bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cryo-EM structure of the polyphosphate polymerase VTC: Coupling polymer synthesis to membrane transit
Authors: Liu Wei , Wang Jiening , Comte-Miserez Véronique , Zhang Mengyu , Yu Xuejing , Chen Qingfeng , Mayer Andreas , Wu Shan , Ye Sheng 
Published: January 2023

ABSTRACTThe eukaryotic polyphosphate (polyP) polymerase VTC complex synthesizes polyP from adenosine triphosphate (ATP) and translocates polyP across the vacuolar membrane to maintain an intracellular phosphate (Pi) homeostasis. To discover how VTC complex solves this fundamental aspect, we determined a cryo-electron microscopy structure of an endogenous VTC complex (Vtc4/Vtc3/Vtc1) fromSaccharomyces cerevisiaeat 3.1 Å resolution. The structure reveals a heteropentameric architecture of one Vtc4, one Vtc3 and three Vtc1 subunits. The transmembrane region forms a polyP selective channel, probably adopting a resting state conformation, in which a latch-like, horizonal helix of Vtc4 limits the entrance. The catalytic Vtc4 central domain locates on top of the pseudo-symmetric polyP channel, creating a strongly electropositive pathway for nascent polyP that can couple synthesis to translocation. The SPX domain of Vtc4 positively regulates polyP synthesis and regulation of VTC complex. The non-catalytic Vtc3 regulates VTC through a phoshorylatable loop. Our findings, along with the functional data, allow us to propose a mechanism of polyP channel gating and VTC complex activation.
Counterintuitive Binding of Phosphorylated DEP Domain from Dishevelled Protein to Negatively Charged Membranes
Authors: Falginella Francesco L. , Drabinová Martina , Bryja Vítezslav , Vácha Robert 
Published: January 2023

ABSTRACTTo accomplish its role of signaling hub in all Wnt signaling pathways, Dishevelled (DVL) protein needs to dynamically relocalize to the inner leaflet of the cellular plasma membrane (PM). Combined experimental and computational evidence showed that the binding of DVL to the PM is mainly driven by the electrostatic attraction between a stretch of positively charged amino acids located on the C-terminal DEP domain of DVL and anionic phospholipid species, with a striking preference for phosphatidic acid (PA). Here, by means of computational simulations and QCM-D experiments, we demonstrate that four recently identified phosphorylation sites on DEP domain, alter the electrostatic potential of the membrane binding interface, but do not prevent the recruitment to anionic membranes. On the contrary, the phosphorylated residues are involved in hydrogen bond and ion-mediated interactions with the lipid headgroup of PA. Our results suggest that the effect of phosphorylation on protein-membrane association could be counterintuitive and sensitive to changes in the local environment including specific lipids, salts, and pH.SIGNIFICANCEPhosphorylation regulates the cellular activity and localization of many peripheral proteins by, among others, decreasing the affinity for negatively charged membranes. Here, we report how phosphorylation affects the membrane interaction of DEP domain from Dishevelled protein, the intracellular signaling hub in Wnt pathways. We found that despite the negative charge induced by phosphorylation, DEP domain was steadily adsorbed to the surface of negatively charged PA-rich membranes, due to a dense network of cation-mediated interactions and hydrogen bonds.
Carotenoid retention during post-harvest storage ofCapsicum annuum: the role of the fruit surface structure
Authors: Holden Alexandra C. , Cohen Hagai , Daniel V. Rickett , Aharoni Asaph , Fraser Paul D. 
Published: January 2023

