Flexible Tail Research Articles

CG modeling of nucleosome arrays reveals the salt-dependent chromatin fiber conformational variability.

Eukaryotic DNA is packaged in the cell nucleus into chromatin, composed of arrays of DNA-histone protein octamer complexes, the nucleosomes. Over the past decade, it has become clear that chromatin structure in vivo is not a hierarchy of well-organized folded nucleosome fibers but displays considerable conformational variability and heterogeneity. In vitro and in vivo studies, as well as computational modeling, have revealed that attractive nucleosome-nucleosome interaction with an essential role of nucleosome stacking defines chromatin compaction. The internal structure of compacted nucleosome arrays is regulated by the flexible and dynamic histone N-terminal tails. Since DNA is a highly negatively charged polyelectrolyte, electrostatic forces make a decisive contribution to chromatin formation and require the histones, particularly histone tails, to carry a significant positive charge. This also results in an essential role of mobile cations of the cytoplasm (K+, Na+, Mg2+) in regulating electrostatic interactions. Building on a previously successfully established bottom-up coarse-grained (CG) nucleosome model, we have developed a CG nucleosome array (chromatin fiber) model with the explicit presence of mobile ions and studied its conformational variability as a function of Na+ and Mg2+ ion concentration. With progressively elevated ion concentrations, we identified four main conformational states of nucleosome arrays characterized as extended, flexible, nucleosome-clutched, and globular fibers.

About bacteriophage tail terminator and tail completion proteins: structure of the proximal extremity of siphophage T5 tail.

Bacteriophages are viruses infecting bacteria. The vast majority of them bear a tail, allowing host recognition, cell wall perforation, and DNA injection into the host cytoplasm. Using electron cryo-microscopy (cryo-EM) and single particle analysis, we determined the organization of the tail proximal extremity of siphophage T5 that possesses a long flexible tail and solved the structure of its tail terminator protein p142 (TrP142). It allowed us to confirm the common evolutionary origin between T5 TrPp142 and other known or putative TrPs from siphophages, myophages, and bacterial tail-like machines, despite very poor sequence conservation. By also determining the structure of the T5 tail proximal extremity after interaction with T5 bacterial receptor FhuA, we showed that no conformational changes occur in TrPp142 and confirmed that the infection signal transduction is not carried by the tube itself. We also investigated the location of T5 Neck1 or tail completion protein p143 (TCPp143) and showed, thanks to a combination of cryo-EM and structure prediction using Alphafold2, that it is not located at the capsid-to-tail interface as suggested by its position in the genome, but instead, very unexpectedly, on the side of T5 tail tip, and that it appears to be monomeric. Based on structure comparison with other putative TCPs predicted structures, this feature could not be shared by other TCPs and questions the affiliation of p143 to this family of protein.IMPORTANCEBacteriophages, viruses infecting bacteria, are the most abundant living entities on Earth. They are present in all ecosystems where bacteria develop and are instrumental in the regulation, diversity, evolution, and pathogeny of microbial populations. Moreover, with the increasing number of pathogenic strains resistant to antibiotics, virulent phages are considered a serious alternative or complement to classical treatments. 96% of all phages present a tail that allows host recognition and safe channeling of the DNA to the host cytoplasm. We present the atomic model of the proximal extremity of the siphophage T5 tail, confirming structural similarities with other phages. This structure, combined with results previously published and further explored, also allowed a review and a discussion on the role and localization of a mysterious tail protein, the tail completion protein, which is known to be present in the phage tails, but that was never identified in a phage structure.

Transition metal transporting P-type ATPases: terminal metal-binding domains serve as sensors for autoinhibitory tails.

