bluuebunny/crossref_metadata_2025_split huggingface dataset:
Co-translational biogenesis of lipid droplet integral membrane proteins
Authors: Leznicki Pawel , Schneider Hayden O. , Harvey Jada V. , Shi Wei Q. , High Stephen 
Published: August 2021

ABSTRACTMembrane proteins destined for lipid droplets (LDs), a major intracellular storage site for neutral lipids, are inserted into the endoplasmic reticulum (ER) and then trafficked to LDs where they reside in a hairpin loop conformation. Here, we show that LD membrane proteins can be delivered to the ER either co- or post-translationally and that their membrane-embedded region specifies pathway selection. The co-translational route for LD membrane protein biogenesis is insensitive to a small molecule inhibitor of the Sec61 translocon, Ipomoeassin F, and instead relies on the ER membrane protein complex (EMC) for membrane insertion. Strikingly, this route can also result in a transient exposure of the short N-termini of LD membrane proteins to the ER lumen, followed by topological rearrangements that enable their transmembrane segment to form a hairpin loop and N-termini to face the cytosol. Our study reveals an unexpected complexity to LD membrane protein biogenesis and identifies a role for the EMC during their co-translational insertion into the ER.SUMMARY STATEMENTInsertion of many lipid droplet membrane proteins into the endoplasmic reticulum (ER) is co-translational, mediated by the ER membrane protein complex (EMC) and involves topology reorientation.
Genetic load and extinction in peripheral populations: the roles of migration, drift and demographic stochasticity
Authors: Sachdeva Himani , Olusanya Oluwafunmilola , Barton Nick 
Published: August 2021

AbstractWe analyse how migration from a large mainland influences genetic load and population numbers on an island, in a scenario where fitness-affecting variants are unconditionally deleterious, and where numbers decline with increasing load. Our analysis shows that migration can have qualitatively different effects, depending on the total mutation target and fitness effects of deleterious variants. In particular, we find that populations exhibit a genetic Allee effect across a wide range of parameter combinations, when variants are partially recessive, cycling between low-load (large-population) and high-load (sink) states. Increased migration reduces load in the sink state (by increasing heterozygosity) but further inflates load in the large-population state (by hindering purging). We identify various critical parameter thresholds at which one or other stable state collapses, and discuss how these thresholds are influenced by the genetic vs. demographic effects of migration. Our analysis is based on a ‘semi-deterministic’ analysis, which accounts for genetic drift but neglects demographic stochasticity. We also compare against simulations which account for both demographic stochasticity and drift. Our results clarify the importance of gene flow as a key determinant of extinction risk in peripheral populations, even in the absence of ecological gradients.
Pancreatic Brsk2 amplifies parasympathetic signals to promote type 2 diabetes
Authors: Xu Rufeng , Wang Kaiyuan , Yao Zhengjian , Jin Li , Pang Jing , Zhou Yuncai , Wang Kai , Liu Dechen , Zhang Yaqin , Sun Peng , Wang Fuqiang , Chang Xiaoai , Li Yating , Wang Shusen , Zhang Yalin , Lin Shuyong , Hu Cheng , Zhu Yunxia , Han Xiao 
Published: August 2021

ABSTRACTThe parasympathetic nervous system (PNS) modulates postprandial glucose metabolism via innervating pancreas; however, its significance in the pathogenesis of type 2 diabetes (T2DM) remains unclear. Here we show that brain-specific serine/threonine-protein kinase 2 (Brsk2), accumulated in obese mouse islets, responds to PNS activation and initiates pre-absorptive insulin release. In inducible mouse models, excessive Brsk2 amplifies parasympathetic signaling to β cells and increases their secretion, ensuing insulin resistance and T2DM. Conversely, Brsk2 inhibition prevents and treats HFD-induced metabolic abnormities via avoiding β-cell oversecretion. Mechanistically, parasympathetic acetylcholine activates cholinergic receptor M3 (Chrm3), then Chrm3 recruits and stabilizes Brsk2, which in turn phosphorylates phospholipase A2 activating protein (Plaa). A Chrm3-Brsk2-Plaa axis stimulates β-cell hypersecretion during both pre-absorptive and absorptive stages in HFD-feeding mice, thus imposing insulin resistance and β-cell dysfunction. Blocking parasympathetic signaling to β cells by Brsk2 protein restoration, autonomic mediation drugs, or vagotomy restricted diabetes development. Moreover, three human BRSK2 variants are associated with hyperinsulinemia, insulin resistance, and T2DM in the Chinese population. These findings reveal that Brsk2 links parasympathetic nervous system to nutrition-overload induced T2DM.
A Network-centric Framework for the Evaluation of Mutual Exclusivity Tests on Cancer Drivers
Authors: Ahmed Rafsan , Erten Cesim , Houdjedj Aissa , Kazan Hilal , Yalcin Cansu 
Published: August 2021

