Multiple endocytic pathways of G protein-coupled receptors delineated by GIT1 sensitivity.

Recently, we identified a GTPase-activating protein for the ADP ribosylation factor family of small GTP-binding proteins that we call GIT1. This protein initially was identified as an interacting partner for the G protein-coupled receptor kinases, and its overexpression was found to affect signaling and internalization of the prototypical beta(2)-adrenergic receptor. Here, we report that GIT1 overexpression regulates internalization of numerous, but not all, G protein-coupled receptors. The specificity of the GIT1 effect is not related to the type of G protein to which a receptor couples, but rather to the endocytic route it uses. GIT1 only affects the function of G protein-coupled receptors that are internalized through the clathrin-coated pit pathway in a beta-arrestin- and dynamin-sensitive manner. Furthermore, the GIT1 effect is not limited to G protein-coupled receptors because overexpression of this protein also affects internalization of the epidermal growth factor receptor. However, constitutive agonist-independent internalization is not regulated by GIT1, because transferrin uptake is not affected by GIT1 overexpression. Thus, GIT1 is a protein involved in regulating the function of signaling receptors internalized through the clathrin pathway and can be used as a diagnostic tool for defining the endocytic pathway of a receptor

The Dzhungarian fault: Late Quaternary tectonics and slip rate of a major right-lateral strike-slip fault in the northern Tien Shan region

The Dzhungarian strike-slip fault of Central Asia is one of a series of long, NW-SE right-lateral strike-slip faults that are characteristic of the northern Tien Shan region and extends over 300 km from the high mountains into the Kazakh Platform. Our field-based and satellite observations reveal that the Dzhungarian fault can be characterized by three 100 km long sections based on variation in strike direction. Through morphological analysis of offset streams and alluvial fans, and through optically stimulated luminescence dating, we find that the Dzhungarian fault has a minimum average late Quaternary slip rate of 2.2 ± 0.8 mm/yr and accommodates N-S shortening related to the India-Eurasia collision. This shortening may also be partly accommodated by counterclockwise rotation about a vertical axis. Evidence for a possible paleo-earthquake rupture indicates that earthquakes up to at least Mw 7 can be associated with just the partitioned component of reverse slip on segments of the central section of the fault up to 30 km long. An event rupturing longer sections of the Dzhungarian fault has the potential to generate greater magnitude earthquakes (Mw 8); however, long time periods (e.g., thousands of years) are expected in order to accumulate enough strain to generate such earthquakes.We thank the Royal Society International Travel Grant, Mike Coward Fund of the Geological Society of London, Percy Sladen Fund of the Linnean Society, The Gilchrist Educational Trust, and the Earth and Space Foundation for their support in funding this project. GEC’s doctoral studentship is funded by the National Environmental Research Council through NCEO, COMET, and the NERC-ESRC funded Earthquakes without Frontiers (EWF) Project. RTW is supported by a University Research Fellowship awarded by the Royal Society.This is the final version of the article, originally published in the Journal of Geophysical Research: Solid Earth. It is also available from Wiley at http://onlinelibrary.wiley.com/doi/10.1002/jgrb.50367/abstract. © 2013. American Geophysical Unio

Seismotectonics and rupture process of the MW 7.1 2011 Van reverse-faulting earthquake, eastern Turkey, and implications for hazard in regions of distributed shortening