Copper is an essential micronutrient and yet is highly toxic to cells at elevated concentrations. P1B-ATPase proteins are critical for this regulation, providing active extrusion across cellular membranes. One unique molecular adaptation of P1B-ATPases compared to other P-type ATPases is the presence of metal-binding domains (MBDs) at the cytosolic termini, which however are poorly characterized with an elusive mechanistic role. Here we present the MBD architecture in metal-free and metal-bound forms of the archetype Cu+-specific P1B-ATPase LpCopA, determined using NMR. The MBD is composed of a flexible tail and a structured core with a metal ion binding site defined by three sulfur atoms, one of which is pertinent to the so-called CXXC motif. Furthermore, we demonstrate that the MBD rather than being involved in ion delivery likely serves a regulatory role, which is dependent on the classical P-type ATPase E1-E2 transport mechanism. Specifically, the flexible tail appears responsible for autoinhibition while the metal-binding core is used for copper sensing. This model is validated by a conformation-sensitive and MBD-targeting nanobody that can structurally and functionally replace the flexible tail. We propose that autoinhibition of Cu+-ATPases occurs at low copper conditions via MBD-mediated interference with the soluble domains of the ATPase core and that metal transport is enabled when copper levels rise, through metal-induced dissociation of the MBD. This allows P1B-ATPase 'vacuum cleaners' to tune their own activity, balancing the levels of critical micronutrients in the cells.

Sequential Kalman tuning of the t-preconditioned Crank-Nicolson algorithm: efficient, adaptive and gradient-free inference for Bayesian inverse problems

Abstract Ensemble Kalman Inversion (EKI) has been proposed as an efficient method for the approximate solution of Bayesian inverse problems with expensive forward models. However, when applied to the Bayesian inverse problem EKI is only exact in the regime of Gaussian target measures and linear forward models. In this work we propose embedding EKI and Flow Annealed Kalman Inversion, its normalizing flow (NF) preconditioned variant, within a Bayesian annealing scheme as part of an adaptive implementation of the t-preconditioned Crank-Nicolson (tpCN) sampler. The tpCN sampler differs from standard pCN in that its proposal is reversible with respect to the multivariate t-distribution. The more flexible tail behaviour allows for better adaptation to sampling from non-Gaussian targets. Within our Sequential Kalman Tuning (SKT) adaptation scheme, EKI is used to initialize and precondition the tpCN sampler for each annealed target. The subsequent tpCN iterations ensure particles are correctly distributed according to each annealed target, avoiding the accumulation of errors that would otherwise impact EKI. We demonstrate the performance of SKT for tpCN on three challenging numerical benchmarks, showing significant improvements in the rate of convergence compared to adaptation within standard SMC with importance weighted resampling at each temperature level, and compared to similar adaptive implementations of standard pCN. The SKT scheme applied to tpCN offers an efficient, practical solution for solving the Bayesian inverse problem when gradients of the forward model are not available. Code implementing the SKT schemes for tpCN is available at https://github.com/RichardGrumitt/KalmanMC.

The disordered negatively charged C-terminus of the large HECT E3 ubiquitin ligase HERC2 provides structural and thermal stability to the HECT C-lobe.

Homologous to the C-terminus of E6AP (HECT) and RCC1-like domain (RLD)-containing protein 2 (HERC2) is a large, 528 kDa E3 ubiquitin ligase that is associated with cancer, oculocutaneous albanism type 2, Prader-Willi syndrome, and other neurological diseases. HERC2 has been found to contribute to double-stranded DNA break repairs, tumor suppression, maintaining centrosome architecture, and ubiquitylation. The C-terminal portion of the HECT domain (C-lobe) of HERC2 is responsible for transferring ubiquitin to a substrate but the precise function of the other eight domains in HERC2 are unknown. Interestingly, HERC2 contains a unique and negatively charged C-terminal tail adjoined to the C-lobe that is predicted to act as a linker to promote interactions between HERC2 and its binding partners. This study aims to better understand the function and relevance of HERC2 in disease by investigating the structural aspects of the HERC2 C-lobe and HERC2 C-terminal tail using AlphaFold followed by molecular dynamics (MD) simulations, multidimensional nuclear magnetic resonance (NMR), and circular dichroism (CD). Secondary structure content analysis from MD simulations and the fully resonance assigned 1H-15N HSQC spectra of the HERC2 C-lobe and the isolated C-terminal tail confirm that the C-lobe is well-folded but the C-terminal tail is disordered. CD melting curves indicate that the flexible C-terminal tail provides improved stability to the C-lobe. Additionally, MD simulations have identified that the interaction between residues D4829 and R4728 is prevalent among the non-bonded contacts between the tail and the C-lobe. Overall, our results demonstrate that the negatively charged C-terminal tail is disordered, provides stability to the C-lobe, and may act as a flexible scaffold for protein-protein interactions.