ABSTRACTOne of the key concepts employed in cancer driver gene identification is that of mutual exclusivity (ME); a driver mutation is less likely to occur in case of an earlier mutation that has common functionality in the same molecular pathway. Several ME tests have been proposed recently, however the current protocols to evaluate ME tests have two main limitations. Firstly the evaluations are mostly with respect to simulated data and secondly the evaluation metrics lack a network-centric view. The latter is especially crucial as the notion of common functionality can be achieved through searching for interaction patterns in relevant networks. We propose a network-centric framework to evaluate the pairwise significances found by statistical ME tests. It has three main components. The first component consists of metrics employed in the network-centric ME evaluations. Such metrics are designed so that network knowledge and the reference set of known cancer genes are incorporated in ME evaluations under a careful definition of proper control groups. The other two components are designed as further mechanisms to avoid confounders inherent in ME detection on top of the network-centric view. To this end, our second objective is to dissect the side effects caused by mutation load artifacts where mutations driving tumor subtypes with low mutation load might be incorrectly diagnosed as mutually exclusive. Finally, as part of the third main component, the confounding issue stemming from the use of nonspecific interaction networks generated as combinations of interactions from different tissues is resolved through the creation and use of tissue-specific networks in the proposed framework. The data, the source code and useful scripts are available at: https://github.com/abu-compbio/NetCentric.
Spatial organisation and homeostasis of epithelial lineages at the gastroesophageal junction is regulated by the divergent Wnt mucosal microenvironment
Authors: Kumar Naveen , Gurumurthy Rajendra Kumar , Prakash Pon Ganish , Kurian Shilpa Mary , Wentland Christian , Brinkmann Volker , Mollenkopf Hans-Joachim , Krammer Tobias , Toussaint Christophe , Saliba Antoine-Emmanuel , Biebl Matthias , Juergensen Christian , Wiedenmann Bertram , Meyer Thomas F , Chumduri Cindrilla 
Published: August 2021

AbstractThe gastroesophageal junction (GEJ), where squamous and columnar epithelia meet, is a hotspot for Barrett’s metaplasia development, dysbiosis and carcinogenesis. However, the mechanisms regulating GEJ homeostasis remain unclear. Here, by employing organoids, bulk and single-cell transcriptomics, single-molecule RNA in situ hybridisations and lineage tracing, we identified the spatial organisation of the epithelial, stromal compartment and the regulators that maintain the normal GEJ homeostasis. During development, common KRT8 progenitors generate committed unilineage p63/KRT5-squamous and KRT8-columnar stem cells responsible for the regeneration of postnatal esophagus and gastric epithelium that meet at GEJ. A unique spatial distribution of Wnt regulators in the underlying stromal compartment of these stem cells creates diverging Wnt microenvironments at GEJ and supports their differential regeneration. Further, we show that these tissue-resident stem cells do not possess the plasticity to transdifferentiate to the other lineage with the altered Wnt signals. Our study provides invaluable insights into the fundamental process of GEJ homeostasis and is crucial for understanding disease development.
A computational modeling approach for predicting multicell patterns based on signaling-induced differential adhesion
Authors: Sivakumar Nikita , Warner Helen V. , Peirce Shayn M. , Lazzara Matthew J. 
Published: August 2021

ABSTRACTDifferential adhesion within cell populations enables the emergence of unique patterns in heterogeneous multicellular systems. This process has been previously explored using synthetically engineered heterogenous multicell spheroid systems, in which cell subpopulations engage in bidirectional intercellular signaling to regulate the expression of different cadherins. While engineered cell systems provide excellent experimental tools to observe pattern formation in cell populations, computational models may be leveraged to more systematically explore the key parameters that drive the emergence of different patterns. We developed and validated two- and three-dimensional agent-based models (ABMs) of spheroid patterning for cells engineered with a bidirectional signaling circuit that regulates N- and P-cadherin expression. The model was used to predict how varying initial cell seedings, cadherin induction probabilities, or homotypic adhesion strengths between cells impact spheroid patterning, and unsupervised machine learning techniques were used to map system parameters to unique spheroid patterns. The model was then deployed to design new synthetic cell signaling circuits based on a desired final multicell pattern.
Dynamically unstable ESS in matrix games under time constraints
Authors: Varga Tamás , Garay József 
Published: August 2021