The 2011 October 23 MW 7.1 Van earthquake in eastern Turkey caused ~600 deaths and caused widespread damage and economic loss. The seismogenic rupture was restricted to 10-25 km in depth, but aseismic surface creep, coincident with outcrop fault exposures, was observed in the hours to months after the earthquake. We combine observations from radar interferometry, seismology, geomorphology and Quaternary dating to investigate the geological slip rate and seismotectonic context of the Van earthquake, and assess the implications for continuing seismic hazard in the region. Transient post-seismic slip on the upper Van fault started immediately following the earthquake, and decayed over a period of weeks; it may not fully account for our long-term surface slip-rate estimate of ≥ 0.5 mm yr-1. Post-seismic slip on the Bostaniçi splay fault initiated several days to weeks after the main shock, and we infer that it may have followed the MW 5.9 aftershock on the 9th November. The Van earthquake shows that updip segmentation can be important in arresting seismic ruptures on dip-slip faults. Two large, shallow aftershocks show that the upper 10 km of crust can sustain significant earthquakes, and significant slip is observed to have reached the surface in the late Quaternary, so there may be a continuing seismic hazard from the upper Van fault and the associated splay. The wavelength of folding in the hanging wall of the Van fault is dominated by the structure in the upper 10 km of the crust, masking the effect of deeper seismogenic structures. Thus, models of subsurface faulting based solely on surface folding and faulting in regions of reverse faulting may underestimate the full depth extent of seismogenic structures in the region. In measuring the cumulative post-seismic offsets to anthropogenic structures, we show that Structure-from-Motion can be rapidly deployed to create snapshots of postseismic displacement.We also demonstrate the utility of declassified Corona mission imagery (1960s-1970s) for geomorphic mapping in areas where recent urbanization has concealed the geomorphic markers

A viral CTL escape mutation leading to immunoglobulin-like transcript 4-mediated functional inhibition of myelomonocytic cells

Viral mutational escape can reduce or abrogate recognition by the T cell receptor (TCR) of virus-specific CD8+ T cells. However, very little is known about the impact of cytotoxic T lymphocyte (CTL) epitope mutations on interactions between peptide–major histocompatibility complex (MHC) class I complexes and MHC class I receptors expressed on other cell types. Here, we analyzed a variant of the immunodominant human leukocyte antigen (HLA)-B2705–restricted HIV-1 Gag KK10 epitope (KRWIILGLNK) with an L to M amino acid substitution at position 6 (L6M), which arises as a CTL escape variant after primary infection but is sufficiently immunogenic to elicit a secondary, de novo HIV-1–specific CD8+ T cell response with an alternative TCR repertoire in chronic infection. In addition to altering recognition by HIV-1–specific CD8+ T cells, the HLA-B2705–KK10 L6M complex also exhibits substantially increased binding to the immunoglobulin-like transcript (ILT) receptor 4, an inhibitory MHC class I–specific receptor expressed on myelomonocytic cells. Binding of the B2705–KK10 L6M complex to ILT4 leads to a tolerogenic phenotype of myelomonocytic cells with lower surface expression of dendritic cell (DC) maturation markers and co-stimulatory molecules. These data suggest a link between CTL-driven mutational escape, altered recognition by innate MHC class I receptors on myelomonocytic cells, and functional impairment of DCs, and thus provide important new insight into biological consequences of viral sequence diversificatio

The egiin davaa prehistoric rupture, central mongolia: A large magnitude normal faulting earthquake on a reactivated fault with little cumulative slip located in a slowly deforming intraplate setting

The prehistoric Egiin Davaa earthquake rupture is well-preserved in late Quaternary deposits within the Hangay Mountains of central Mongolia. The rupture is expressed by a semicontinuous 80 km-long topographic scarp. Geomorphological reconstructions reveal a relatively constant scarp height of 4-4.5 m and a NW-directed slip vector. Previous researchers have suggested that the scarp's exceptional geomorphological preservation indicates that it may correspond to an earthquake that occurred in the region c. 500 years ago. However, we constrain the last rupture to have been at least 4 ka ago from morphological dating and < 7.4 ka ago based on radiocarbon dating from one of two palaeoseismic trenches. Our study shows that discrete earthquake ruptures, along with details such as the locations of partially infilled fissures, can be preserved for periods well in excess of 1000 years in the interior of Asia, providing an archive of fault movements that can be directly read from the Earth's surface over a timescale appropriate for the study of slowly deforming continental interiors. The Egiin Davaa rupture involved c. 8 m of slip which, along with the observations that it is largely unsegmented along its length and that the ratio of cumulative slip (c. 250 m) to fault length (c. 80 km) is small, suggests relatively recent reactivation of a pre-existing geological structure

