) C-terminal domain (CTD) remains hypophosphorylated. We show that upon DSIF knockdown, mRNA levels of a subset of NF-κB targets are not diminished; yet much less IκBα and A20 protein are synthesized, and NF-κB activation is abnormally prolonged. Further analysis of IκBα and A20 mRNA revealed that a significant portion is uncapped, unspliced, and retained in the nucleus. Interestingly, the Spt5 C-terminal repeat (CTR) domain involved in elongation stimulation through P-TEFb is dispensable for IκBα and A20 regulation. These findings assign a function for DSIF in cotranscriptional mRNA processing when elongating Pol II is hypophosphorylated and define DSIF as part of the negative feedback regulation of NF-κB. Graphical abstract Graphical Abstract Highlights ► IκBα and A20, negative feedback regulators of NF-κB, have unusual elongation control ► Upon activation of IκBα and A20, DSIF is recruited, and the Pol II CTD remains hypophosphorylated ► DSIF is selectively required for IκBα and A20 mRNA processing and proper NF-κ B signaling ► DSIF’s CTR domain involved in elongation stimulation is dispensable for this function

Beschreibung: Publication year: 2012 Source: Cell Reports Samantha A. Morris, Yu Guo, Magdalena Zernicka-Goetz Plasticity is a well-known feature of mammalian development, and yet very little is known about its underlying mechanism. Here, we establish a model system to examine the extent and limitations of developmental plasticity in living mouse embryos. We show that halved embryos follow the same strict clock of developmental transitions as intact embryos, but their potential is not equal. We have determine that unless a minimum of four pluripotent cells are established before implantation, development will arrest. This failure can be rescued by modulating Fgf and Wnt signaling to enhance the pluripotent cell number, allowing the generation of monozygotic twins, which is an otherwise rare phenomenon. Knowledge of the minimum pluripotent-cell number required for development to birth, as well as the different potentials of blastomeres, allowed us to establish a protocol for splitting an embryo into one part that develops to adulthood and another that provides embryonic stem cells for that individual. Graphical abstract Graphical Abstract Highlights ► Half embryos follow the same clock as intact embryos, but their potential is not equal ► To support development, four pluripotent cells must be generated before implantation ► Fgf/Wnt signal modulation enhances pluripotency to rescue half-embryo development ► ESCs and a viable mouse can be derived from a single embryo with high efficiency

Beschreibung: Publication year: 2012 Source: Cell Reports Pol Margalef, Vanessa Fernández-Majada, Alberto Villanueva, Ricard Garcia-Carbonell, Mar Iglesias, Laura López, María Martínez-Iniesta, Jordi Villà-Freixa, Mari Carmen Mulero, Montserrat Andreu, Ferran Torres, Marty W. Mayo, Anna Bigas, Lluis Espinosa Nuclear IKKα regulates gene transcription by phosphorylating specific substrates and has been linked to cancer progression and metastasis. However, the mechanistic connection between tumorigenesis and IKKα activity remains poorly understood. We have now analyzed 288 human colorectal cancer samples and found a significant association between the presence of nuclear IKK and malignancy. Importantly, the nucleus of tumor cells contains an active IKKα isoform with a predicted molecular weight of 45 kDa (p45-IKKα) that includes the kinase domain but lacks several regulatory regions. Active nuclear p45-IKKα forms a complex with nonactive IKKα and NEMO that mediates phosphorylation of SMRT and histone H3. Proteolytic cleavage of FL-IKKα into p45-IKKα is required for preventing the apoptosis of CRC cells in vitro and sustaining tumor growth in vivo. Our findings identify a potentially druggable target for treating patients with advance refractory CRC. Graphical abstract Graphical Abstract Highlights ► A truncated active form of IKKα is found in colorectal cancer cells ► Nuclear complex containing p45-IKKα phosphorylates SMRT and histone H3 ► Cleavage of IKK(alpha) into p45-IKKα is required for cancer cell growth