AbstractMatrix games under time constraints are natural extensions of matrix games. They consider the fact that, in addition to the payoff, a pairwise interaction has a further consequence for the contestants. Namely, both players have to wait for some time before becoming fit to participate in a subsequent interaction. Every matrix game can be assigned a continuous dynamical system (the replicator equation) which describes how the frequencies of different phenotypes evolve in the population. One of the fundamental theorems of evolutionary matrix games asserts that the state corresponding to an evolutionarily stable strategy is an asymptotically stable rest point of the replicator equation (Taylor and Yonker 1978, Hofbauer et al. 1979, Zeeman 1980). Garay et al. (2018) and Varga et al. (2020) generalized the statement to two-strategy and, in some particular cases, three- or more strategy matrix games under time constraints. However, the question of whether the implication holds in general remained open. Here examples are provided demonstrating that the answer is no. Moreover, we point out through the rock-scissor-paper game that arbitrary small differences between waiting times can destabilize the rest point corresponding to an ESS. It is also shown that a stable limit cycle can arise around the unstable rest point in a supercritical Hopf bifurcation.Mathematics Subject Classification91A22, 92D15, 92D25, 91A80, 91A05, 91A10, 91A40, 92D40
Mechanism of Tripeptide Trimming by γ-Secretase
Authors: Bhattarai Apurba , Devkota Sujan , Do Hung Nguyen , Wang Jinan , Bhattarai Sanjay , Wolfe Michael S. , Miao Yinglong 
Published: August 2021

AbstractThe membrane-embedded γ-secretase complex processively cleaves within the transmembrane domain of amyloid precursor protein (APP) to produce 37-to-43-residue amyloid β-peptides (Aβ) of Alzheimer’s disease (AD). Despite its importance in pathogenesis, the mechanism of processive proteolysis by γ-secretase remains poorly understood. Here, mass spectrometry and western blotting were used to quantify the efficiency of the first tripeptide trimming step (Aβ49→Aβ46) of wildtype (WT) and familial AD (FAD) mutant APP substrate. In comparison to WT APP, the efficiency of this first trimming step was similar for the I45F, A42T and V46F APP FAD mutants, but substantially diminished for the I45T and T48P mutants. In parallel with biochemical experiments, all-atom simulations using a novel Peptide Gaussian accelerated molecular dynamics (Pep-GaMD) method were applied to investigate tripeptide trimming of Aβ49 by γ-secretase. The starting structure was active γ-secretase bound to Aβ49 and APP intracellular domain (AICD), as generated from our previous study that captured activation of γ-secretase for the initial endoproteolytic cleavage of APP (Bhattarai et al., ACS Cent Sci, 2020, 6:969-983). Pep-GaMD simulations captured remarkable structural rearrangements of both the enzyme and substrate, in which hydrogen-bonded catalytic aspartates and water became poised for tripeptide trimming of Aβ49 to Aβ46. These structural changes required a positively charged N-terminus of endoproteolytic coproduct AICD, which could dissociate during conformational rearrangements of the protease and Aβ49. The simulation findings were highly consistent with biochemical experimental data. Taken together, our complementary biochemical experiments and Pep-GaMD simulations have enabled elucidation of the mechanism of tripeptide trimming by γ-secretase.Significance statementProduction of amyloid β-peptide (Aβ) of Alzheimer’s disease (AD) from its precursor protein requires a series of proteolytic events carried out by the membrane-embedded γ-secretase complex. Mutations in the substrate and enzyme that produce Aβ cause hereditary AD and these mutations affect tripeptide trimming of initially formed long Aβ peptides by γ-secretase. Little is known about the structural mechanism of this trimming process. Here, we have developed a molecular dynamic model for the first step of trimming (Aβ49 to Aβ46) by γ-secretase. Conformational changes in the enzyme-substrate complex to set up this trimming step require the presence of N-terminally charged endoproteolytic cleavage co-product. Computational effects of AD-causing mutations in Aβ49 on the trimming process were validated through biochemical experiments.
Unraveling the Connections between Calreticulin and Myeloproliferative Neoplasms via Calcium Signalling
Authors: Jaiswal Amit , Wang Zhiguo , Zhu Xudong , Ju Zhenyu 
Published: August 2021