AbstractDegradation of carotenoids in food crops during post-harvest storage results in major economic and nutritional losses. In this study, a pepper (Capsicum annuum) panel for post-harvest carotenoid retention was studied to elucidate underlying mechanisms associated with this commercial trait of interest. Quantitative determination of carotenoid pigments and concurrent cellular analysis indicated that those pepper fruit with thicker lipid exocarp layers and smooth surfaces, following post-harvest drying and storage, possessed increased carotenoid retention. Total cutin monomer content increased in high carotenoid retention fruits and sub-epidermal cutin deposits were responsible for the difference in exocarp thickness. Cutin biosynthesis and cuticle precursor transport genes were differentially expressed between high and low carotenoid retention genotypes, and this supports the finding that fruit cuticle biosynthesis is associated with carotenoid retention. Carotenoids were located within cells embedded within the sub-epidermal cutin layer, and these carotenoids were protected from degradation due to the lack of permeability of the fruit surface to reactive oxygen species, and their precursors. The identification of a novel role for the pepper fruit surface in protecting against carotenoid degradation serves as an important discovery for the function of the fruit cuticle and provides an exploitable resource to enhance fruit quality.Highlight statementCarotenoid pigments in Chilli pepper confer post-harvest colour and nutritional quality. Analysis of diverse commercial genotypes indicates the involvement of the fruit surface in carotenoid retention
The Autophagy Receptor NBR1 Directs the Clearance of Photodamaged Chloroplasts
Authors: Lee Han Nim , Chacko Jenu Varghese , Solís Ariadna Gonzalez , Chen Kuo-En , Barros Jessica , Signorelli Santiago , Harvey Millar A. , Vierstra Richard D. , Eliceiri Kevin W. , Otegui Marisa S. 
Published: January 2023

AbstractThe ubiquitin-binding NBR1 autophagy receptor plays a prominent role in recognizing ubiquitylated protein aggregates for vacuolar degradation by macroautophagy. Here, we show that upon exposing Arabidopsis plants to intense light, NBR1 associates with photodamaged chloroplasts independently of ATG7, a core component of the canonical autophagy machinery. NBR1 coats both the surface and interior of chloroplasts, which is then followed by direct engulfment of the organelles into the central vacuole via a microautophagy-type process. The relocalization of NBR1 into chloroplasts does not require the chloroplast translocon complexes embedded in the envelope but is instead greatly enhanced by removing the self-oligomerization mPB1 domain of NBR1. The delivery of NBR1-decorated chloroplasts into vacuoles depends on the ubiquitin-binding UBA2 domain of NBR1 but is independent of the ubiquitin E3 ligases SP1 and PUB4, known to direct the ubiquitylation of chloroplast surface proteins. Compared to wild-type plants,nbr1mutants have altered levels of a subset of chloroplast proteins and display abnormal chloroplast density and sizes upon high light exposure. We postulate that, as photodamaged chloroplasts lose envelope integrity, cytosolic ligases reach the chloroplast interior to ubiquitylate thylakoid and stroma proteins which are then recognized by NBR1 for autophagic clearance. This study uncovers a new function of NBR1 in the degradation of damaged chloroplasts by microautophagy.
Engineered Antiviral Sensor Targets Infected Mosquitoes
Authors: Dalla Benetta Elena , López-Denman Adam J. , Li Hsing-Han , Masri Reem A. , Brogan Daniel J. , Bui Michelle , Yang Ting , Li Ming , Dunn Michael , Klein Melissa J. , Jackson Sarah , Catalan Kyle , Blasdell Kim R. , Tng Priscilla , Antoshechkin Igor , Alphey Luke S. , Paradkar Prasad N. , Akbari Omar S. 
Published: January 2023

AbstractEscalating vector disease burdens pose significant global health risks, so innovative tools for targeting mosquitoes are critical. We engineered an antiviral strategy termed REAPER (vRNAExpressionActivatesPoisonousEffectorRibonuclease) that leverages the programmable RNA-targeting capabilities of CRISPR Cas13 and its potent collateral activity. Akin to a stealthy Trojan Horse hiding in stealth awaiting the presence of its enemy, REAPER remains concealed within the mosquito until an infectious blood meal is up taken. Upon target viral RNA infection, REAPER activates, triggering programmed destruction of its target arbovirus such as chikungunya. Consequently, Cas13 mediated RNA targeting significantly reduces viral replication and its promiscuous collateral activity can even kill infected mosquitoes. This innovative REAPER technology adds to an arsenal of effective molecular genetic tools to combat mosquito virus transmission.One-Sentence SummaryEngineered Cas13-based antiviral sensor kills infected mosquitoes upon activation from arbovirus infection.
Haploid androgenetic development in bovines reveals imbalanced WNT signaling and impaired cell fate differentiation
Authors: Aguila Luis M. , Nociti Ricardo P. , Sampaio Rafael V. , Therrien Jacinthe , Meirelles Flavio V. , Felmer Ricardo N. , Smith Lawrence C. 
Published: January 2023

