Monomeric ephrinB2 binding induces allosteric changes in Nipah virus G that precede its full activation.

Nipah virus is an emergent paramyxovirus that causes deadly encephalitis and respiratory infections in humans. Two glycoproteins coordinate the infection of host cells, an attachment protein (G), which binds to cell surface receptors, and a fusion (F) protein, which carries out the process of virus-cell membrane fusion. The G protein binds to ephrin B2/3 receptors, inducing G conformational changes that trigger F protein refolding. Using an optical approach based on second harmonic generation, we show that monomeric and dimeric receptors activate distinct conformational changes in G. The monomeric receptor-induced changes are not detected by conformation-sensitive monoclonal antibodies or through electron microscopy analysis of G:ephrinB2 complexes. However, hydrogen/deuterium exchange experiments confirm the second harmonic generation observations and reveal allosteric changes in the G receptor binding and F-activating stalk domains, providing insights into the pathway of receptor-activated virus entry.Nipah virus causes encephalitis in humans. Here the authors use a multidisciplinary approach to study the binding of the viral attachment protein G to its host receptor ephrinB2 and show that monomeric and dimeric receptors activate distinct conformational changes in G and discuss implications for receptor-activated virus entry

A unified mechanism for intron and exon definition and back-splicing.

The molecular mechanisms of exon definition and back-splicing are fundamental unanswered questions in pre-mRNA splicing. Here we report cryo-electron microscopy structures of the yeast spliceosomal E complex assembled on introns, providing a view of the earliest event in the splicing cycle that commits pre-mRNAs to splicing. The E complex architecture suggests that the same spliceosome can assemble across an exon, and that it either remodels to span an intron for canonical linear splicing (typically on short exons) or catalyses back-splicing to generate circular RNA (on long exons). The model is supported by our experiments, which show that an E complex assembled on the middle exon of yeast EFM5 or HMRA1 can be chased into circular RNA when the exon is sufficiently long. This simple model unifies intron definition, exon definition, and back-splicing through the same spliceosome in all eukaryotes and should inspire experiments in many other systems to understand the mechanism and regulation of these processes

Author Correction: CryoEM structure of Saccharomyces cerevisiae U1 snRNP offers insight into alternative splicing.

The originally published version of this Article contained several errors in Figure 2, panel a: the basepair register in SL3-4 of yeast U1 snRNA was depicted incorrectly; the basepair for A287-U295 in yeast U1 snRNA was erroneously present; basepairs for U84-G119, G309-U532, A288-U295 and U289-A294 in yeast U1 snRNA were missing; the bulging nucleotide in SL3 of human U1 snRNA was depicted as G instead of C; and the dashed boxes defining the 5' ss binding site and Sm site in both human and yeast snRNAs were not drawn accurately. These have now been corrected in both the PDF and HTML versions of the Article

Current situation and related factors of occupational stress of employees of a petrochemical enterprise

BackgroundOccupational stress has become one of the main factors affecting people's physical and mental health, and there are many sources of occupational stress in petrochemical enterprises. ObjectiveTo evaluate the current situation of occupational stress and its related factors among employees in a petrochemical enterprise, and to provide a scientific basis for reduing the risk of occupational stress among employees in petrochemical enterprises. MethodsIn June 2022, a cross-sectional questionnaire survey was conducted in a petrochemical enterprise in Hainan, including a general information questionnaire for basic information, the Effort-Reward Imbalance (ERI) for occupational stress, and the Pittsburgh Sleep Quality Index (PSQI) for sleep quality. Chi-square test was used to compare differences in positive occupational stress by demographic characteristics, occupational characteristics, behavior, and occupational disease hazards. Logistic regression was employed to evaluate factors associated with occupational stress. ResultsOf the 1129 questionnaire distributed, a total of 999 valid questionnaire were returned,with a valid recovery rate of 88.5%. The positive rate of occupational stress among employees in the petrochemical enterprise was 29.5%. There were statistically significant differences in the positive rate of occupational stress among the employees grouped by gender, age, marital status, body mass index (BMI), monthly income, length of service, smoking, weekly working hours, type of work, working mode, sleep quality, noise exposure, and high temperature exposure (P<0.05). In terms of positive occupational stress among subcategories: workers being male (vs. female), working ＞40 h per week (vs. ≤40 h per week), regular day shift (vs. shift work), smoking (vs. not smoking), with exposure to noise and heat (vs. without such exposure), and having poor sleep quality (vs. good sleep quality) reported higher positive occupational stress rates (P＜0.05). The results of pairwise comparison showed that the positive rate of occupational stress in divorced (50.0%) or married (32.0%) workers was higher than that in single (27.1%) workers, and higher in operation workers (30.6%) than in other types of work (20.5%) (P＜0.05). The trend chi-square results showed that the positive rate of occupational stress increased linearly with the increase of age, length of service, BMI, or monthly income (P＜0.05). The results of logistic regression analysis after adjustment showed that workers who worked ＞40 h a week had a higher risk of occupational stress than those who worked ≤40 h a week, and the OR (95%CI) was 1.909 (1.135, 3.211); the workers of other types of work had a lower risk of reporting occupational stress than operation workers, and the OR (95%CI) was 0.513 (0.272, 0.968); the workers with noise exposure had a higher risk of occupational stress than the workers without, and the OR (95%CI) was 2.457 (1.070, 5.642). ConclusionThe positive rate of occupational stress among employees in this petrochemical enterprise is high. Among them, operators, working hours per week＞40 h, and noise exposure may increase the incidence of occupational stress. The enterprise should actively take measures to reduce the occurrence of occupational stress among employees