Soluble αβ-tubulins reversibly sequester TTC5 to regulate tubulin mRNA decay

Microtubules, built from heterodimers of α- and β-tubulins, control cell shape, mediate intracellular transport, and power cell division. The concentration of αβ-tubulins is tightly controlled through a posttranscriptional mechanism involving selective and regulated degradation of tubulin-encoding mRNAs. Degradation is initiated by TTC5, which recognizes tubulin-synthesizing ribosomes and recruits downstream effectors to trigger mRNA deadenylation. Here, we investigate how cells regulate TTC5 activity. Biochemical and structural proteomic approaches reveal that under normal conditions, soluble αβ-tubulins bind to and sequester TTC5, preventing it from engaging nascent tubulins at translating ribosomes. We identify the flexible C-terminal tail of TTC5 as a molecular switch, toggling between soluble αβ-tubulin-bound and nascent tubulin-bound states. Loss of sequestration by soluble αβ-tubulins constitutively activates TTC5, leading to diminished tubulin mRNA levels and compromised microtubule-dependent chromosome segregation during cell division. Our findings provide a paradigm for how cells regulate the activity of a specificity factor to adapt posttranscriptional regulation of gene expression to cellular needs.

Combination of a novel bacteriophage and d-serine effectively controls Vibrio parahaemolyticus growth in seafood

Seafood is an important high-protein and low-fat food source prone to spoilage and may harbor food poisoning bacteria. Vibrio parahaemolyticus is a gram-negative halophilic bacterium that causes food poisoning. It is heat labile and can be eliminated through heating; however, V. parahaemolyticus may be present in seafood for raw consumption, such as sashimi. Recently, the use of bacteriophages or phages has been proposed as a non-thermal inactivation method for microbial control. Here, we characterized VF16, a novel phage that infects V. parahaemolyticus, and evaluated the effectiveness of the combination of VF16 with the antimicrobial amino acid d‑serine in controlling V. parahaemolyticus through antimicrobial testing in broth cultures and raw scallops. The morphology of VF16 was examined using transmission electron microscopy, which demonstrated that it had a long, flexible tail characteristic of a siphovirus. Genome sequencing revealed that VF16 is a novel phage belonging to the family Demerecviridae, subfamily Ermolyevavirinae, and genus Vipunavirus. Analysis of adsorption and one-step growth curve in a liquid medium indicated that Ca2+ and Mg2+ are necessary for one or more subsequent replication processes of VF16 but are not essential for its adsorption. The combination of VF16 and d‑serine completely inhibited the regrowth of V. parahaemolyticus in the liquid medium and effectively suppressed its growth in the scallop samples. No VF16-resistant mutants were isolated from the scallop samples treated with VF16 alone or in combination with d‑serine. The combination of phages and d-amino acids is an effective non-thermal inactivation method for controlling foodborne pathogens.

Hopping potential wells and gait switching in a fish-like robot with a bistable tail

Fish outperform current underwater robots in speed, agility, and efficiency of locomotion, in part due to their flexible appendages that are capable of rich combinations of modes of motion. In fish-like robots, actuating many different modes of oscillation of tails or fins can become a challenge. This paper presents a highly underactuated (with a single actuator) fish-like robot with a bistable tail that features a double-well elastic potential. Oscillations of such a tail depend on the frequency and amplitude of excitation, and tuning the frequency–amplitude can produce controllable oscillations in different modes leading to different gaits of the robot. This robot design is inspired by recent work on underactuated flexible swimming robots driven by a single rotor. The oscillations of the rotor can propel and steer the robot, but saturation of the rotor makes performing long turns challenging. This paper demonstrates that by adding geometric bistability to the flexible tail, turns can be performed by controllably exciting single-well oscillations in the tail, while exciting double-well oscillations of the tail produces average straight-line motion. The findings of this paper go beyond the application to a narrow class of fish-like robots. More broadly we have demonstrated the use of periodic excitation to produce bistable response that generate different gaits including a steering gait. The mechanics demonstrated here show the feasibility of applications to other mobile soft robots.

