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Systemic capillary leak syndrome (SCLS) is a rare life-threatening disorder due to profound vascular leak. The trigger and the cause of the disease are currently unknown and there is no specific treatment. Here, we identified a rare heterozygous splice-site variant in the TLN1 gene in a familial SCLS case, suggestive of autosomal dominant inheritance with incomplete penetrance. Talin1 has a key role in cell adhesion by activating and linking integrins to the actin cytoskeleton. This variant causes in-frame skipping of exon 54 and is predicted to affect talin's C-terminal actin-binding site (ABS3). Modeling the SCLS-TLN1 variant in TLN1-heterozygous endothelial cells (ECs) disturbed the endothelial barrier function. Similarly, mimicking the predicted actin-binding disruption in TLN1-heterozygous ECs resulted in disorganized endothelial adherens junctions. Mechanistically, we established that the SCLS-TLN1 variant, through the disruption of talin's ABS3, sequestrates talin's interacting partner, vinculin, at cell-extracellular matrix adhesions, leading to destabilization of the endothelial barrier. We propose that pathogenic variants in TLN1 underlie SCLS, providing insight into the molecular mechanism of the disease that can be explored for future therapeutic interventions.

Misprocessing of amyloid precursor protein (APP) is one of the major causes of Alzheimer's disease. APP comprises a large extracellular region, a single transmembrane helix and a short cytoplasmic tail containing an NPxY motif (normally referred to as the YENPTY motif). Talins are synaptic scaffold proteins that connect the cytoskeletal machinery to the plasma membrane via binding NPxY motifs in the cytoplasmic tail of integrins. Here, we report the crystal structure of an APP/talin1 complex identifying a new way to couple the cytoskeletal machinery to synaptic sites through APP. Proximity ligation assay (PLA) confirmed the close proximity of talin1 and APP in primary neurons, and talin1 depletion had a dramatic effect on APP processing in cells. Structural modelling reveals APP might form an extracellular meshwork that mechanically couples the cytoskeletons of the pre- and post-synaptic compartments. We propose APP processing represents a mechanical signalling pathway whereby under tension, the cleavage sites in APP have varying accessibility to cleavage by secretases. This leads us to propose a new hypothesis for Alzheimer's, where misregulated APP dynamics result in loss of the mechanical integrity of the synapse, corruption and loss of mechanical binary data, and excessive generation of toxic plaque-forming Aβ42 peptide.

We previously identified talin rod domain-containing protein 1 (TLNRD1) as a potent actin-bundling protein in vitro. Here, we report that TLNRD1 is expressed in the vasculature in vivo. Its depletion leads to vascular abnormalities in vivo and modulation of endothelial cell monolayer integrity in vitro. We demonstrate that TLNRD1 is a component of the cerebral cavernous malformations (CCM) complex through its direct interaction with CCM2, which is mediated by a hydrophobic C-terminal helix in CCM2 that attaches to a hydrophobic groove on the four-helix domain of TLNRD1. Disruption of this binding interface leads to CCM2 and TLNRD1 accumulation in the nucleus and actin fibers. Our findings indicate that CCM2 controls TLNRD1 localization to the cytoplasm and inhibits its actin-bundling activity and that the CCM2-TLNRD1 interaction impacts endothelial actin stress fiber and focal adhesion formation. Based on these results, we propose a new pathway by which the CCM complex modulates the actin cytoskeleton and vascular integrity., (© 2024 Ball et al.)

Vinculin binds to specific sites of mechanically unfolded talin rod domains to reinforce the coupling of the cell's exterior to its force generation machinery. Force-dependent vinculin-talin complexation and dissociation was previously observed as contraction or extension of the unfolded talin domains respectively using magnetic tweezers. However, the structural mechanism underlying vinculin recognition of unfolded vinculin binding sites (VBSs) in talin remains unknown. Using molecular dynamics simulations, we demonstrate that a VBS dynamically refolds under force, and that vinculin can recognize and bind to partially unfolded VBS states. Vinculin binding enables refolding of the mechanically strained VBS and stabilizes its folded α-helical conformation, providing resistance against mechanical stress. Together, these results provide an understanding of a recognition mechanism of proteins unfolded by force and insight into the initial moments of how vinculin binds unfolded talin rod domains during the assembly of this mechanosensing meshwork., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Mykuliak et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Cellular signalling is a complex process and involves cascades of enzymes that, in response to a specific signal, give rise to exact cellular responses. Signalling scaffold proteins organise components of these signalling pathways in space and time to co-ordinate signalling outputs. In this review we introduce a new class of mechanically operated signalling scaffolds that are built into the cytoskeletal architecture of the cell. These proteins contain force-dependent binary switch domains that integrate chemical and mechanical signals to introduce quantised positional changes to ligands and persistent alterations in cytoskeletal architecture providing mechanomemory capabilities. We focus on the concept of spatial organisation, and how the cell organises signalling molecules at the plasma membrane in response to specific signals to create order and distinct signalling outputs. The dynamic positioning of molecules using binary switches adds an additional layer of complexity to the idea of scaffolding. The switches can spatiotemporally organise enzymes and substrates dynamically, with the introduction of ∼50 nm quantised steps in distance between them as the switch patterns change. Together these different types of signalling scaffolds and the proteins engaging them, provide a way for an ordering of molecules that extends beyond current views of the cell., (© 2024 The Author(s).)

