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Matsushita, T.

Publications and source records attributed to Matsushita, T..

16 recordsLinked to original sources

Evaluating Lightweight and Full Fine-Tuning Strategies Against Classical Machine Learning for Protein Function Prediction

Motivation Protein language models (PLMs) have emerged as powerful tools for sequence-based prediction of protein function, yet systematic benchmarks comparing frozen embeddings, fine-tuning strategies like Low-Rank Adaptation (LoRA) and classical machine learning (ML) remain limited. We benchmarked four ML strategies: ML using amino acid descriptors (SL-AAFeat), ML using frozen embeddings from 20 PLMs across various pooling strategies (SL-Embed), full model fine-tuning (FT-Full) and LoRA-based fine-tuning (FT-LoRA). Performance was evaluated on the in-house VHH phage display dataset (VHH) for binding affinity prediction and the TAPE fluorescence dataset (FLS and FLS10) for mutational effect prediction. Results Model performance depended strongly on the dataset and adaptation strategy. Max pooling consistently improved embedding-based models, while amino acid descriptors remained competitive under specific datasets and resource constraints. Fine-tuning generally provided the highest predictive performance, but the advantage is not universal. Hyperparameter optimization significantly enhanced FT-LoRA, enabling it to outperform FT-Full on the VHH dataset with less than 10% model parameter adaptation. In contrast, FT-Full achieved the best performance on FLS and FLS10. Several medium-sized PLMs performed comparably to larger models, highlighting favorable performance-efficiency trade-offs. Overall, this paper presents a thorough review of PLM utilization strategies and practical recommendations for selecting suitable strategies based on dataset characteristics and available computational resources. Availability The source code used in this manuscript is available in a Zenodo repository at https://doi.org/10.5281/zenodo.21466255.

bioinformatics↗

High-Coverage Jomon Genomes Provide Insights into Population Structure and Genetic Traits of Ancient Japanese Hunter-Gatherers

We analyzed eight high-coverage Jomon genomes spanning 9,000-2,800 years ago across the Japanese archipelago. Population genomic analyses revealed substantial genetic homogeneity across time and space reflecting post-glacial isolation, yet temporal stratification emerged: Initial Jomon (Iyai) individuals formed the earliest branch with weaker affinities to present-day Japanese. Using genomes from Todoroki and Chidorikubo sites, we estimated Jomon ancestry in mainland Japanese at [~]20%, substantially higher than previous estimates ([~]13%). Polygenic score analyses, based on the direct analysis of Jomon genomes, provided the genetic evidence for their phenotypic predispositions, including elevated BMI and metabolic trait scores, reduced immune-related scores, and larger cardiac dimensions, patterns consistent with hunter-gatherer adaptations. These ancestral genetic influences, including elevated risks for obesity and dyslipidemia, persist in present-day Japanese. Our findings demonstrate that Jomon populations maintained genetic continuity despite ecological diversity, while their greater-than-expected contribution to present-day Japanese reshapes population history models and reveals how ancient adaptations continue influencing contemporary disease susceptibility.

genomics↗

A blue light receptor CRY1 regulates de novo shoot regeneration

Tissue culture enables plant transformation and gene editing, yet inefficient regeneration remains a major bottleneck, and the downstream circuitry linking light perception to tissue culture has not been investigated. Here we demonstrate that the blue-light photoreceptor CRYPTOCHROME 1 (CRY1) enhances Arabidopsis thaliana shoot regeneration in a blue light dependent manner. In a two-step root-to-shoot tissue culture system, cry1 mutant exhibits reduced shoot regeneration. Transcriptomic analyses in the cry1 mutant reveal that CRY1 suppresses the expression of AUXIN RESPONSE TRANSCRIPTION FACTOR 3 (ARF3). Consistently, arf3 mutant exhibits enhanced shoot regeneration phenotype. Additionally, genetic and protein interaction assays show that ARF3 acts downstream of CRY1 at both transcriptional and post-translational levels. Furthermore, transcriptomic analyses in the arf3 mutant show that ARF3 activates salicylic acid (SA)-responsive genes that suppress shoot formation, and exogenous SA inhibits regeneration in arf3 calli. Integrating RNA-seq with cis-motif analysis identifies stress-responsive ARF3 targets within the CRY1 network. Our results establish a novel CRY1-ARF3 regulatory module that links blue-light perception to hormonal signaling during regeneration, revealing how plants integrate environmental cues with growth-defense trade-offs. SynopsisThe blue-light photoreceptor CRY1 promotes de novo shoot regeneration in Arabidopsis thaliana by restraining ARF3-mediated SA signaling. It reveals a fundamental trade-off between defense responses and regenerative growth in plants, coordinated by CRY1-ARF3 module. O_LICRY1-mediated blue light signaling enhances de novo shoot regeneration from callus. C_LIO_LIARF3 acts downstream of CRY1 and CRY1 physically interacts with ARF3. C_LIO_LIARF3-activated SA pathway represses shoot regeneration. C_LIO_LICRY1 enhances shoot regeneration by suppressing ARF3-activated stress responsive genes. C_LI

plant biology↗

A nuclear CobW/WW-domain factor represses the CO2-concentrating mechanism in the green alga Chlamydomonas reinhardtii

