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Biology subjects

Ohta, M.

Publications and source records attributed to Ohta, M..

16 recordsLinked to original sources

Resolution-standardized evaluation of ligand atomic coordinates in crystallographic structures using machine learning

Accurate assessment of ligand coordinate-density consistency across different resolutions remains challenging in macromolecular crystallography. We introduce the atomic Box Correlation Coefficient (aBCC), an atom-level metric for evaluating the consistency between ligand atomic coordinates and electron density in a resolution-standardized framework. To predict aBCC values from electron-density maps, we developed QAEmap, a machine-learning model based on three-dimensional convolutional neural networks (3D-CNNs). The model was trained using Fourier-truncated electron-density maps and corresponding ligand coordinates generated from high-resolution structures in the Protein Data Bank. It was evaluated using both Fourier-truncated electron-density maps and experimentally determined PDB structures. was evaluated using both Fourier-truncated electron-density maps and experimentally determined PDB structures.The prediction accuracy gradually decreased with decreasing resolution, but remained reliable up to [~]3.5 [A]. These results demonstrate that aBCC enables resolution-standardized atom-wise evaluation of coordinate-density consistency across different resolutions and provide a foundation for further development and refinement of machine learning-based coordinate validation. SynopsisWe introduce the atomic box correlation coefficient (aBCC), a machine learning-based metric for the resolution-standardized atom-level evaluation of ligand coordinate-density consistency in crystallographic structures. aBCC provides a common framework for assessing and communicating the local coordinate reliability between structural biologists and researchers in structure-based drug discovery.

molecular biology↗

Using herbarium genomics to understand the history of a global plant invasion

Invasive plants are dynamic eco-evolutionary systems characterized by rapid spread and evolutionary change. Herbarium genomics offers a powerful way to study these processes across broad spatial and temporal scales. We generated low-coverage shotgun-sequencing data from 152 herbarium specimens of the invasive Japanese knotweed species complex (Reynoutria), collected across its native range in Japan and China, and its introduced ranges in Europe and North America, spanning 200 years of global spread. Introduced populations of R. japonica, R. japonica var. compacta and R. sachalinensis showed reduced genetic diversity compared to native populations and were genetically most similar to Japanese specimens, indicating Japan as the source of all three introductions. European and North American R. japonica were genetically highly similar and closely related to local R. x bohemica hybrids, suggesting that these hybrids originated post-introduction, through hybridization and subsequent introgression from the R. japonica parent. A few R. x bohemica hybrids in the UK and France shared a multilocus lineage with R. sachalinensis, indicating an independent but ecologically less successful hybridization event involving introgression from R. sachalinensis. A genetically distinct R. x bohemica from Japan was closely related to a Japanese R. sachalinensis specimen, suggesting that hybridization also occurs in the native range. Introduced R. japonica has remained genetically uniform for [~]200 years reflecting long-term founder effects. The dominance of a single R. japonica lineage supports the general-purpose genotype hypothesis and the importance of vegetative reproduction in its spread. Herbarium genomics thus uncovers the origin and global spread of Japanese knotweed, providing direct molecular evidence of long-term plant invasions.

evolutionary biology↗

Phosphorylation remodels the mitotic centrosome matrix to generate bipartite gamma-tubulin complex docking sites

Mitotic centrosomes consist of centrioles surrounded by a proteinaceous matrix that docks and activates {gamma}-tubulin complexes ({gamma}TuCs) to nucleate microtubules for spindle assembly. During mitotic entry, phosphorylation at centrosomes remodels CDK5RAP2 family matrix proteins to generate {gamma}TuC docking sites. We address the mechanism of this conversion using C. elegans SPD-5 as a model. We show that SPD-5 contains two regions, PRGB1 and PRGB2, that are each sufficient for Polo-Like Kinase 1 (PLK1) phosphorylation-regulated {gamma}TuC binding. We define key phosphosites in each region and uncover autoinhibition mediated by interactions within and between them. PRGB2 is dimeric and requires {gamma}TuCs containing the Mozart family microprotein MZT-1 for binding, whereas PRGB1 is monomeric and binds independently of MZT-1. Our results support PLK1 phosphorylation inducing a conformational change that enables MZT-1-dependent PRGB2 binding, which in turn relieves PRGB1 inhibition. Such a multi-step mechanism would ensure robust spindle assembly by restricting microtubule nucleation in space and time.