Myosin-X (MYO10), a molecular motor localizing to filopodia, is thought to transport various cargo to filopodia tips, modulating filopodia function. However, only a few MYO10 cargoes have been described. Here, using GFP-Trap and BioID approaches combined with mass spectrometry, we identified lamellipodin (RAPH1) as a novel MYO10 cargo. We report that the FERM domain of MYO10 is required for RAPH1 localization and accumulation at filopodia tips. Previous studies have mapped the RAPH1 interaction domain for adhesome components to its talin-binding and Ras-association domains. Surprisingly, we find that the RAPH1 MYO10-binding site is not within these domains. Instead, it comprises a conserved helix located just after the RAPH1 pleckstrin homology domain with previously unknown functions. Functionally, RAPH1 supports MYO10 filopodia formation and stability but is not required to activate integrins at filopodia tips. Taken together, our data indicate a feed-forward mechanism whereby MYO10 filopodia are positively regulated by MYO10-mediated transport of RAPH1 to the filopodium tip., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2023. Published by The Company of Biologists Ltd.)

The popularity of alcohol mixed with energy drinks (AmED) among young adults has spurred studies that focus on its links to aggression and risk-taking behaviors, including risky sex and sexual victimization. However, no studies to date have looked at the relationship between AmED and causes of interpersonal conflict in bars at the event level. The present study evaluated whether AmED use at the time of the bar conflict was associated with greater odds that a bar conflict would be precipitated by sexually related causes. Online survey data, including a description of a recent bar conflict, were collected from a community sample of 175 young adult (97 female) AmED users age 18 to 30 in western New York state. Qualitative findings included the natural categorization of sexually related causes of conflict, consisting of unwanted sexual advances and jealousy, and the prominence of sexual competition in these conflicts. Proportion of AmED use (out of the total quantity of alcoholic drinks) predicted the odds that the bar conflict would have a sexually related cause, above and beyond control variables. How AmED use might be associated with sexual competition and conflict in the bar is discussed.

Auditory detection can improve with practice. These improvements are often assumed to arise from selective attention processes, but longer-term plasticity as a result of training may also play a role. Here, listeners were trained to detect either an 861-Hz or 1058-Hz tone (counterbalanced across participants) presented in noise at SNRs varying from -10 to -24 dB. On the following day, they were tasked with detecting 861-Hz and 1058-Hz tones at an SNR of -21 dB. In between blocks of this active task, EEG was recorded during passive presentation of trained and untrained frequency tones in quiet. Detection accuracy and confidence ratings were higher for trials at listeners' trained, than untrained-frequency (i.e., learning occurred). During passive exposure to sounds, the P2 component of the auditory evoked potential (∼150 - 200 ms post tone onset) was larger in amplitude for the trained compared to the untrained frequency. An analysis of global field power similarly yielded a stronger response for trained tones in the P2 time window. These effects were obtained during passive exposure, suggesting that training induced improvements in detection are not solely related to changes in selective attention. Rather, there may be an important role for changes in the long-term neural representations of sounds., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The HML2 (HERV-K) group constitutes the most recently acquired family of human endogenous retroviruses, with many proviruses less than one million years old. Many maintain intact open reading frames and provirus expression together with HML2 particle formation are observed in early stage human embryo development and are associated with pluripotency as well as inflammatory disease, cancers and HIV-1 infection. Here, we reconstruct the core structural protein (CA) of an HML2 retrovirus, assemble particles in vitro and employ single particle cryogenic electron microscopy (cryo-EM) to determine structures of four classes of CA Fullerene shell assemblies. These icosahedral and capsular assemblies reveal at high-resolution the molecular interactions that allow CA to form both pentamers and hexamers and show how invariant pentamers and structurally plastic hexamers associate to form the unique polyhedral structures found in retroviral cores.