Microalgae induce a CO2-concentrating mechanism (CCM) to maintain photosynthesis when dissolved CO2 is limited, but how this energy-intensive system is suppressed when CO2 levels rise has remained unclear. The CCM consumes 15-30% of photosynthetically-generated ATP, making its regulation critical for cellular energy balance. Here, we identify a nuclear repressor of the CCM in the green alga Chlamydomonas reinhardtii. A pull-down screen for interacting partners of the master activator CCM1/CIA5 revealed an uncharacterized protein that co-purifies with CCM1 even after high-salt washes. This protein, designated CCM1-binding protein 1 (CBP1), combines a CobW/CobW_C GTP-binding metallochaperone module with a WW domain characteristic of protein-protein interactions. CBP1 colocalizes with CCM1 in the nucleus regardless of CO2 conditions, and the two proteins interact in vivo. CRISPR/Cas9-based disruption of CBP1 does not affect growth or CCM induction under CO2 limitation but derepresses 27 of 41 CCM1-dependent low-CO2 inducible genes under high-CO2 conditions. These include the periplasmic and intracellular carbonic anhydrases (CAH1, LCIB) and inorganic carbon transporters/channels (LCIA, LCI1, BST1, BST3). Consistently, cbp1 mutants accumulate higher levels of CAH1 and LCIB proteins and exhibit a 40% increase in inorganic carbon affinity under high-CO2 conditions; this phenotype is rescued by CBP1 complementation or by acetazolamide treatment. These results demonstrate that CBP1 prevents unnecessary CCM activity when CO2 is abundant, acting upstream of both transporter/channel and carbonic anhydrase modules. Our findings suggest a regulatory mechanism potentially linking zinc-dependent protein chemistry to CCM gene repression, providing insights into energy-efficient CO2 sensing in aquatic photosynthetic organisms. Significance statementAlgae flourish by activating a CO2-concentrating mechanism (CCM) when dissolved CO2 is limited but must deactivate it when CO2 levels increase to conserve energy. We have identified the nuclear protein that functions as this long sought "off switch" in the model green alga. Deletion of this protein causes cells to overproduce CCM transporters and enzymes, maintaining CCM activity even under high CO2 conditions, demonstrating its essential role in suppressing the system when carbon is abundant. This finding illuminates how algae balance energy consumption with carbon capture and offers a new target for engineering strains that fix CO2 more efficiently for biofuel production or climate-mitigation technologies.

plant biology↗

Astringent flavanol fires the locus-noradrenergic system, regulating neurobehavior and autonomic nerves

Astringent flavanols (FLs) have shown enhanced cognitive function in a large intervention trial, but their mechanism remains unclear due to poor bioavailability. An oral dose of FLs increased spontaneous locomotion and memory, with activation of the sympathetic-adrenergic-medullary (SAM) and hypothalamic-pituitary-adrenal (HPA) axis, as shown by increased urinary catecholamine concentrations and corticotropin-releasing hormone (CRH) mRNA expression in the paraventricular nucleus (PVN) of mice. MS imaging showed high intensity of noradrenaline (NA) in the locus coeruleus (LC), which plays a key role in learning and memory, the hypothalamus lateral preoptic area, which affects sleep and arousal, and the brainstem as the origin of the sympathetic nervous system, immediately after FLs administration. Subsequent in situ hybridization (ISH) analysis showed that these NAs originated from LC. Astringent compounds like FLs influence brain function and physiological changes. Furthermore, the present series of results presents novel mechanisms by which food sensory properties maintain homeostasis. Highlights- A oral dose of astringent flavanols (FLs) immediately fires locus coeruleus-noradrenaline axis - FLs enhance spontaneous locomotion and short-term memory in mice - SAM and HPA axis are also activated by an oral dose of FLs - FLs astringency affects the brain and body, contributing to homeostasis.

synthetic biology↗

Odor source localization behavior of an insect enhanced by intermittent intake strategy