Shape memory liquid crystalline polymers: Stimuli‐responsiveness, advanced technologies, and key applications

AbstractLiquid crystalline polymers (LCPs) represent a distinct class of materials that have garnered significant interest for their utilisation in diverse industrial and engineering applications. A prominent attribute of LCPs is their stimuli‐responsiveness. These materials can undergo deformation and subsequently recover their original shapes when subjected to external stimuli such as heat, light, and electromagnetic fields. The molecular structure of LCPs consists of mesogens and flexible tails, mirroring the fundamental molecular mechanism found in shape memory polymers. This characteristic positions LCPs as promising materials for shape memory applications. This article provides a comprehensive review of LCPs, focusing on their various forms of stimuli‐responsiveness. In addition, it delves into the application of additive manufacturing and machine learning technologies in the context of shape memory LCPs. Finally, the article concludes by exploring the critical applications of LCPs as shape memory materials.

Assembly and activation of EBV latent membrane protein 1

Latent membrane protein 1 (LMP1) is the primary oncoprotein of Epstein-Barr virus (EBV) and plays versatile roles in the EBV life cycle and pathogenesis. Despite decades of extensive research, the molecular basis for LMP1 folding, assembly, and activation remains unclear. Here, we report cryo-electron microscopy structures of LMP1 in two unexpected assemblies: a symmetric homodimer and a higher-order filamentous oligomer. LMP1 adopts a non-canonical and unpredicted fold that supports the formation of a stable homodimer through tight and antiparallel intermolecular packing. LMP1 dimers further assemble side-by-side into higher-order filamentous oligomers, thereby allowing the accumulation and specific organization of the flexible cytoplasmic tails for efficient recruitment of downstream factors. Super-resolution microscopy and cellular functional assays demonstrate that mutations at both dimeric and oligomeric interfaces disrupt LMP1 higher-order assembly and block multiple LMP1-mediated signaling pathways. Our research provides a framework for understanding the mechanism of LMP1 and for developing potential therapies targeting EBV-associated diseases.

Structure-based mapping of the histone-binding pocket of KDM4D using functionalized tetrazole and pyridine core compounds

KDM4 histone demethylases became an exciting target for inhibitor development as the evidence linking them directly to tumorigenesis mounts. In this study, we set out to better understand the binding cavity using an X-ray crystallographic approach to provide a detailed landscape of possible interactions within the under-investigated region of KDM4. Our design strategy was based on utilizing known KDM binding motifs, such as nicotinic acid and tetrazolylhydrazides, as core motifs that we decided to enrich with flexible tails to map the distal histone binding site. The resulting X-ray structures of the novel compounds bound to KDM4D, a representative of the KDM4 family, revealed the interaction pattern with distal residues in the histone-binding site. The most prominent protein rearrangement detected upon ligand binding is the loop movement that blocks the accessibility to the histone binding site. Apart from providing new sites that potential inhibitors can target, the novel compounds may prove helpful in exploring the capacity of ligands to bind in sites distal to the cofactor-binding site of other KDMs or 2-oxoglutarate (2OG)-dependent oxygenases. The case study proves that combining a strong small binding motif with flexible tails to probe the binding pocket will facilitate lead discovery in classical drug-discovery campaigns, given the ease of accessing X-ray quality crystals.

A New Look at the Dirichlet Distribution: Robustness, Clustering, and Both Together

AbstractCompositional data have peculiar characteristics that pose significant challenges to traditional statistical methods and models. Within this framework, we use a convenient mode parametrized Dirichlet distribution across multiple fields of statistics. In particular, we propose finite mixtures of unimodal Dirichlet (UD) distributions for model-based clustering and classification. Then, we introduce the contaminated UD (CUD) distribution, a heavy-tailed generalization of the UD distribution that allows for a more flexible tail behavior in the presence of atypical observations. Thirdly, we propose finite mixtures of CUD distributions to jointly account for the presence of clusters and atypical points in the data. Parameter estimation is carried out by directly maximizing the maximum likelihood or by using an expectation-maximization (EM) algorithm. Two analyses are conducted on simulated data to illustrate the effects of atypical observations on parameter estimation and data classification, and how our proposals address both aspects. Furthermore, two real datasets are investigated and the results obtained via our models are discussed.