cell biology↗

intDesc-AbMut: Describing and understanding how antibody mutations impact their environmental interactions

Previously, we proposed a double-point mutation (DPM) strategy involving the simultaneous substitution of two amino acids to optimize antibodies. By selecting mutants based on the criterion that favorable interactions between mutated residues and their local environments are preserved or enhanced, improvements in antibody affinity were achieved. Nonetheless, manual extraction of these interactions proved to be time-consuming and labor-intensive. Thus, to streamline this process, we developed intDesc-AbMut, an automated software tool designed to extract interactions between designated mutant residues and their surroundings from three-dimensional antigen-antibody complex structures. intDesc-AbMut identifies and classifies 36 distinct types of interactions, enabling visualization of changes before and after mutation. Additionally, the effects of mutations can be represented by interaction descriptors generated from the extracted interactions. As a practical application, we constructed a machine learning model using these descriptors combined with side-chain rotamer frequencies to determine whether the environment around mutated residues in antigen-antibody complexes is crystal-like, achieving a Matthews correlation coefficient of 0.505 and an accuracy of 0.855. The intDesc-AbMut software is available at: https://github.com/riken-yokohama-AI-drug/intDesc/tree/intDesc-AbMut.

bioengineering↗

Cancer-Associated USP28 Missense Mutations Disrupt 53BP1 Interaction and p53 Stabilization

Cellular stress response pathways are essential for genome stability and are frequently dysregulated in cancer. Following mitotic stress, the ubiquitin-specific protease 28 (USP28) and the p53-binding protein 1 (53BP1) form a stable, heritable complex to stabilize the tumor suppressor p53, triggering cell cycle arrest or apoptosis. Here we determine the mechanism by which USP28 stabilizes p53 and show that USP28 is required not only for an efficient stress response but also for maintaining basal p53 levels in some cancer cells. Loss of functional USP28 allows cells to evade mitotic stress and DNA damage responses in a manner that is specific to cell type and cancer context. We identify a prevalent, shorter USP28 isoform critical for p53 stabilization. Its C-terminal domain mediates PLK1-dependent binding to 53BP1, a dimerization-driven interaction necessary for mitotic stress memory, p53 stabilization, and cell cycle arrest. Cancer-associated missense mutations in this domain disrupt 53BP1 binding, impair nuclear localization, and destabilize USP28, compromising p53 stabilization. Notably, mutations in the 53BP1-binding domain occur more frequently in tumors than those in the catalytic domain, suggesting a potential role in cancer progression and implications for therapeutic strategies.

cell biology↗

TRIM37 employs peptide motif recognition and substrate-dependent oligomerization to prevent ectopic spindle pole assembly

Tightly controlled duplication of centrosomes, the major microtubule-organizing centers of animal cells, ensures bipolarity of the mitotic spindle and accurate chromosome segregation. The RBCC (RING-B-box-coiled coil) ubiquitin ligase TRIM37, whose loss is associated with elevated chromosome missegregation and the tumor-prone developmental human disorder Mulibrey nanism, prevents the formation of ectopic spindle poles that assemble around structured condensates containing the centrosomal protein centrobin. Here, we show that TRIM37s TRAF domain, unique in the extended TRIM family, engages peptide motifs in centrobin to suppress condensate formation. TRIM proteins form anti-parallel coiled-coil dimers with RING-B-box domains on each end. Oligomerization due to RING-RING interactions and conformational regulation by B-box-2-B-box-2 interfaces are critical for TRIM37 to suppress centrobin condensate formation. These results indicate that, analogous to anti-viral TRIM ligases, TRIM37 activation is linked to the detection of oligomerized substrates. Thus, TRIM37 couples peptide motif recognition and substrate-dependent oligomerization to effect ubiquitination-mediated clearance of ectopic centrosomal protein assemblies.

cell biology↗

A simple inland culture system provides insights into ascidian post-embryonic developmental physiology

