There are currently limited Food and Drug Administration (FDA)-approved drugs and vaccines for the treatment or prevention of Coronavirus Disease 2019 (COVID-19). Enhanced understanding of Severe ...Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection and pathogenesis is critical for the development of therapeutics. To provide insight into viral replication, cell tropism, and host-viral interactions of SARS-CoV-2, we performed single-cell (sc) RNA sequencing (RNA-seq) of experimentally infected human bronchial epithelial cells (HBECs) in air-liquid interface (ALI) cultures over a time course. This revealed novel polyadenylated viral transcripts and highlighted ciliated cells as a major target at the onset of infection, which we confirmed by electron and immunofluorescence microscopy. Over the course of infection, the cell tropism of SARS-CoV-2 expands to other epithelial cell types including basal and club cells. Infection induces cell-intrinsic expression of type I and type III interferons (IFNs) and interleukin (IL)-6 but not IL-1. This results in expression of interferon-stimulated genes (ISGs) in both infected and bystander cells. This provides a detailed characterization of genes, cell types, and cell state changes associated with SARS-CoV-2 infection in the human airway.
In most Mediterranean climate (MedClim) regions around the world, global climate models (GCMs) consistently project drier futures. In California, however, projections of changes in annual ...precipitation are inconsistent. Analysis of daily precipitation in 30 GCMs reveals patterns in projected hydrometeorology over each of the five MedClm regions globally and helps disentangle their causes. MedClim regions, except California, are expected to dry via decreased frequency of winter precipitation. Frequencies of extreme precipitation, however, are projected to increase over the two MedClim regions of the Northern Hemisphere where projected warming is strongest. The increase in heavy and extreme precipitation is particularly robust over California, where it is only partially offset by projected decreases in low-medium intensity precipitation. Over the Mediterranean Basin, however, losses from decreasing frequency of low-medium-intensity precipitation are projected to dominate gains from intensifying projected extreme precipitation. MedClim regions are projected to become more sub-tropical, i.e. made dryer via pole-ward expanding subtropical subsidence. California's more nuanced hydrological future reflects a precarious balance between the expanding subtropical high from the south and the south-eastward extending Aleutian low from the north-west. These dynamical mechanisms and thermodynamic moistening of the warming atmosphere result in increased horizontal water vapor transport, bolstering extreme precipitation events.
When applied to remove climate model biases in precipitation, quantile mapping can in some settings modify the simulated difference in mean precipitation between two eras. This has important ...implications when the precipitation is used to drive an impacts model that is sensitive to changes in precipitation. The tendency of quantile mapping to alter model-predicted changes is demonstrated using synthetic precipitation distributions and elucidated with a simple theoretical analysis, which shows that the alteration of model-predicted changes can be controlled by the ratio of model to observed variance. To further evaluate the effects of quantile mapping in a more realistic setting, we use daily precipitation output from 11 atmospheric general circulation models (AGCMs), forced by observed sea surface temperatures, over the conterminous United States to compare precipitation differences before and after quantile mapping bias correction. The effectiveness of the bias correction is not assessed, only its effect on precipitation differences. The change in seasonal mean (winter, DJF, and summer, JJA) precipitation between two historical periods is compared to examine whether the bias correction tends to amplify or diminish an AGCM's simulated precipitation change. In some cases the trend modification can be as large as the original simulated change, though the areas where this occurs varies among AGCMs so the ensemble median shows smaller trend modification. Results show that quantile mapping improves the correspondence with observed changes in some locations and degrades it in others. While not representative of a future where natural precipitation variability is much smaller than that due to external forcing, these results suggest that at least for the next several decades the influence of quantile mapping on seasonal precipitation trends does not systematically degrade projected differences.
Using small molecules that trap translation factors within translating ribosomes, Gurzeler et al.1 and Oltion et al.2 identify a new branch of the ribosome-associated quality-control (RQC) pathway. ...This mode of translation regulation expands the number of mechanistically distinct RQC pathways.
Using small molecules that trap translation factors within translating ribosomes, Gurzeler et al.1 and Oltion et al.2 identify a new branch of the ribosome-associated quality-control (RQC) pathway. This mode of translation regulation expands the number of mechanistically distinct RQC pathways.