Beschreibung: We develop a global-scale P wave velocity model (LLNL-G3Dv3) designed to accurately predict seismic travel times at regional and teleseismic distances simultaneously. The model provides a new image of Earth's interior, but the underlying practical purpose of the model is to provide enhanced seismic event location capabilities. The LLNL-G3Dv3 model is based on ∼2.8 million P and Pn arrivals that are re-processed using our global multiple-event locator called Bayesloc. We construct LLNL-G3Dv3 within a spherical tessellation based framework, allowing for explicit representation of undulating and discontinuous layers including the crust and transition zone layers. Using a multiscale inversion technique, regional trends as well as fine details are captured where the data allow. LLNL-G3Dv3 exhibits large-scale structures including cratons and superplumes as well numerous complex details in the upper mantle including within the transition zone. Particularly, the model reveals new details of a vast network of subducted slabs trapped within the transition beneath much of Eurasia, including beneath the Tibetan Plateau. We demonstrate the impact of Bayesloc multiple-event location on the resulting tomographic images through comparison with images produced without the benefit of multiple-event constraints (single-event locations). We find that the multiple-event locations allow for better reconciliation of the large set of direct P phases recorded at 0–97° distance and yield a smoother and more continuous image relative to the single-event locations. Travel times predicted from a 3-D model are also found to be strongly influenced by the initial locations of the input data, even when an iterative inversion/relocation technique is employed.

Beschreibung: Providing high-fidelity paleointensity estimates is pivotal to identify temporal fluctuation of ancient geomagnetic field. Despite abundance of available paleointensity data in Japan, only dozen of them satisfy modern standards with systematic alteration checks. High-fidelity paleointensity estimates were obtained from historic andesitic lavas collected from Mt. Aso (Ojodake, ≈700 BC), Mt. Kirishima (Iwoyama, AD 1768; Ohachi, AD 1235), and Mt. Sakurajima (An-ei, AD 1779; Nabeyama, AD 764–766). Variation of geomagnetic field intensity is distinctively different from the prediction of global models (ARCH3k.1, CALS3k.3, CALS3k.4, CALS7k.2, and CALS10k.1b) for the past 4000 years in Japan. The compilation of high-fidelity Thellier data set in Japan showed two obvious high intensities at AD 590–765 and AD 1330–1435 and one low intensity at ≈700 BC. These time intervals with anomalous high/low intensities are nearly identical to three of the four potential archeomagnetic jerks recognized from the European archeomagnetic data, implying that the archeomagnetic jerks were global (or at least northern hemispheric) features. To improve the poor representation of regional geomagnetic field variation with respect to the global model prediction, more high-fidelity paleointensity determinations are required in East Asia.

Beschreibung: Publication year: 2012 Source: Cell Reports Adrian W.R. Serohijos, Zilvinas Rimas, Eugene I. Shakhnovich The consistent observation across all kingdoms of life that highly abundant proteins evolve slowly demonstrates that cellular abundance is a key determinant of protein evolutionary rate. However, other empirical findings, such as the broad distribution of evolutionary rates, suggest that additional variables determine the rate of protein evolution. Here, we report that under the global selection against the cytotoxic effects of misfolded proteins, folding stability (Δ G ), simultaneous with abundance, is a causal variable of evolutionary rate. Using both theoretical analysis and multiscale simulations, we demonstrate that the anticorrelation between the premutation Δ G and the arising mutational effect (ΔΔ G ), purely biophysical in origin, is a necessary requirement for abundance–evolutionary rate covariation. Additionally, we predict and demonstrate in bacteria that the strength of abundance–evolutionary rate correlation depends on the divergence time separating reference genomes. Altogether, these results highlight the intrinsic role of protein biophysics in the emerging universal patterns of molecular evolution. Graphical abstract Graphical Abstract Highlights ► Protein stability and abundance mutually determine its evolutionary rate ► Coupling of stability changes with wild-type stability affects rate of evolution ► Abundance–evolutionary rate covariation depends on divergence time