Bayesian modal regression based on mixture distributions

Compared to mean regression and quantile regression, the literature on modal regression is very sparse. A unifying framework for Bayesian modal regression is proposed, based on a family of unimodal distributions indexed by the mode, along with other parameters that allow for flexible shapes and tail behaviors. Sufficient conditions for posterior propriety under an improper prior on the mode parameter are derived. Following prior elicitation, regression analysis of simulated data and datasets from several real-life applications are conducted. Besides drawing inference for covariate effects that are easy to interpret, prediction and model selection under the proposed Bayesian modal regression framework are also considered. Evidence from these analyses suggest that the proposed inference procedures are very robust to outliers, enabling one to discover interesting covariate effects missed by mean or median regression, and to construct much tighter prediction intervals than those from mean or median regression. Computer programs for implementing the proposed Bayesian modal regression are available at https://github.com/rh8liuqy/Bayesian_modal_regression.

Alkylamine-tethered molecules recruit FBXO22 for targeted protein degradation

Targeted protein degradation (TPD) relies on small molecules to recruit proteins to E3 ligases to induce their ubiquitylation and degradation by the proteasome. Only a few of the approximately 600 human E3 ligases are currently amenable to this strategy. This limits the actionable target space and clinical opportunities and thus establishes the necessity to expand to additional ligases. Here we identify and characterize SP3N, a specific degrader of the prolyl isomerase FKBP12. SP3N features a minimal design, where a known FKBP12 ligand is appended with a flexible alkylamine tail that conveys degradation properties. We found that SP3N is a precursor and that the alkylamine is metabolized to an active aldehyde species that recruits the SCFFBXO22 ligase for FKBP12 degradation. Target engagement occurs via covalent adduction of Cys326 in the FBXO22 C-terminal domain, which is critical for ternary complex formation, ubiquitylation and degradation. This mechanism is conserved for two recently reported alkylamine-based degraders of NSD2 and XIAP, thus establishing alkylamine tethering and covalent hijacking of FBXO22 as a generalizable TPD strategy.

One-step formation of polymorphous sperm-like microswimmers by vortex turbulence-assisted microfluidics

Microswimmers are considered promising candidates for active cargo delivery to benefit a wide spectrum of biomedical applications. Yet, big challenges still remain in designing the microswimmers with effective propelling, desirable loading and adaptive releasing abilities all in one. Inspired by the morphology and biofunction of spermatozoa, we report a one-step formation strategy of polymorphous sperm-like magnetic microswimmers (PSMs) by developing a vortex turbulence-assisted microfluidics (VTAM) platform. The fabricated PSM is biodegradable with a core-shell head and flexible tail, and their morphology can be adjusted by vortex flow rotation speed and calcium chloride solution concentration. Benefiting from the sperm-like design, our PSM exhibits both effective motion ability under remote mag/netic actuation and protective encapsulation ability for material loading. Further, it can also realize the stable sustain release after alginate-chitosan-alginate (ACA) layer coating modification. This research proposes and verifies a new strategy for the sperm-like microswimmer construction, offering an alternative solution for the target delivery of diverse drugs and biologics for future biomedical treatment. Moreover, the proposed VTAM could also be a general method for other sophisticated polymorphous structures fabrication that isn’t achievable by conventional laminar flow.

Skew Multiple Scaled Mixtures of Normal Distributions with Flexible Tail Behavior and Their Application to Clustering

Skew Multiple Scaled Mixtures of Normal Distributions with Flexible Tail Behavior and Their Application to Clustering

Fully Characterized Effective Bacteriophages Specific against Antibiotic-Resistant Enterococcus faecalis, the Causative Agent of Dental Abscess.