Protein target highlights in CASP15: Analysis of models by structure providers

We present an in-depth analysis of selected CASP15 targets, focusing on their biological and functional significance. The authors of the structures identify and discuss key protein features and evaluate how effectively these aspects were captured in the submitted predictions. While the overall ability to predict three-dimensional protein structures continues to impress, reproducing uncommon features not previously observed in experimental structures is still a challenge. Furthermore, instances with conformational flexibility and large multimeric complexes highlight the need for novel scoring strategies to better emphasize biologically relevant structural regions. Looking ahead, closer integration of computational and experimental techniques will play a key role in determining the next challenges to be unraveled in the field of structural molecular biology

Structure, dynamics and assembly of the ankyrin complex on human red blood cell membrane

The cytoskeleton of a red blood cell (RBC) is anchored to the cell membrane by the ankyrin complex. This complex is assembled during RBC genesis and comprises primarily band 3, protein 4.2 and ankyrin, whose mutations contribute to numerous human inherited diseases. High-resolution structures of the ankyrin complex have been long sought-after to understand its assembly and disease-causing mutations. Here, we analyzed native complexes on the human RBC membrane by stepwise fractionation. Cryo-electron microscopy structures of nine band-3-associated complexes reveal that protein 4.2 stabilizes the cytoplasmic domain of band 3 dimer. In turn, the superhelix-shaped ankyrin binds to this protein 4.2 via ankyrin repeats (ARs) 6-13 and to another band 3 dimer via ARs 17-20, bridging two band 3 dimers in the ankyrin complex. Integration of these structures with both prior data and our biochemical data supports a model of ankyrin complex assembly during erythropoiesis and identifies interactions essential for the mechanical stability of RBC

Experimental Investigation on Asphalt Binders Ageing Behavior and Rejuvenating Feasibility in Multicycle Repeated Ageing and Recycling

Multicycle repeated utilization of reclaimed asphalt pavement (RAP) is a quite recent development of sustainable pavement materials technology. To investigate ageing rules and recycling possibility of asphalt binders in repeatedly used asphalt mixture, virgin asphalt AH-70 samples were heated by the rolling thin film oven test (RTFOT) at 163°C, respectively, for 40, 85, 180, 240, and 300 minutes to simulate different ageing degrees, and then the aged ones were rejuvenated by adding a self-made rejuvenator. This ageing and recycling process was repeated altogether for 5 cycles to simulate repeated use of RAP binders. In repeated recycling, rejuvenator contents for different cycle numbers or ageing durations were not the same, and the optimum ones were initially estimated by an empirical formula and finally obtained by comparative tests. Empirical rheological tests and the infrared spectral (IR) analysis were done before and after each cycle of recycling. Results indicate that for impact on deterioration of asphalt binders, ageing time is more important than cycle number. Meanwhile, the asphalt after multicycle repeated ageing and recycling can be restored to the empirical rheological indices level of the virgin asphalt and meet specifications requirements

Structures and comparison of endogenous 2-oxoglutarate and pyruvate dehydrogenase complexes from bovine kidney

