Cooperativity and Frustration in Protein-Mediated Parallel Actin Bundles

We examine the mechanism of bundling of cytoskeletal actin filaments by two representative bundling proteins, fascin and espin. Small-angle X-ray studies show that increased binding from linkers drives a systematic \textit{overtwist} of actin filaments from their native state, which occurs in a linker-dependent fashion. Fascin bundles actin into a continuous spectrum of intermediate twist states, while espin only allows for untwisted actin filaments and fully-overtwisted bundles. Based on a coarse-grained, statistical model of protein binding, we show that the interplay between binding geometry and the intrinsic \textit{flexibility} of linkers mediates cooperative binding in the bundle. We attribute the respective continuous/discontinous bundling mechanisms of fascin/espin to differences in the stiffness of linker bonds themselves.Comment: 5 pages, 3 figures, figure file has been corrected in v

The AP-2 clathrin adaptor mediates endocytosis of an inhibitory killer cell Ig-like receptor in human NK cells

Copyright © 2014 by The American Association of Immunologists, Inc. All rights reserved. Stable surface expression of human inhibitory killer cell Ig-like receptors (KIRs) is critical for controlling NK cell function and maintaining NK cell tolerance toward normal MHC class I+ cells. Our recent experiments, however, have found that Ab-bound KIR3DL1 (3DL1) readily leaves the cell surface and undergoes endocytosis to early/recycling endosomes and subsequently to late endosomes. We found that 3DL1 internalization is at least partially mediated by an interaction between the μ2 subunit of the AP-2 clathrin adaptor complex and ITIM tyrosine residues in the cytoplasmic domain of 3DL1. Disruption of the 3DL1/μ2 interaction, either by mutation of the ITIM tyrosines in 3DL1 or mutation of μ2, significantly diminished endocytosis and increased surface expression of 3DL1 in human primary NK cells and cell lines. Furthermore, we found that the 3DL1/AP-2 interaction is diminished upon Ab engagement with the receptor, as compared with untreated cells. Thus, we have identified AP-2-mediated endocytosis as a mechanism regulating the surface levels of inhibitory KIRs through their ITIM domains. Based on our results, we propose a model in which nonengaged KIRs are internalized by this mechanism, whereas engagement with MHC class I ligand would diminish AP-2 binding, thereby prolonging stable receptor surface expression and promoting inhibitory function. Furthermore, this ITIM-mediated mechanism may similarly regulate the surface expression of other inhibitory immune receptors

Clinical-dosimetric analysis of measures of dysphagia including gastrostomy-tube dependence among head and neck cancer patients treated definitively by intensity-modulated radiotherapy with concurrent chemotherapy

<p>Abstract</p> <p>Purpose</p> <p>To investigate the association between dose to various anatomical structures and dysphagia among patients with head and neck cancer treated by definitive intensity-modulated radiotherapy (IMRT) and concurrent chemotherapy.</p> <p>Methods and materials</p> <p>Thirty-nine patients with squamous cancer of the head and neck were treated by definitive concurrent chemotherapy and IMRT to a median dose of 70 Gy (range, 68 to 72). In each patient, a gastrostomy tube (GT) was prophylacticly placed prior to starting treatment. Prolonged GT dependence was defined as exceeding the median GT duration of 192 days. Dysphagia was scored using standardized quality-of-life instruments. Dose-volume histogram (DVH) data incorporating the superior/middle pharyngeal constrictors (SMPC), inferior pharyngeal constrictor (IPC), cricoid pharyngeal inlet (CPI), and cervical esophagus (CE) were analyzed in relation to prolonged GT dependence, dysphagia, and weight loss.</p> <p>Results</p> <p>At 3 months and 6 months after treatment, 87% and 44% of patients, respectively, were GT dependent. Spearman's ρ analysis identified statistical correlations (p < 0.05) between prolonged GT dependence or high grade dysphagia with IPC V65, IPC V60, IPC Dmean, and CPI Dmax. Logistic regression model showed that IPC V65 > 30%, IPC V60 > 60%, IPC Dmean > 60 Gy, and CPI Dmax > 62 Gy predicted for greater than 50% probability of prolonged GT dependence.</p> <p>Conclusion</p> <p>Our analysis suggests that adhering to the following parameters may decrease the risk of prolonged GT dependence and dysphagia: IPC V65 < 15%, IPC V60 < 40%, IPC Dmean < 55 Gy, and CPI Dmax < 60 Gy.</p