AbstractHaploid embryos have contributed significantly to our understanding of the role of parental genomes in development and can be applied to important biotechnology for human and animal species. However, development to the blastocyst stage is severely hindered in bovine haploid androgenetic embryos (hAE). To further our understanding of such developmental arrest, we performed a comprehensive comparison of the transcriptomic profile of morula-stage embryos, which were validated by qRT-PCR of transcripts associated with differentiation in haploid and biparental embryos. Among numerous disturbances, results showed that pluripotency pathways, especially the wingless-related integration site (WNT) signaling, were particularly unbalanced in hAE. Moreover, transcript levels ofKLF4, NANOG, POU5F1, SOX2, CDX2, CTNNBL1, AXIN2, andGSK3Bwere noticeably altered in hAE, suggesting disturbance of pluripotency and canonical WNT pathway. To evaluate the role of WNT on hAE competence, we exposed early day-5 morula stage embryos to theGSK3Binhibitor CHIR99021. Although no alterations were observed in pluripotency and WNT-related transcripts, exposure to CHIR99021 improved their ability to reach the blastocysts stage, confirming the importance of the WNT pathway in the developmental features of bovine hAE.Summary statementThis study shows the importance of the WNT pathway on bovine haploid androgenetic development by walking through transcriptomics and pluripotency markers associated with cell fate determination during early development.
Dimerization mechanism of an inverted-topology ion channel in membranes
Authors: Ernst Melanie , Orabi Esam A. , Stockbridge Randy B. , Faraldo-Gómez José D. , Robertson Janice L. 
Published: January 2023

AbstractMany ion channels are multi-subunit complexes with a polar permeation pathway at the oligomeric interface, but their mechanisms of assembly into functional, thermodynamically stable units within the membrane are largely unknown. Here we characterize the assembly of the inverted-topology, homodimeric fluoride channel Fluc, leveraging a known mutation, N43S, that weakens Na+binding to the dimer interface, thereby unlocking the complex. While single-channel recordings show Na+is required for activation, single-molecule photobleaching and bulk Förster Resonance Energy Transfer experiments in lipid bilayers demonstrate that N43S Fluc monomers and dimers exist in dynamic equilibrium, even without Na+. Molecular dynamics simulations indicate this equilibrium is dominated by a differential in the lipid-solvation energetics of monomer and dimer, which stems from hydrophobic exposure of the polar ion pathway in the monomer. These results suggest a model wherein membrane-associated forces induce channel assembly while subsequent factors, in this case Na+binding, result in channel activation.TeaserMembrane morphology energetics foster inverted-topology Fluc channels to form dimers, which then become active upon Na+binding.
Performant web-based interactive visualization tool for spatially-resolved transcriptomics experiments
Authors: Sriworarat Chaichontat , Nguyen Annie , Eagles Nicholas J. , Collado-Torres Leonardo , Martinowich Keri , Maynard Kristen R. , Hicks Stephanie C. 
Published: January 2023

AbstractHigh-resolution and multiplexed imaging techniques are giving us an increasingly detailed observation of a biological system. However, sharing, exploring, and customizing the visualization of large multidimensional images can be a challenge. Here, we introduce Samui, a performant and interactive image visualization tool that runs completely in the web browser. Samui is specifically designed for fast image visualization and annotation and enables users to browse through large images and their selected features within seconds of receiving a link. We demonstrate the broad utility of Samui with images generated with two platforms: Vizgen MERFISH and 10x Genomics Visium Spatial Gene Expression. Samui along with example datasets is available athttps://samuibrowser.com.
Arrhythmia-associated Calmodulin Variants Interact with KCNQ1 to Confer Aberrant Membrane Trafficking and Function
Authors: Kang Po wei , Woodbury Lucy , Angsutararux Paweorn , Sambare Namit , Shi Jingyi , Marras Martina , Abella Carlota , Bedi Anish , Zinn DeShawn , Cui Jianmin , Silva Jonathan R. 
Published: January 2023