Hsf1 and Hsp90 orchestrate temperature-dependent global transcriptional remodelling and chromatin architecture in Candida albicans

This is the final version. Available from Nature Research via the DOI in this record.RNA-sequencing data sets are available at ArrayExpress (www.ebi.ac.uk) under accession code E-MTAB-4075. ChIP-seq data sets are available at the NCBI SRA database (http://www.ncbi.nlm.nih.gov) under accession code SRP071687. The authors declare that all other data supporting the findings of this study are available within the article and its supplementary information files, or from the corresponding author upon request.Fever is a universal response to infection, and opportunistic pathogens such as Candida albicans have evolved complex circuitry to sense and respond to heat. Here we harness RNA-seq and ChIP-seq to discover that the heat shock transcription factor, Hsf1, binds distinct motifs in nucleosome-depleted promoter regions to regulate heat shock genes and genes involved in virulence in C. albicans. Consequently, heat shock increases C. albicans host cell adhesion, damage and virulence. Hsf1 activation depends upon the molecular chaperone Hsp90 under basal and heat shock conditions, but the effects are opposite and in part controlled at the level of Hsf1 expression and DNA binding. Finally, we demonstrate that Hsp90 regulates global transcription programs by modulating nucleosome levels at promoters of stress-responsive genes. Thus, we describe a mechanism by which C. albicans responds to temperature via Hsf1 and Hsp90 to orchestrate gene expression and chromatin architecture, thereby enabling thermal adaptation and virulence.Wellcome TrustCanadian Institutes of Health ResearchCanadian Institutes of Health ResearchBiotechnology and Biological Sciences Research Council (BBSRC)European Research Council (ERC)Science and Technology Development Fund of Macau S.A.R (FDCT)Research and Development Administrative Office of the University of MacauNational Institutes of Health (NIH

Safety, tumor trafficking and immunogenicity of chimeric antigen receptor (CAR)-T cells specific for TAG-72 in colorectal cancer.

BackgroundT cells engineered to express chimeric antigen receptors (CARs) have established efficacy in the treatment of B-cell malignancies, but their relevance in solid tumors remains undefined. Here we report results of the first human trials of CAR-T cells in the treatment of solid tumors performed in the 1990s.MethodsPatients with metastatic colorectal cancer (CRC) were treated in two phase 1 trials with first-generation retroviral transduced CAR-T cells targeting tumor-associated glycoprotein (TAG)-72 and including a CD3-zeta intracellular signaling domain (CART72 cells). In trial C-9701 and C-9702, CART72 cells were administered in escalating doses up to 1010 total cells; in trial C-9701 CART72 cells were administered by intravenous infusion. In trial C-9702, CART72 cells were administered via direct hepatic artery infusion in patients with colorectal liver metastases. In both trials, a brief course of interferon-alpha (IFN-α) was given with each CART72 infusion to upregulate expression of TAG-72.ResultsFourteen patients were enrolled in C-9701 and nine in C-9702. CART72 manufacturing success rate was 100% with an average transduction efficiency of 38%. Ten patients were treated in CC-9701 and 6 in CC-9702. Symptoms consistent with low-grade, cytokine release syndrome were observed in both trials without clear evidence of on target/off tumor toxicity. Detectable, but mostly short-term (≤14&nbsp;weeks), persistence of CART72 cells was observed in blood; one patient had CART72 cells detectable at 48&nbsp;weeks. Trafficking to tumor tissues was confirmed in a tumor biopsy from one of three patients. A subset of patients had 111Indium-labeled CART72 cells injected, and trafficking could be detected to liver, but T cells appeared largely excluded from large metastatic deposits. Tumor biomarkers carcinoembryonic antigen (CEA) and TAG-72 were measured in serum; there was a precipitous decline of TAG-72, but not CEA, in some patients due to induction of an interfering antibody to the TAG-72 binding domain of humanized CC49, reflecting an anti-CAR immune response. No radiologic tumor responses were observed.ConclusionThese findings demonstrate the relative safety of CART72 cells. The limited persistence supports the incorporation of co-stimulatory domains in the CAR design and the use of fully human CAR constructs to mitigate immunogenicity