The future of evapotranspiration : global requirements for ecosystem functioning, carbon and climate feedbacks, agricultural management, and water resources

The fate of the terrestrial biosphere is highly uncertain given recent and projected changes in climate. This is especially acute for impacts associated with changes in drought frequency and intensity on the distribution and timing of water availability. The development of effective adaptation strategies for these emerging threats to food and water security are compromised by limitations in our understanding of how natural and managed ecosystems are responding to changing hydrological and climatological regimes. This information gap is exacerbated by insufficient monitoring capabilities from local to global scales. Here, we describe how evapotranspiration (ET) represents the key variable in linking ecosystem functioning, carbon and climate feedbacks, agricultural management, and water resources, and highlight both the outstanding science and applications questions and the actions, especially from a space-based perspective, necessary to advance them

The Hepatitis E Virus Polyproline Region Is Involved in Viral Adaptation

Genomes of hepatitis E virus (HEV), rubivirus and cutthroat virus (CTV) contain a region of high proline density and low amino acid (aa) complexity, named the polyproline region (PPR). In HEV genotypes 1, 3 and 4, it is the only region within the non-structural open reading frame (ORF1) with positive selection (4–10 codons with dN/dS>1). This region has the highest density of sites with homoplasy values >0.5. Genotypes 3 and 4 show ∼3-fold increase in homoplastic density (HD) in the PPR compared to any other region in ORF1, genotype 1 does not exhibit significant HD (p<0.0001). PPR sequence divergence was found to be 2-fold greater for HEV genotypes 3 and 4 than for genotype 1. The data suggest the PPR plays an important role in host-range adaptation. Although the PPR appears to be hypervariable and homoplastic, it retains as much phylogenetic signal as any other similar sized region in the ORF1, indicating that convergent evolution operates within the major HEV phylogenetic lineages. Analyses of sequence-based secondary structure and the tertiary structure identify PPR as an intrinsically disordered region (IDR), implicating its role in regulation of replication. The identified propensity for the disorder-to-order state transitions indicates the PPR is involved in protein-protein interactions. Furthermore, the PPR of all four HEV genotypes contains seven putative linear binding motifs for ligands involved in the regulation of a wide number of cellular signaling processes. Structure-based analysis of possible molecular functions of these motifs showed the PPR is prone to bind a wide variety of ligands. Collectively, these data suggest a role for the PPR in HEV adaptation. Particularly as an IDR, the PPR likely contributes to fine tuning of viral replication through protein-protein interactions and should be considered as a target for development of novel anti-viral drugs

Digitise This! A Quick and Easy Remote Sensing Method to Monitor the Daily Extent of Dredge Plumes

Technological advancements in remote sensing and GIS have improved natural resource managers’ abilities to monitor large-scale disturbances. In a time where many processes are heading towards automation, this study has regressed to simple techniques to bridge a gap found in the advancement of technology. The near-daily monitoring of dredge plume extent is common practice using Moderate Resolution Imaging Spectroradiometer (MODIS) imagery and associated algorithms to predict the total suspended solids (TSS) concentration in the surface waters originating from floods and dredge plumes. Unfortunately, these methods cannot determine the difference between dredge plume and benthic features in shallow, clear water. This case study at Barrow Island, Western Australia, uses hand digitising to demonstrate the ability of human interpretation to determine this difference with a level of confidence and compares the method to contemporary TSS methods. Hand digitising was quick, cheap and required very little training of staff to complete. Results of ANOSIM R statistics show remote sensing derived TSS provided similar spatial results if they were thresholded to at least 3 mg L-1. However, remote sensing derived TSS consistently provided false-positive readings of shallow benthic features as Plume with a threshold up to TSS of 6 mg L-1, and began providing false-negatives (excluding actual plume) at a threshold as low as 4 mg L-1. Semi-automated processes that estimate plume concentration and distinguish between plumes and shallow benthic features without the arbitrary nature of human interpretation would be preferred as a plume monitoring method. However, at this stage, the hand digitising method is very useful and is more accurate at determining plume boundaries over shallow benthic features and is accessible to all levels of management with basic training