The α-keto acid dehydrogenase complex family catalyzes the essential oxidative decarboxylation of α-keto acids to yield acyl-CoA and NADH. Despite performing the same overarching reaction, members of the family have different component structures and structural organization between each other and across phylogenetic species. While native structures of α-keto acid dehydrogenase complexes from bacteria and fungi became available recently, the atomic structure and organization of their mammalian counterparts in native states remain unknown. Here, we report the cryo-electron microscopy structures of the endogenous cubic 2-oxoglutarate dehydrogenase complex (OGDC) and icosahedral pyruvate dehydrogenase complex (PDC) cores from bovine kidney determined at resolutions of 3.5 Å and 3.8 Å, respectively. The structures of multiple proteins were reconstructed from a single lysate sample, allowing direct structural comparison without the concerns of differences arising from sample preparation and structure determination. Although native and recombinant E2 core scaffold structures are similar, the native structures are decorated with their peripheral E1 and E3 subunits. Asymmetric sub-particle reconstructions support heterogeneity in the arrangements of these peripheral subunits. In addition, despite sharing a similar monomeric fold, OGDC and PDC E2 cores have distinct interdomain and intertrimer interactions, which suggests a means of modulating self-assembly to mitigate heterologous binding between mismatched E2 species. The lipoyl moiety lies near a mobile gatekeeper within the interdomain active site of OGDC E2 and PDC E2. Analysis of the twofold related intertrimer interface identified secondary structural differences and chemical interactions between icosahedral and cubic geometries of the core. Taken together, our study provides a direct structural comparison of OGDC and PDC from the same source and offers new insights into determinants of interdomain interactions and of architecture diversity among α-keto acid dehydrogenase complexes

Impact of Land Cover Changes on Soil Type Mapping in Plain Areas: Evidence from Tongzhou District of Beijing, China

The flat terrain in the plain areas of Beijing, China makes the land easily accessible for cultivation and farming, providing vast opportunities for agricultural development. Meanwhile, these areas are also crucial for urban construction and economic growth. Soil type mapping plays a key role in understanding soil characteristics and guiding land management practices. However, accurately mapping soil types in plain regions can be challenging due to their low spatial variability and diverse land use types. Although land cover changes due to phenomena such as urbanization, agricultural expansion, and conversion of natural vegetation can significantly affect soil properties and distribution patterns, their impacts on soil type mapping remain unclear. This study investigated the impacts of land cover changes in plain areas on the accuracy of soil type mapping, hoping to provide effective assistance for soil type mapping in plain areas by analyzing their coupling relationship. Focusing on the 20 year land cover changes in Tongzhou District, this study utilizes a unified approach that combines expert knowledge, mixed sampling methods, and RF mapping techniques, while incorporating environmental covariates that have minimal period influence and synergistically using NDVI and land cover data from the same year. Transition matrices are used to reveal land cover changes, confusion matrices, and their derived indicators to analyze changes in soil type mapping accuracy, and coupling analysis is conducted between soil type change areas and land cover change areas. The results show that Tongzhou District has experienced rapid development over the past 20 years, with the area of construction land nearly doubling. Additionally, 29% of arable land has been converted into construction land, resulting in an increase in the accuracy of the soil map from 58.99% to 66.91% over the 20 year period. The soil type change area during this period accounts for 16.5% of the total area, with 51.9% of the changed areas overlapping with land cover change areas. These overlapping regions are predominantly influenced by human activities. In terms of cultivated land types in the study area, the quantity of arable land has decreased by approximately 29% over 20 years, while the proportion of Sandy loam calcareous fluvo-aquic soil and Light loam calcareous fluvo-aquic soil, which constitute nearly half of the soil type, has increased. These data demonstrate the coupling relationship between land cover changes and soil type variations. It is evident that improving the extent of land use in plain areas enhances the credibility of soil type mapping. Meanwhile, human activities impact land cover, which, in turn, affects and reflects changes in the soil type

Atomic structures of anthrax toxin protective antigen channels bound to partially unfolded lethal and edema factors.

Following assembly, the anthrax protective antigen (PA) forms an oligomeric translocon that unfolds and translocates either its lethal factor (LF) or edema factor (EF) into the host cell. Here, we report the cryo-EM structures of heptameric PA channels with partially unfolded LF and EF at 4.6 and 3.1-Å resolution, respectively. The first α helix and β strand of LF and EF unfold and dock into a deep amphipathic cleft, called the α clamp, which resides at the interface of two PA monomers. The α-clamp-helix interactions exhibit structural plasticity when comparing the structures of lethal and edema toxins. EF undergoes a largescale conformational rearrangement when forming the complex with the channel. A critical loop in the PA binding interface is displaced for about 4 Å, leading to the weakening of the binding interface prior to translocation. These structures provide key insights into the molecular mechanisms of translocation-coupled protein unfolding and translocation