The modular SCF (Skp1, cullin, and F box) ubiquitin ligases feature a large family of F box protein substrate receptors that enable recognition of diverse targets. However, how the repertoire of SCF ...complexes is sustained remains unclear. Real-time measurements of formation and disassembly indicate that SCFFbxw7 is extraordinarily stable, but, in the Nedd8-deconjugated state, the cullin-binding protein Cand1 augments its dissociation by one-million-fold. Binding and ubiquitylation assays show that Cand1 is a protein exchange factor that accelerates the rate at which Cul1-Rbx1 equilibrates with multiple F box protein-Skp1 modules. Depletion of Cand1 from cells impedes recruitment of new F box proteins to pre-existing Cul1 and profoundly alters the cellular landscape of SCF complexes. We suggest that catalyzed protein exchange may be a general feature of dynamic macromolecular machines and propose a hypothesis for how substrates, Nedd8, and Cand1 collaborate to regulate the cellular repertoire of SCF complexes.
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► Cand1 modulates the repertoire of SCF ubiquitin ligase complexes in cells ► Cand1 functions as an F box protein exchange factor ► FRET assay reveals real-time dynamics of SCFFbxw7 assembly and disassembly ► Cand1 accelerates the dissociation rate of SCFFbxw7 one-million-fold
Cand1 accelerates the dissociation of F box proteins from SCF ubiquitin ligases by one-million-fold. It thereby operates as a protein exchange factor to shape the cellular repertoire of SCF complexes.
Floodplain buyouts are an increasingly common policy for mitigating flood risk. Recent research has explored the costs of buyouts and their impacts on municipal finance and tax base. However, little ...work has explored the effects of buyouts on surrounding residential land values, an aspect that could contribute to the extensive literature on the land value impacts of urban land uses, including open space and ecological restoration. This study evaluates the residential land value impacts of buyouts in Mecklenburg County, North Carolina (USA), an area with extensive, municipally- and federally-funded buyouts (n = 348). Using a quasi-experimental research design that matches treatment parcels (near buyouts) to control parcels to isolate the causal impacts of buyouts, we find that buyouts are responsible for weak increases (p<0.1) in the area-normalized sales values ($/ft2) of neighboring (within 0.15 km) single-family residential properties. We additionally find positive – but also weak – sales value relationships with (1) buyouts that have programmed post-buyout land uses, such as parks or other recreation areas (USD$25.46/ft2 $274.05/m2 when 0.15–0.2 km from buyout; p<0.05), and (2) the average age of proximal buyouts ($0.34/ft2 $3.66/m2 within 0.1–0.15 km; $0.30/ft2 $3.23/m2 within 0.15–0.20 km; both p<0.01). Our results suggest that post-buyout land uses may have impacts on local tax base and should play a larger part in municipal buyout decisions.
•We isolate floodplain buyouts' causal impacts using treatment-control parcel matching.•Buyouts weakly increase sales values ($/m2) of nearby (<0.15 km) residential parcels.•Decisions on post-buyout land uses could impact local tax base.
Polymerization of actin filaments directed by the actin-related protein (Arp)2/3 complex supports many types of cellular movements. However, questions remain regarding the relative contributions of ...Arp2/3 complex versus other mechanisms of actin filament nucleation to processes such as path finding by neuronal growth cones; this is because of the lack of simple methods to inhibit Arp2/3 complex reversibly in living cells. Here we describe two classes of small molecules that bind to different sites on the Arp2/3 complex and inhibit its ability to nucleate actin filaments. CK-0944636 binds between Arp2 and Arp3, where it appears to block movement of Arp2 and Arp3 into their active conformation. CK-0993548 inserts into the hydrophobic core of Arp3 and alters its conformation. Both classes of compounds inhibit formation of actin filament comet tails by Listeria and podosomes by monocytes. Two inhibitors with different mechanisms of action provide a powerful approach for studying the Arp2/3 complex in living cells.