Maintenance and breeding of experimental organisms are fundamental to life sciences, but both initial and running costs, and hands-on zootechnical demands can be challenging for many laboratories. Here, we aimed to further develop a simple protocol for reliable inland culture of tunicate model species of the Ciona genus. We cultured both Ciona robusta and Ciona intestinalis in controlled experimental conditions, with a focus on dietary variables, and quantified growth and maturation parameters. From statistical analysis of these standardized datasets, we gained insights into the post-embryonic developmental physiology of Ciona, and inferred an improved diet and culturing conditions for sexual maturation. We showed that body length is a critical determinant of both somatic and sexual maturation, which suggests the existence of systemic control mechanisms of resource allocation toward somatic growth or maturation, and supports applying size selection as a predictor of reproductive fitness in our inland culture, to keep the healthiest animals at low density in the system. In the end, we successfully established a new protocol, including size selection, to promote both sperm and eggs production. Our protocol using small tanks will empower researchers to initiate inland Ciona cultures with low costs and reduced space constraints.

developmental biology↗

SARS-CoV-2 Omicron XBB.1.5 may be a cautionary variant by in silico study

In this research, we aimed to predict the relative risk of the recent new variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on the basis of our previous research. We first performed molecular docking simulation analyses of the spike proteins with human angiotensin-converting enzyme 2 (ACE2) to determine the binding affinities to human cells of three new variants of SARS-CoV-2: Omicron BQ.1, XBB, and XBB.1.5 We then investigated the three variants to discover the evolutionary distance of the spike protein gene (S gene) from the Wuhan, Omicron BA.1, and Omicron BA.4/5 variants, to understand the changes in the S gene. The results indicated that the XBB.1.5 variant had the highest binding affinity of the spike protein with ACE2 and the longest evolutionary distance of the S gene. This in silico evidence suggested that the XBB.1.5 variant may produce infections that spread more widely and faster than can infections of preexisting variants.

bioinformatics↗

PLK-1 tethered on BUB-1 directs CDC-20 kinetochore recruitment to ensure timelyembryonic mitoses

During mitosis chromosomes assemble kinetochores in order to dynamically couple with spindle microtubules (Cheeseman, 2014; Musacchio & Desai, 2017). Kinetochores also function as signaling hubs directing mitotic progression by recruiting and controlling the fate of the Anaphase Promoting Complex/Cyclosome (APC/C) activator CDC-20 (Lara-Gonzalez et al., 2017; Lara-Gonzalez, Pines, et al., 2021; Musacchio, 2015). Kinetochores either incorporate CDC-20 into checkpoint complexes that inhibit the APC/C or dephosphorylate CDC-20, which allows it to interact with and activate the APC/C (Kim et al., 2017; Lara-Gonzalez et al., 2017). The importance of these two CDC-20 fates likely depends on biological context. In somatic cells the major mechanism controlling mitotic progression is the spindle checkpoint. By contrast, progression through mitosis during the cell cycles of early embryos is largely checkpoint-independent (Clute & Masui, 1995; Duro & Nilsson, 2021; Gerhart et al., 1984; Zhang et al., 2015). Here, by manipulating CDC-20 phosphorylation status, we show that CDC-20 phosphoregulation controls mitotic duration in the C. elegans embryo and defines a checkpoint-independent temporal mitotic optimum for robust embryogenesis. Flux of CDC-20 through kinetochores for local dephosphorylation requires an ABBA motif on BUB-1 that directly interfaces with the structured WD40 domain of CDC-20 (Di Fiore et al., 2015; Diaz-Martinez et al., 2015; He et al., 2013; Kim et al., 2017). We show that a conserved "STP" motif in BUB-1 that docks the mitotic kinase PLK-1 (Qi et al., 2006) is also necessary to recruit CDC-20 to kinetochores and for timely mitotic progression. The kinase activity of PLK-1 is required for CDC-20 to localize to kinetochores and targets a site within the CDC-20-binding ABBA motif of BUB-1; phosphorylation of this site promotes BUB-1-CDC-20 interaction and mitotic progression. Thus, the BUB-1-bound pool of PLK-1 ensures timely mitosis during embryonic cell cycles by promoting CDC-20 recruitment to the vicinity of kinetochore-localized phosphatase activity.

cell biology↗

SARS-CoV-2 Omicron BA.2.75 variant may be much more infective than preexisting variants