AbstractRationaleMissense variants in calmodulin (CaM) predispose patients to arrhythmias associated with high mortality rates. As CaM regulates several key cardiac ion channels, a mechanistic understanding of CaM variant-associated arrhythmias requires elucidating individual CaM variant effect on distinct channels. One key CaM regulatory target is the KCNQ1 (KV7.1) voltage-gated potassium channel that underlie the IKscurrent. Yet, relatively little is known as to how CaM variants interact with KCNQ1 or affect its function.ObjectiveTo observe how arrhythmia-associated CaM variants affect binding to KCNQ1, channel membrane trafficking, and KCNQ1 function.Methods and ResultsWe combine a live-cell FRET binding assay, fluorescence trafficking assay, and functional electrophysiology to characterize >10 arrhythmia-associated CaM variants effect on KCNQ1. We identify one variant (G114W) that exhibits severely weakened binding to KCNQ1 but find that most other CaM variants interact with similar binding affinity to KCNQ1 when compared to CaM wild-type over physiological Ca2+ranges. We further identify several CaM variants that affect KCNQ1 and IKsmembrane trafficking and/or baseline current activation kinetics, thereby contextualizing KCNQ1 dysfunction in calmodulinopathy. Lastly, we delineate CaM variants with no effect on KCNQ1 function.ConclusionsThis study provides comprehensive functional data that reveal how CaM variants contribute to creating a pro-arrhythmic substrate by causing abnormal KCNQ1 membrane trafficking and current conduction. We find that CaM variant regulation of KCNQ1 is not uniform with effects varying from benign to significant loss of function. This study provides a new approach to collecting details of CaM binding that are key for understanding how CaM variants predispose patients to arrhythmia via the dysregulation of multiple cardiac ion channels.
Staphylococcalsecreted cytotoxins are competition sensing signals forPseudomonas aeruginosa
Authors: Wang Grace Z. , Warren Elizabeth A. , Haas Allison L. , Peña Andrea Sánchez , Kiedrowski Megan R. , Lomenick Brett , Chou Tsui-Fen , Bomberger Jennifer M. , Tirrell David A. , Limoli Dominique H. 
Published: January 2023

ABSTRACTCoinfection with two notorious opportunistic pathogens, the Gram-negativePseudomonas aeruginosaand Gram-positiveStaphylococcus aureus, dominates chronic pulmonary infections. While coinfection is associated with poor patient outcomes, the interspecies interactions responsible for such decline remain unknown. Here, we dissected molecular mechanisms of interspecies sensing betweenP. aeruginosaandS. aureus. We discovered thatP. aeruginosasensesS. aureussecreted peptides and, counterintuitively, moves towards these toxins.P. aeruginosatolerates such a strategy through “competition sensing”, whereby it preempts imminent danger/competition by arming cells with type six secretion (T6S) and iron acquisition systems. Intriguingly, while T6S is predominantly described as weaponry targeting Gram-negative and eukaryotic cells, we find that T6S is essential for fullP. aeruginosacompetition withS. aureus, a previously undescribed role for T6S. Importantly, competition sensing was activated during coinfection of bronchial epithelia, including T6S islands targeting human cells. This study reveals critical insight into both interspecies competition and how antagonism may cause collateral damage to the host environment.
Modelling the adoption of Internet of things (IoT) for sustainable construction in a developing economy
Authors: Dosumu Oluwaseun Sunday , Uwayo Seth Moise 
Published: January 2023