AbstractMutations in the form of insertions and deletions (INDEL) in the calreticulin gene lead to essential thrombocythemia which is characterized by the formation of thrombosis. However, the connection between calreticulin INDEL and essential thrombocythemia remains largely elusive. Through combined molecular dynamics simulation and in-vitro studies on the wild type and mutated isoforms of calreticulin, the mechanism underlying the calreticulin INDEL induced essential thrombocythemia was investigated at the molecular level. Our results demonstrate that mutations in exon-9 could lead to significant conformational variations of calreticulin structure and thereby reducing its interaction with calcium ions due to decreased electrostatic contributions. The consequence of mutations on calreticulin’s structural integrity was revealed by identifying the key residues and their roles in calcium binding. Furthermore, mutations implemented by CRISPR-Cas9 in exon-9 showed diminished calcium signaling in HEK-293T cells, which agree well with our in-silico findings. The current study might help in understanding of the interactions between calreticulin exon-9 INDEL and calcium ions mediated by the structural variations of calreticulin. The results provide useful information for designing novel therapeutic approaches targeting essential thrombocythemia.
Mechanism of Tethered Agonist-Mediated Signaling by Polycystin-1
Authors: Pawnikar Shristi , Magenheimer Brenda S. , Munoz Ericka Nevarez , Maser Robin L. , Miao Yinglong 
Published: August 2021

AbstractPolycystin-1 (PC1) is an important unusual G-protein-coupled receptor (GPCR) with 11 transmembrane (TM) domains and its mutations account for 85% of cases of autosomal dominant polycystic kidney disease (ADPKD). PC1 shares multiple characteristics with Adhesion GPCRs. These include a GPCR proteolysis site that autocatalytically divides these proteins into extracellular, N-terminal and membrane-embedded, C-terminal (CTF) fragments, and a tethered agonist (TA) within the N-terminal stalk of the CTF that is suggested to activate signaling. However, the mechanism by which a TA can activate PC1 is not known. Here, we have combined functional cellular signaling experiments of PC1 CTF expression constructs encoding wild type, stalkless and three different ADPKD stalk variants with all-atom Gaussian accelerated molecular dynamics (GaMD) simulations to investigate TA-mediated signaling activation. Correlations of residue motions and free-energy profiles calculated from the GaMD simulations correlated with the differential signaling abilities of wild type and stalk variants of PC1 CTF. They suggested an allosteric mechanism involving residue interactions connecting the stalk, Tetragonal Opening for Polycystins (TOP) domain and putative pore loop in TA-mediated activation of PC1 CTF. Key interacting residues such as N3074-S3585 and R3848-E4078 predicted from the GaMD simulations were validated by new mutagenesis experiments. Together, these complementary analyses have provided novel insights into a TA-mediated activation mechanism of PC1 CTF signaling, which will be important for future rational drug design targeting PC1.Significance StatementMutations of polycystin-1 (PC1) are the major cause (85% of cases) of autosomal dominant polycystic kidney disease (ADPKD), which is the fourth leading cause of kidney failure. PC1 is thought to function as an atypical GPCR, yet the mechanism by which PC1 regulates G-protein signaling remains poorly understood. A significant portion of ADPKD mutations of PC1 encode a protein with defects in maturation or reduced function that may be amenable to functional rescue. In this work, we have combined complementary biochemical and cellular assay experiments and accelerated molecular simulations, which revealed a novel allosteric transduction pathway in activation of the PC1 CTF. Our findings shall facilitate future rational drug design efforts targeting the PC1 signaling function.
Identification of Potent Small Molecule Inhibitors of SARS-CoV-2 Entry
Authors: Mediouni Sonia , Mou Huihui , Otsuka Yuka , Jablonski Joseph Anthony , Adcock Robert Scott , Batra Lalit , Chung Dong-Hoon , Rood Christopher , de Vera Ian Mitchelle S. , Rahaim Ronald , Ullah Sultan , Yu Xuerong , Nguyen Tu-Trinh , Hull Mitchell , Chen Emily , Bannister Thomas D. , Baillargeon Pierre , Scampavia Louis , Farzan Michael , Valente Susana T. , Spicer Timothy P. 
Published: August 2021