Atmospheric Evolution

Earth's atmosphere has evolved as volatile species cycle between the atmosphere, ocean, biomass and the solid Earth. The geochemical, biological and astrophysical processes that control atmospheric evolution are reviewed from an "Earth Systems" perspective, with a view not only to understanding the history of Earth, but also to generalizing to other solar system planets and exoplanets.Comment: 34 pages, 3 figures, 2 tables. Accepted as a chapter in "Encyclopaedia of Geochemistry", Editor Bill White, Springer-Nature, 201

Deletion of the gabra2 gene results in hypersensitivity to the acute effects of ethanol but does not alter ethanol self administration

Human genetic studies have suggested that polymorphisms of the GABRA2 gene encoding the GABA(A) α2-subunit are associated with ethanol dependence. Variations in this gene also convey sensitivity to the subjective effects of ethanol, indicating a role in mediating ethanol-related behaviours. We therefore investigated the consequences of deleting the α2-subunit on the ataxic and rewarding properties of ethanol in mice. Ataxic and sedative effects of ethanol were explored in GABA(A) α2-subunit wildtype (WT) and knockout (KO) mice using a Rotarod apparatus, wire hang and the duration of loss of righting reflex. Following training, KO mice showed shorter latencies to fall than WT littermates under ethanol (2 g/kg i.p.) in both Rotarod and wire hang tests. After administration of ethanol (3.5 g/kg i.p.), KO mice took longer to regain the righting reflex than WT mice. To ensure the acute effects are not due to the gabra2 deletion affecting pharmacokinetics, blood ethanol concentrations were measured at 20 minute intervals after acute administration (2 g/kg i.p.), and did not differ between genotypes. To investigate ethanol's rewarding properties, WT and KO mice were trained to lever press to receive increasing concentrations of ethanol on an FR4 schedule of reinforcement. Both WT and KO mice self-administered ethanol at similar rates, with no differences in the numbers of reinforcers earned. These data indicate a protective role for α2-subunits, against the acute sedative and ataxic effects of ethanol. However, no change was observed in ethanol self administration, suggesting the rewarding effects of ethanol remain unchange

Rapid mantle-driven uplift along the Angolan margin in the late Quaternary

Mantle flow can cause the Earth’s surface to uplift and subside, but the rates and durations of these motions are, in general, poorly resolved due to the difficulties in making measurements of relatively small vertical movements (hundreds of metres) over sufficiently large distances (about 1,000 km). Here we examine the effect of mantle upwelling through a study of Quaternary uplift along the coast of Angola. Using both optically stimulated luminescence on sediment grains, and radiocarbon dating of fossil shells, we date a 25 m coastal terrace at about 45 thousand years old, when sea level was about 75 m lower than today, indicating a rapid uplift rate of 1.8–2.6 mm yr−1 that is an order of magnitude higher than previously obtained rates averaged over longer time periods. Automated extraction and correlation of coastal terrace remnants from digital topography uncovers a symmetrical uplift with diameter of more than 1,000 km. The wavelength and relatively short timescale of the uplift suggest that it is associated with a mantle process, possibly convective upwelling, and that the topography may be modulated by rapid short-lived pulses of mantle-derived uplift. Our study shows that stable continental regions far from the effects of glacial rebound may experience rapid vertical displacements of several millimetres per year