Background and Objectives: Enterococcus faecalis (E. faecalis) is a primary pathogen responsible for dental abscesses, which cause inflammation and pain when trapped between the crown and soft tissues of an erupted tooth. Therefore, this study aims to use specific phages as an alternative method instead of classical treatments based on antibiotics to destroy multidrug-resistant E. faecalis bacteria for treating dental issues. Materials and Methods: In the current study, twenty-five bacterial isolates were obtained from infected dental specimens; only five had the ability to grow on bile esculin agar, and among these five, only two were described to be extensive multidrug-resistant isolates. Results: Two bacterial isolates, Enterococcus faecalis A.R.A.01 [ON797462.1] and Enterococcus faecalis A.R.A.02, were identified biochemically and through 16S rDNA, which were used as hosts for isolating specific phages. Two isolated phages were characterized through TEM imaging, which indicated that E. faecalis_phage-01 had a long and flexible tail, belonging to the family Siphoviridae, while E. faecalis_phage-02 had a contractile tail, belonging to the family Myoviridae. Genetically, two phages were identified through the PCR amplification and sequencing of the RNA ligase of Enterococcus phage vB_EfaS_HEf13, through which our phages shared 97.2% similarity with Enterococcus phage vB-EfaS-HEf13 based on BLAST analysis. Furthermore, through in silico analysis and annotations of the two phages' genomes, it was determined that a total of 69 open reading frames (ORFs) were found to be involved in various functions related to integration excision, replication recombination, repair, stability, and defense. In phage optimization, the two isolated phages exhibited a high specific host range with Enterococcus faecalis among six different bacterial hosts, where E. faecalis_phage-01 had a latent period of 30 min with 115.76 PFU/mL, while E. faecalis_phage-02 had a latent period of 25 min with 80.6 PFU/mL. They were also characterized with stability at wide ranges of pH (4-11) and temperature (10-60 °C), with a low cytotoxic effect on the oral epithelial cell line at different concentrations (1000-31.25 PFU/mL). Conclusions: The findings highlight the promise of phage therapy in dental medicine, offering a novel approach to combating antibiotic resistance and enhancing patient outcomes. Further research and clinical trials will be essential to fully understand the therapeutic potential and safety profile of these bacteriophages in human populations.

Optimal gaits for inertia-dominated swimmers with passive elastic joints.

Animals and some robots locomote by interacting with the environment through cyclic shape changes, or gaits. Many animals make significant use of passive dynamics with flexible tails or pendulum action to reduce the effort required to execute these gaits. Although geometric tools have been developed to study optimal passive gaits for swimmers in drag-dominated physics regimes, they have not yet been used to study larger-scale swimmers whose physics are dominated by inertial effects. In this paper, we leverage previous work in the geometric mechanics field to examine passive-elastic inertial swimmers and show that geometric mechanics can be used to rapidly determine many classes of optimal gaits for such systems. We also discuss how considering swimmer metabolic costs in addition to the mechanical costs of driving actuation is useful for discussing swimmer efficiency. In particular, we focus on two models of active-passive swimming inertial systems: the perfect-fluid three-link swimmer, and a swimmer with a passively flexible tail.

The influence of the side-arms structure on the properties of the liquid crystal dimers centred on mandelic acid

ABSTRACT With mandelic acid (MA) as the chiral core, two series of non-symmetric liquid crystal (LC) dimers have been synthesised by changing the rigidity of the side-arms, namely the DBM-nC2B series and the DBBM-nC2B series. In the DBM-nC2B series, one side-arm is the same, while the length of the flexible tail of the other side-arm increases from 0 to 4. The same is true of the DBBM-nC2B series. Chemical structures and LC properties of the two series LC dimers were characterised by FT-IR, 1H NMR, 13C NMR, DSC, TGA and POM. The results show that the DBM-nC2B series LC dimers display the nematic phase (N), and the DBBM-nC2B series LC dimers exhibit the chiral nematic phase (N*). It shows that the side-arm should have sufficient rigidity for MA to induce the N* phase. In the same series, when the length of the flexible tails of the side-arm increases from 0 to 4, it does not change the mesophase, while it has some effect on the melting temperature (Tm), the clearing temperature (TN(*)I), and mesophase range of the dimers. The molecular length-width ratio (l/w) and dipole moment (μ) were used to explain the variation of Tm and (TN(*)I) of LC dimer.

Phosphorylation of the alpha-I motif in SYMRK drives root nodule organogenesis

Symbiosis receptor-like kinase SYMRK is required for root nodule symbiosis between legume plants and nitrogen-fixing bacteria. To understand symbiotic signaling from SYMRK, we determined the crystal structure to 1.95 Å and mapped the phosphorylation sites onto the intracellular domain. We identified four serine residues in a conserved "alpha-I" motif, located on the border between the kinase core domain and the flexible C-terminal tail, that, when phosphorylated, drives organogenesis. Substituting the four serines with alanines abolished symbiotic signaling, while substituting them with phosphorylation-mimicking aspartates induced the formation of spontaneous nodules in the absence of bacteria. These findings show that the signaling pathway controlling root nodule organogenesis is mediated by SYMRK phosphorylation, which may help when engineering this trait into non-legume plants.