ABSTRACTThe severe acute respiratory syndrome coronavirus 2 responsible for COVID-19 remains a persistent threat to mankind, especially for the immunocompromised and elderly for which the vaccine may have limited effectiveness. Entry of SARS-CoV-2 requires a high affinity interaction of the viral spike protein with the cellular receptor angiotensin-converting enzyme 2. Novel mutations on the spike protein correlate with the high transmissibility of new variants of SARS-CoV-2, highlighting the need for small molecule inhibitors of virus entry into target cells. We report the identification of such inhibitors through a robust high-throughput screen testing 15,000 small molecules from unique libraries. Several leads were validated in a suite of mechanistic assays, including whole cell SARS-CoV-2 infectivity assays. The main lead compound, Calpeptin, was further characterized using SARS-CoV-1 and the novel SARS-CoV-2 variant entry assays, SARS-CoV-2 protease assays and molecular docking. This study reveals Calpeptin as a potent and specific inhibitor of SARS-CoV-2 and some variants.
SARS-CoV-2 variants of concern have acquired mutations associated with an increased spike cleavage
Authors: Escalera Alba , Gonzalez-Reiche Ana S. , Aslam Sadaf , Mena Ignacio , Pearl Rebecca L. , Laporte Manon , Fossati Andrea , Rathnasinghe Raveen , Alshammary Hala , van de Guchte Adriana , Bouhaddou Mehdi , Kehrer Thomas , Zuliani-Alvarez Lorena , Meekins David A. , Balaraman Velmurugan , McDowell Chester , Richt Jürgen A. , Bajic Goran , Sordillo Emilia Mia , Krogan Nevan , Simon Viviana , Albrecht Randy A. , van Bakel Harm , Garcia-Sastre Adolfo , Aydillo Teresa 
Published: August 2021

AbstractFor efficient cell entry and membrane fusion, SARS-CoV-2 spike (S) protein needs to be cleaved at two different sites, S1/S2 and S2’ by different cellular proteases such as furin and TMPRSS2. Polymorphisms in the S protein can affect cleavage, viral transmission, and pathogenesis. Here, we investigated the role of arising S polymorphisms in vitro and in vivo to understand the emergence of SARS-CoV-2 variants. First, we showed that the S:655Y is selected after in vivo replication in the mink model. This mutation is present in the Gamma Variant Of Concern (VOC) but it also occurred sporadically in early SARS-CoV-2 human isolates. To better understand the impact of this polymorphism, we analyzed the in vitro properties of a panel of SARS-CoV-2 isolates containing S:655Y in different lineage backgrounds. Results demonstrated that this mutation enhances viral replication and spike protein cleavage. Viral competition experiments using hamsters infected with WA1 and WA1-655Y isolates showed that the variant with 655Y became dominant in both direct infected and direct contact animals. Finally, we investigated the cleavage efficiency and fusogenic properties of the spike protein of selected VOCs containing different mutations in their spike proteins. Results showed that all VOCs have evolved to acquire an increased spike cleavage and fusogenic capacity despite having different sets of mutations in the S protein. Our study demonstrates that the S:655Y is an important adaptative mutation that increases viral cell entry, transmission, and host susceptibility. Moreover, SARS-COV-2 VOCs showed a convergent evolution that promotes the S protein processing.
Assessing spatial segregation of beluga whales (Delphinapterus leucas) in Western Hudson Bay estuaries
Authors: Westdal Kristin , Davies Jeremy , Ferguson Steve 
Published: August 2021

AbstractSegregation of older adult males from females and immature males is known to occur in some beluga whale populations, but it is unclear if adults accompanied by calves segregate in Hudson Bay, where the largest summering population is found. Using imagery from a photographic aerial survey conducted in August 2015, we considered a number of environmental variables that might explain distribution by age class of beluga near two of three main estuaries (Churchill and Seal River) used by Western Hudson Bay belugas in the summer season. Belugas were identified and classified by age manually using an identification decision tree and GPS coordinates were plotted in ArcGIS. Distribution by age class was examined in relation to distance to coastal habitat and bathymetry to test the predation risk hypothesis, sea surface temperature (thermal advantage hypothesis), and extent of river plume (forge-selection hypothesis). Habitat characteristics and the proportion of age classes in both estuaries were similar between age class groups (with and without calves) indicating no segregation and suggesting the environmental data assessed were not driving patterns of distribution and density of age classes at the spatial and temporal scale being investigated. Results provide a greater understanding of spatial patterns of beluga whale habitat use in western Hudson Bay and information useful in conservation and management advice.