ObjectivesIn our previous research, we developed a mathematical model via molecular simulation analysis to predict the infectivity of seven SARS-CoV-2 variants. In this report, we aimed to predict the relative risk of the recent new variants of SARS-CoV-2 as based on our previous research. MethodsWe subjected Omicron BA.4/5 and BA.2.75 variants of SARS-CoV-2 to the analysis to determine the absolute evolutionary distance of the spike protein gene (S gene) of the variants from the Wuhan variant so as to appreciate the changes in the spike protein. We performed the molecular docking simulation analyses of the spike proteins with human angiotensin-converting enzyme 2 (ACE2) to understand the docking affinities of these variants. We then compared the evolutionary distances and the docking affinities of these variants with those of the seven variants that we had analyzed in our previous research. ResultsThe evolutionary distances of the S gene in BA.4/5 and BA.2.75 from the Wuhan variant were longer than those of the other variants. BA.2.75 had the highest docking affinity of the spike protein with ACE2 (ratio per Wuhan variant). ConclusionThe important results from this analysis are the following: BA.2.75 has both the highest docking affinity and the longest evolutionary distance of the S gene. These results suggest that BA.2.75 infection can spread farther than can infections of preexisting variants.

bioinformatics↗

Comparing the Effect of Helical-centerline Stent Placement on Blood Flow Velocity with a Straight Stent

Stent treatment can be used to treat blood vessel stenosis in a less invasive manner, but re-stenosis is a concern. Because a helical-stent configuration has been thought to reduce the amount of intimal hyperplasia, the helical stent is considered clinically effective. The effects of the helical stent on blood flow velocity, however, have not been studied. In this study, we estimated flow velocities before and after helical stenting using time-intensity-curve (TIC) from angiography images and compared them with straight stenting velocities. As a result, in all cases (N = 3), the velocity reduction was less with helical stenting than with straight stenting. Based on angiography images, this flow estimation method can estimate patient-specific blood flow velocity in situ even in a presence of a stent.

bioengineering↗

gr Predictor: a Deep-Learning Model for Predicting the Hydration Structures around Proteins

Among the factors affecting biological processes such as protein folding and ligand binding, hydration, which is represented by a three-dimensional water-site-distribution-function around the protein, is crucial. The typical methods for computing the distribution functions, including molecular dynamics simulations and the three-dimensional reference interaction site model (3D-RISM) theory, require a long computation time from hours to tens of hours. Here, we propose a deep-learning model rapidly estimating the distribution functions around proteins obtained by the 3D-RISM theory from the protein 3D structure. The distribution functions predicted using our deep-learning model are in good agreement with those obtained by the 3D-RISM theory. Particularly, the coefficient of determination between the distribution function obtained by the deep-learning model and that obtained using the 3D-RISM theory is approximately 0.98. Furthermore, using a graphics processing unit (GPU), the calculation by the deep learning model is completed in less than one minute, more than 2 orders of magnitude faster than the calculation time of 3D-RISM theory. Therefore, our deep learning model provides a practical and efficient way to calculate the three-dimensional water-site-distribution-functions. The program called "gr Predictor" is available under the GNU General Public License from https://github.com/YoshidomeGroup-Hydration/gr-predictor. Table of Contents graphic O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=55 SRC="FIGDIR/small/488616v1_ufig1.gif" ALT="Figure 1"> View larger version (14K): org.highwire.dtl.DTLVardef@108b3c7org.highwire.dtl.DTLVardef@52ec18org.highwire.dtl.DTLVardef@a388borg.highwire.dtl.DTLVardef@10654ad_HPS_FORMAT_FIGEXP M_FIG C_FIG

biophysics↗

Prediction of infectivity of SARS-CoV2: Mathematical Model with Docking Simulation analysis between Spike Protein and ACE2

Variants of a coronavirus (SARS-CoV-2) have been spreading in a global pandemic. Improved understanding of the infectivity of future new variants is important so that effective countermeasures against them can be quickly undertaken. In our research reported here, we aimed to predict the infectivity of SARS-CoV-2 by using a mathematical model with molecular simulation analysis, and we used phylogenetic analysis to determine the evolutionary distance of the spike protein gene (S gene) of SARS-CoV-2. We subjected the six variants and the wild type of spike protein and human angiotensin-converting enzyme 2 (ACE2) to molecular docking simulation analyses to understand the binding affinity of spike protein and ACE2. We then utilized regression analysis of the correlation coefficient of the mathematical model and the infectivity of SARS-CoV-2 to predict infectivity. The evolutionary distance of the S gene correlated with the infectivity of SARS-CoV-2 variants. The coefficient of the mathematical model obtained with results of molecular docking simulation also correlated with the infectivity of SARS-CoV-2 variants. These results suggest that the data from the docking simulation for the receptor binding domain of variant spike proteins and human ACE2 were valuable for prediction of SARS-CoV-2 infectivity. In addition, we developed a mathematical model for prediction of SARS-CoV-2 variant infectivity by using binding affinity obtained via molecular docking and the evolutionary distance of the S gene.