PurposeThe need to improve sustainable construction with the appropriate technology adoption in the fourth and fifth industrial revolutions cannot be overemphasized. This study modelled the adoption of Internet of things (IoT) technologies in the construction industry to improve sustainable construction. Consequently, the study would assist construction stakeholders and policymakers in achieving sustainable construction through concerted efforts toward improving IoT adoption.Design/methodology/approachThe study obtained the IoT technologies and the challenges of IoT adoption in the construction industry through a literature review. Subsequently, the awareness (knowledge of the existence and uses) of IoT technologies, the significance of the challenges and the level of adoption of the IoT technologies were tested on construction stakeholders in Kigali with a structured questionnaire. Consequently, a structural equation model of IoT adoption was developed.FindingsThe structural equation model indicates the relationship between the awareness, challenges and adoption of IoT technologies. It equally describes the effect sizes of the awareness and challenges of IoT on the adoption of IoT technologies.Research limitations/implicationsThis study is beneficial to construction stakeholders as it portrays the exclusive impacts of the awareness and the challenges of IoT technologies on the adoption of IoT. The study also benefits the government and policymakers as it provides a basis for intensifying efforts to create awareness and mitigate the challenges of IoT adoption for sustainable construction. The study also provides a basis for future studies on IoT adoption in the construction industry.Originality/valueThere are several studies on the adoption of IoT. There is however scarcity of studies on how the awareness and the challenges of IoT technologies impact the adoption of IoT in the construction industry. Thus, this study bridges the gap by modelling the adoption of IoT technologies for improved sustainable construction.
Protecting workers' well-being in times of crisis: the first wave of COVID-19 in an Australian healthcare setting in 2020
Authors: Achat Helen M. , Stubbs Joanne M. , Mittal Rakhi , Schindeler Suzanne , Gilroy Nicole 
Published: January 2023

PurposeThe COVID-19 pandemic continues to challenge healthcare systems worldwide. The authors examined the lived experiences and perceptions of healthcare workers (HCWs) in managerial and senior positions to explore the pandemic's effects on well-being and valued organisational responses.Design/methodology/approachUsing purposive sampling, the authors conducted 39 semi-structured interviews with senior staff including health service administrators, heads of department and senior clinicians at a designated COVID-19 facility in New South Wales, Australia. Interviews were conducted from November 2020 to February 2021 to reflect on experiences during the height of the pandemic in 2020 (mid-March to the end of May 2020).FindingsWorkplace experiences affecting HCWs' well-being included being shunned by others, fear of infecting family, fear of the unknown, concerns about personal protective equipment, lack of direction from above and increased workload. Organisational interventions to protect the health and safety of HCWs and their patients included redeployment, improved communication, effective management committees, education and mental health supports.Practical implicationsOrganisations can minimise worker-identified factors threatening their well-being during a health crisis by applying broad-ranging initiatives including inclusive and open communication, promoting flexible work practices, providing up-to-date guidelines and policies and fostering camaraderie between workers.Originality/valueThe voices of senior clinical and managerial staff have been largely unheard during the COVID-19 pandemic. The authors addressed this deficit by describing their experiences and insights regarding the pandemic's impact on well-being and the organisation's responses to simultaneously safeguarding its staff and providing quality patient care.
Tribological properties of trimethylolpropane oleate synthesized through esterification reaction with nano-MoS2 catalysts of different shapes
Authors: Li Zhixiang , Han Shuo , Wang Lei , Hu Kunhong 
Published: January 2023

Purpose
This study aims to investigate the catalytic performance and tribological properties of MoS2 powder.

Design/methodology/approach
In this work, the authors attempted to use MoS2 nanoparticles (nano-MoS2) as a catalyst to synthesize trimethylolpropane oleate (TMPTO) by esterification of trimethylolpropane and oleic acid. The small amount of highly dispersed nano-MoS2 catalyst remaining in TMPTO needed not to be separated and could be used as a lubricant modifier directly to achieve the purpose of improving the lubricity performance of TMPTO.

Findings
The results demonstrated that nano-MoS2 had good catalytic esterification ability and achieved in situ dispersion of about 0.191% nano-MoS2 in TMPTO while catalyzing the synthesis of base oil. After high-speed centrifugal sedimentation treatment, the product TMPTO still retained about 0.008% of nano-MoS2. The above-synthesized TMPTO has significantly better lubricity performance than commercially available TMPTO, in which the friction coefficient and wear rate could be reduced by 75%.

Originality/value
The results of this study provide an idea for the design of catalysts for ester oil synthesis.