Global climate model (GCM) output typically needs to be bias corrected before it can be used for climate change impact studies. Three existing bias correction methods, and a new one developed here, ...are applied to daily maximum temperature and precipitation from 21 GCMs to investigate how different methods alter the climate change signal of the GCM. The quantile mapping (QM) and cumulative distribution function transform (CDF-t) bias correction methods can significantly alter the GCM’s mean climate change signal, with differences of up to 2°C and 30% points for monthly mean temperature and precipitation, respectively. Equidistant quantile matching (EDCDFm) bias correction preserves GCM changes in mean daily maximum temperature but not precipitation. An extension to EDCDFm termed PresRat is introduced, which generally preserves the GCM changes in mean precipitation. Another problem is that GCMs can have difficulty simulating variance as a function of frequency. To address this, a frequency-dependent bias correction method is introduced that is twice as effective as standard bias correction in reducing errors in the models’ simulation of variance as a function of frequency, and it does so without making any locations worse, unlike standard bias correction. Last, a preconditioning technique is introduced that improves the simulation of the annual cycle while still allowing the bias correction to take account of an entire season’s values at once.
A number of clinically validated drugs have been developed by repurposing the CUL4-DDB1-CRBN-RBX1 (CRL4CRBN) E3 ubiquitin ligase complex with molecular glue degraders to eliminate disease-driving ...proteins. Here, we present the identification of a first-in-class GSPT1-selective cereblon E3 ligase modulator, CC-90009. Biochemical, structural, and molecular characterization demonstrates that CC-90009 coopts the CRL4CRBN to selectively target GSPT1 for ubiquitination and proteasomal degradation. Depletion of GSPT1 by CC-90009 rapidly induces acute myeloid leukemia (AML) apoptosis, reducing leukemia engraftment and leukemia stem cells (LSCs) in large-scale primary patient xenografting of 35 independent AML samples, including those with adverse risk features. Using a genome-wide CRISPR-Cas9 screen for effectors of CC-90009 response, we uncovered the ILF2 and ILF3 heterodimeric complex as a novel regulator of cereblon expression. Knockout of ILF2/ILF3 decreases the production of full-length cereblon protein via modulating CRBN messenger RNA alternative splicing, leading to diminished response to CC-90009. The screen also revealed that the mTOR signaling and the integrated stress response specifically regulate the response to CC-90009 in contrast to other cereblon modulators. Hyperactivation of the mTOR pathway by inactivation of TSC1 and TSC2 protected against the growth inhibitory effect of CC-90009 by reducing CC-90009-induced binding of GSPT1 to cereblon and subsequent GSPT1 degradation. On the other hand, GSPT1 degradation promoted the activation of the GCN1/GCN2/ATF4 pathway and subsequent apoptosis in AML cells. Collectively, CC-90009 activity is mediated by multiple layers of signaling networks and pathways within AML blasts and LSCs, whose elucidation gives insight into further assessment of CC-90009s clinical utility. These trials were registered at www.clinicaltrials.gov as #NCT02848001 and #NCT04336982).
An interaction that guides cell fate
Notch signaling is important in cell fate determination in mammals. Signaling is initiated when the extracellular domain of the transmembrane Notch protein on one ...cell binds to a surface ligand on another cell. Luca
et al.
report the crystal structure of the interacting regions of Notch and the Delta-like ligand DLL-4. The Notch protein is modified by O-linked glycan addition, and this is required for signaling. The structure shows two interaction interfaces. A glycan anchors the less conserved interface, which potentially provides a flexible way of regulating Notch interactions during development.
Science
, this issue p.
847
O-linked glycosylation regulates a receptor-ligand interaction that is important in cell fate determination.
Notch receptors guide mammalian cell fate decisions by engaging the proteins Jagged and Delta-like (DLL). The 2.3 angstrom resolution crystal structure of the interacting regions of the Notch1-DLL4 complex reveals a two-site, antiparallel binding orientation assisted by Notch1 O-linked glycosylation. Notch1 epidermal growth factor–like repeats 11 and 12 interact with the DLL4 Delta/Serrate/Lag-2 (DSL) domain and module at the N-terminus of Notch ligands (MNNL) domains, respectively. Threonine and serine residues on Notch1 are functionalized with O-fucose and O-glucose, which act as surrogate amino acids by making specific, and essential, contacts to residues on DLL4. The elucidation of a direct chemical role for O-glycans in Notch1 ligand engagement demonstrates how, by relying on posttranslational modifications of their ligand binding sites, Notch proteins have linked their functional capacity to developmentally regulated biosynthetic pathways.
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