bioinformatics↗

Intrinsic bursts facilitate learning of Levy flight movements in recurrent neural network models

Isolated spikes and bursts of spikes are thought to provide the two major modes of information coding by neurons. Bursts are known to be crucial for fundamental processes between neuron pairs, such as neuronal communications and synaptic plasticity. Deficits in neuronal bursting can also impair higher cognitive functions and cause mental disorders. Despite these findings on the roles of bursts, whether and how bursts have an advantage over isolated spikes in the network-level computation remains elusive. Here, we demonstrate in a computational model that not isolated spikes but intrinsic bursts can greatly facilitate learning of Levy flight random walk trajectories by synchronizing burst onsets across neural population. Levy flight is a hallmark of optimal search strategies and appears in cognitive behaviors such as saccadic eye movements and memory retrieval. Our results suggest that bursting is a crucial component of sequence learning by recurrent neural networks in the brain.

neuroscience↗

Polo-like kinase 1 independently controls microtubule-nucleating capacity and size of the centrosome

Centrosomes are composed of a centriolar core surrounded by a pericentriolar material (PCM) matrix that docks microtubule-nucleating {gamma}-tubulin complexes. During mitotic entry, the PCM matrix increases in size and nucleating capacity in a process called centrosome maturation. Polo-like kinase 1 (PLK1) localizes to centrosomes and phosphorylates PCM matrix proteins to drive their self-assembly, which leads to PCM expansion; this expansion has been assumed to passively increase microtubule nucleation to support spindle assembly. Here, we show that PLK1 directly controls the generation of binding sites for {gamma}-tubulin complexes on the PCM matrix, independently of PCM expansion. Selective inhibition of PLK1-dependent {gamma}-tubulin docking leads to spindle defects and impaired chromosome segregation, without affecting PCM expansion, highlighting the importance of phospho-regulated centrosomal {gamma}-tubulin docking sites in spindle assembly. Inhibiting both {gamma}-tubulin docking and PCM expansion by mutating substrate target sites fully accounts for the actions of PLK-1 in transforming the centrosome during mitotic entry. Summary StatementPolo-like kinase 1-mediated physical expansion of centrosomes during mitotic entry is proposed to passively increase their microtubule nucleating capacity. Ohta et al. show instead that generation of microtubule-nucleating sites is directly controlled by Polo-like kinase 1, independently of centrosome size.

cell biology↗

The expansion in lymphoid organs of IL-4+ BATF+ T follicular helper cells is linked to IgG4 class switching in vivo

Distinct TFH subsets that influence specific class-switching events are assumed to exist, but the accumulation of isotype-specific TFH subsets in secondary and tertiary lymphoid organs has not been hitherto demonstrated. IL-4 expressing TFH cells are surprisingly sparse in human secondary lymphoid organs. In sharp contrast, in IgG4-related disease (IgG4-RD), a disorder characterized by polarized Ig class switching, most TFH cells in tertiary and secondary lymphoid organs make IL-4. Human IL-4+ TFH cells do not express GATA-3 but express nuclear BATF, and the transcriptomes of IL-4 secreting TFH cells differ both from PD1hi TFH cells that do not secrete IL-4 and IL4-secreting non-TFH cells. Unlike IgG4-RD, IL-4+ TFH cells are rarely found in tertiary lymphoid organs in Sjogrens syndrome, a disorder in which IgG4 is not elevated. The proportion of CD4+IL-4+BATF+ T cells as well as of CD4+IL-4+CXCR5+ T cells in IgG4-RD tissues correlates tightly with tissue IgG4 plasma cell numbers and plasma IgG4 levels in patients but not with the total plasma levels of other isotypes. These data describe a disease-related TFH sub-population in human tertiary and secondary lymphoid organs that is linked to IgG4 class switching.

immunology↗