IGF2 mRNA-binding protein 1 (IMP1) is a key regulator of messenger RNA (mRNA) metabolism and transport in organismal development and, in cancer, its mis-regulation is an important component of tumour metastasis. IMP1 function relies on the recognition of a diverse set of mRNA targets that is mediated by the combinatorial action of multiple RNA-binding domains. Here, we dissect the structure and RNA-binding properties of two key RNA-binding domains of IMP1, KH1 and KH2, and we build a kinetic model for the recognition of RNA targets. Our data and model explain how the two domains are organized as an intermolecular pseudo-dimer and that the important role they play in mRNA target recognition is underpinned by the high RNA-binding affinity and fast kinetics of this KH1KH2-RNA recognition unit. Importantly, the high-affinity RNA-binding by KH1KH2 is achieved by an inter-domain coupling 50-fold stronger than that existing in a second pseudo-dimer in the protein, KH3KH4. The presence of this strong coupling supports a role of RNA re-modelling in IMP1 recognition of known cancer targets., (© The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Despite the recent, widespread trend of consuming alcohol mixed with energy drinks (AmEDs) in commercial bar settings, few studies have examined whether this practice exacerbates the risk of experiencing aggression in bars and licensed establishments. Past studies have been limited to between-subjects comparison analyses that are at risk for bias due to selection effects. The present analysis examines whether a sample of individuals who regularly use AmEDs are at elevated risk for experiencing or perpetrating physical aggression in bars when drinking AmEDs versus when they are drinking noncaffeinated alcohol (NCA) use alone. This within-subject analysis controls for any individual differences that may be related to both AmED use and the tendency to engage in aggressive behavior. An online survey was completed by 175 young adults (78 male) who were frequent bar patrons, used AmEDs regularly, and had experienced at least one recent bar conflict incident. Although NCA use was more common than was AmED use, AmED-involved bar aggression was more frequently reported than was aggression that coincided with NCA use only. Additionally, victimization and perpetration of aggression in bar environments were both more common when AmEDs were used than when only NCA was used. Frequency of going to bars was predictive of rates of experiencing bar aggression only when drinking NCA but not when drinking AmEDs. Results suggest that AmED use introduces a unique risk factor into the bar environment that must be considered in future research and in subsequent interventions meant to reduce the incidence of bar aggression. (PsycINFO Database Record (c) 2019 APA, all rights reserved).

The beneficial effects of enriched environments have been established through a long history of research. Enrichment of the living conditions of captive animals in the form of larger cages, sensory stimulating objects, and opportunities for social interaction and physical exercise, has been shown to reduce emotional reactivity, ameliorate abnormal behaviors, and enhance cognitive functioning. Recently, environmental enrichment research has been extended to humans, in part due to growing interest in its potential therapeutic benefits for children with neurodevelopmental disorders (NDDs). This paper reviews the history of enriched environment research and the use of enriched environments as a developmental intervention in studies of both NDD animal models and children. We argue that while environmental enrichment may sometimes benefit children with NDDs, several methodological factors need to be more closely considered before the efficacy of this approach can be adequately evaluated, including: (i) operationally defining and standardizing enriched environment treatments across studies; (ii) use of control groups and better control over potentially confounding variables; and (iii) a comprehensive theoretical framework capable of predicting when and how environmental enrichment will alter the trajectory of NDDs.

Exosomal miRNA transfer is a mechanism for cell-cell communication that is important in the immune response, in the functioning of the nervous system and in cancer. Syncrip/hnRNPQ is a highly conserved RNA-binding protein that mediates the exosomal partition of a set of miRNAs. Here, we report that Syncrip's amino-terminal domain, which was previously thought to mediate protein-protein interactions, is a cryptic, conserved and sequence-specific RNA-binding domain, designated NURR (N-terminal unit for RNA recognition). The NURR domain mediates the specific recognition of a short hEXO sequence defining Syncrip exosomal miRNA targets, and is coupled by a non-canonical structural element to Syncrip's RRM domains to achieve high-affinity miRNA binding. As a consequence, Syncrip-mediated selection of the target miRNAs implies both recognition of the hEXO sequence by the NURR domain and binding of the RRM domains 5' to this sequence. This structural arrangement enables Syncrip-mediated selection of miRNAs with different seed sequences.