Beschreibung: We present an improved rendition of the geodetic velocity and strain fields in Sicily and southern Calabria obtained through the analysis of 18 years of GPS observations from continuous and survey station networks. The dense spatial coverage of geodetic data provides precise quantitative estimates of previously established first-order active kinematic features, including: i) a narrow east-west-elongated belt of contraction (∼1–1.5 mm/yr) extending offshore northern Sicily from Ustica to Stromboli across the Aeolian Islands; ii) a narrow east-west-trending contractional belt located along the northern rim of the Hyblean Plateau in southern Sicily, with shortening at up to 4.4 mm/yr; iii) right motion (∼3.6 mm/yr) on the Aeolian-Tindari-Letojanni fault (ATLF) system, a main shear zone extending from the Aeolian Islands to the Ionian coast of Sicily, with significant transpression and transtension partitioned between discrete sectors of the fault; iv) transtension (∼1 mm/yr) across the Sicily Channel between Sicily and North Africa. We use geodetic observations coupled to geological constraints to better elucidate the interplay of crustal blocks revealed in the investigated area. In particular, we focus on the ATLF, which forms the primary boundary between the Sicilian and Calabrian blocks. The ATLF juxtaposes north-south contraction between Sicily and the Tyrrhenian block with northwest-southeast extension in northeastern Sicily and Calabria. Contraction between Sicily and Tyrrhenian blocks probably arises from the main Europe-Nubia convergence, although Sicily has a component of lateral motion away from Nubia. We found that convergence is not restricted to the northern offshore, as commonly believed, but is widely accommodated between the frontal belt and the northern rim of the Hyblean foreland in southern Sicily. Geodetic data also indicate that active right shear on the ATLF occurs to the southeast of the mapped fault array in northern Sicily, suggesting the fault cuts through till the Ionian coast of the island. The small geodetic divergence between the Hyblean and Apulian blocks rimming on both sides the Calabria block and subjacent Ionian slab, coupled with marine geophysical evidences in the Ionian Sea lends credit to the proposed deep root of the ATLF and to a fragmentation of the Ionian domain.

Beschreibung: New high-precision U/Pb zircon geochronology from the Oman-United Arab Emirates (U.A.E.) ophiolite provides insight into the timing and duration of magmatism and the tectonic setting during formation of the lower crust. The new data come from a well-preserved and exposed crustal section in the center of the Wadi Tayin massif. Single grain and grain fragment 206Pb/238U dates from upper-level gabbros, tonalites/trondhjemites and gabbroic pegmatites, corrected for initial Th exclusion, range from 112.55 ± 0.21 to 95.50 ± 0.17 Ma, with most data clustered between 96.40 ± 0.17 to 95.50 ± 0.17 Ma. Zircon dates from upper-level gabbros are most consistent with the ophiolite forming at a fast spreading ridge with half-rates of 50–100 km/Ma. Dates from tonalites/trondhjemites and from a gabbroic pegmatite associated with a wehrlite intrusion overlap with dates from adjacent upper-level gabbros, suggesting that any age differences between these three magmatic series are smaller than the analytical uncertainties or intrasample variability in the dates. Three of the dated upper-level gabbros and a single gabbroic pegmatite from the base of the crust have 〉1 Ma intrasample variability in single grain dates, suggesting assimilation of older crust during the formation or crystallization of the magmas. Whole rock εNd(t) of seven samples, including the upper-level gabbros with variable zircon dates, have tightly clustered initial values ranging from εNd(96 Ma) = 7.59 ± 0.23 to 8.28 ± 0.31. The εNd values are similar to those from other gabbros within the ophiolite, suggesting that any assimilated material had a similar isotopic composition to primitive basaltic magmas. The new dates suggest that the studied section formed at a fast spreading mid-ocean ridge between ∼96.4–95.5 Ma. The large intrasample variability in zircon dates in some samples is unexpected in this setting, and may be related to propagation of a younger ridge into older oceanic lithosphere.