The Spumaretrovirinae, or foamy viruses (FVs) are complex retroviruses that infect many species of monkey and ape. Despite little sequence homology, FV and orthoretroviral Gag proteins perform equivalent functions, including genome packaging, virion assembly, trafficking and membrane targeting. However, there is a paucity of structural information for FVs and it is unclear how disparate FV and orthoretroviral Gag molecules share the same function. To probe the functional overlap of FV and orthoretroviral Gag we have determined the structure of a central region of Gag from the Prototype FV (PFV). The structure comprises two all α-helical domains NtDCEN and CtDCEN that although they have no sequence similarity, we show they share the same core fold as the N- (NtDCA) and C-terminal domains (CtDCA) of archetypal orthoretroviral capsid protein (CA). Moreover, structural comparisons with orthoretroviral CA align PFV NtDCEN and CtDCEN with NtDCA and CtDCA respectively. Further in vitro and functional virological assays reveal that residues making inter-domain NtDCEN-CtDCEN interactions are required for PFV capsid assembly and that intact capsid is required for PFV reverse transcription. These data provide the first information that relates the Gag proteins of Spuma and Orthoretrovirinae and suggests a common ancestor for both lineages containing an ancient CA fold., Competing Interests: The authors have declared that no competing interests exist.

Background: HIV-1 capsid influences viral uncoating and nuclear import. Some capsid is detected in the nucleus but it is unclear if it has any function. We reported that the antibiotic Coumermycin-A1 (C-A1) inhibits HIV-1 integration and that a capsid mutation confers resistance to C-A1, suggesting that capsid might affect post-nuclear entry steps., Results: Here we report that C-A1 inhibits HIV-1 integration in a capsid-dependent way. Using molecular docking, we identify an extended binding pocket delimited by two adjacent capsid monomers where C-A1 is predicted to bind. Isothermal titration calorimetry confirmed that C-A1 binds to hexameric capsid. Cyclosporine washout assays in Jurkat CD4+ T cells expressing engineered human TRIMCyp showed that C-A1 causes faster and greater escape from TRIMCyp restriction. Sub-cellular fractionation showed that small amounts of capsid accumulated in the nuclei of infected cells and C-A1 reduced the nuclear capsid. A105S and N74D capsid mutant viruses did not accumulate capsid in the nucleus, irrespective of C-A1 treatment. Depletion of Nup153, a nucleoporin located at the nuclear side of the nuclear pore that binds to HIV-1 capsid, made the virus less susceptible to TRIMCyp restriction, suggesting that Nup153 may help maintain some integrity of the viral core in the nucleus. Furthermore C-A1 increased binding of CPSF6, a nuclear protein, to capsid., Conclusions: Our results indicate that capsid is involved in post-nuclear entry steps preceding integration.

Background: Young adult use of alcohol mixed with caffeinated energy drinks (AmEDs) has been globally linked with increased odds of interpersonal aggression, compared with the use of alcohol alone. However, no prior research has linked these behaviors at the event level in bar drinking situations. The present study assessed whether AmED use is associated with the perpetration of verbal and physical aggression in bar conflicts at the event level., Methods: In Fall 2014, a community sample of 175 young adult AmED users (55% female) completed a web survey describing a recent conflict experienced while drinking in a bar. Use of both AmED and non-AmED alcoholic drinks in the incident were assessed, allowing calculation of our main predictor variable, the proportion of AmEDs consumed (AmED/total drinks consumed). To measure perpetration of aggression, participants reported on the occurrence of 6 verbal and 6 physical acts during the bar conflict incident., Results: Linear regression analyses showed that the proportion of AmEDs consumed predicted scores for perpetration of both verbal aggression (β = 0.16, p < 0.05) and physical aggression (β = 0.19, p < 0.01) after controlling for gender, age, sensation-seeking and aggressive personality traits, aggressive alcohol expectancies, aggressogenic physical and social bar environments, and total number of drinks., Conclusions: Results of this study suggest that in alcohol-related bar conflicts, higher levels of young adult AmED use are associated with higher levels of aggression perpetration than alcohol use alone and that the elevated risk is not attributable to individual differences between AmED users and nonusers or to contextual differences in bar drinking settings. While future research is needed to identify motivations, dosages, and sequencing issues associated with AmED use, these beverages should be considered a potential risk factor in the escalation of aggressive bar conflicts., (Copyright © 2016 by the Research Society on Alcoholism.)