This study investigated odor acquisition strategies to enhance odor plume tracking performance. Efficient odor plume tracking is a crucial ability for organisms, affecting their survival, including for insects with relatively simple nervous systems. Insects can use odor cues to locate food sources or potential mates. Odors released from a source disperse in complex patterns owing to air currents and collisions with objects, making their spread unpredictable. Organisms must therefore engage in active odor acquisition behaviors to effectively gather spatial information from this highly uncertain odor environment. This study focused on odor acquisition via wing flapping in a male silk moth and its relationship with female localization. Given the difficulty of directly intervening in wing flapping, we employed an insect-mounted robotic system to engineer interventions and investigate the relationship between wing-flapping-induced odor acquisition and localization. We found that the difference between air inflow and stoppage in odor attraction was large, and that the odor plume tracking performance was highest at 10 Hz, where odor attraction can be performed at high frequencies. Although constant strong odor acquisition improves localization performance, it increases the likelihood of movement in directions other than that of the odor source. This suggests that periodic wing flapping helps to suppress undesired movements.

animal behavior and cognition↗

PDPN/CLEC-2 axis modulates megakaryocyte subtypes in a hematopoietic stem cell-regulating megakaryocyte-dominant manner

IntroductionMegakaryocytes are classified into several subtypes including LSP1-positive immune-skewed, MYLK4-positive hematopoietic stem cell (HSC)-regulating, and BMAL1-positive platelet-producing megakaryocytes. Podoplanin (PDPN)-expressing stromal cells generate a microenvironment that promotes megakaryopoiesis in the bone marrow. In this context, PDPN interacts with C-type lectin-like receptor-2 (CLEC-2) on megakaryocyte progenitors, which induces megakaryocyte proliferation. However, the megakaryocyte subtypes developed by the regulation of the PDPN/CLEC-2 axis have not yet been elucidated. Materials and MethodsWe established an immortalized bone marrow PDPN-expressing stromal cell line and a PDPN-knockout line (PDPN WT and KO feeder cells, respectively). Bone marrow hematopoietic progenitors were committed to megakaryocytes in co-culture with PDPN WT or KO feeder cells. The number and ploidy of megakaryocytes, resultant platelets, and the polarization of megakaryocyte subtypes were investigated. ResultsThe number of megakaryocytes was significantly increased in the co-culture with PDPN WT feeder cells compared to that with PDPN KO feeder cells. The megakaryocytes on the PDPN WT and KO feeders showed their main ploidy at 16N[~]32N and 8N[~]16N, respectively. The number of platelets decreased in the co-culture with the PDPN WT feeder compared to those in the co-culture with the PDPN KO feeder. Megakaryocyte subtypes were immunocytochemically detected in in vitro differentiated CD41-positive megakaryocytes. For each megakaryocyte subtype, the percentage of MYLK4-positive megakaryocytes significantly increased and the percentage of BMAL1-positive megakaryocytes significantly decreased when co-cultured with the PDPN WT feeder. ConclusionThe PDPN/CLEC-2 axis modulates megakaryocyte subtype differentiation, with a predominance of HSC-regulating megakaryocytes.

cell biology↗

High-coverage genome sequencing of Yayoi and Jomon individuals shed light on prehistoric human population history in East Eurasian

The migration of prehistoric humans led to intriguing interactions and changes in cultural and genetic heritage. In Eurasia, prehistoric migration and population replacement have affected present-day humans. The available high-quality genetic evidence for prehistoric migration in eastern Eurasia, particularly in the Far East, is still limited. We succeeded in obtaining low-contaminant, high-coverage genomes from middle-Yayoi (>46-fold coverage) and Initial Jomon (>67-fold coverage) individuals from mainland Japan. This study demonstrated that the Yayoi individual exhibited a genetic profile distinct from that of the indigenous Jomon population of the Japanese archipelago, suggesting that Yayoi ancestry was connected to the peopling of the Eurasian continent. Our high-coverage genome provides interesting insights into the evolution of copy number polymorphisms related to the dietary styles of ancient Japanese people. The copy number estimates of the amylase gene for the Yayoi individual were comparable to those of present-day East Asians who have diets high in starch. This suggests that the population in the middle Yayoi period may have already adapted to high-starch diets, which may have been related to paddy rice agriculture introduced from the continent. Furthermore, the individual from the initial Jomon period showed high amylase copy numbers comparable to those from modern East Eurasia, including modern Japanese. This suggests that some Jomon people may have consumed a high-starch diet then. The high-coverage whole-genome sequence also revealed differences in the demographic backgrounds of the two ancestral populations during the Yayoi and Jomon periods. Our results shed light on the prehistorical events and origins of related migrations from Eurasia at that time and their genetic background, cultural transformations, and links to modern Japanese people.

genomics↗

Tissue growth associated transition of podoplanin-expressing stromal cells in the postnatal mouse femur

In vertebral long bones, such as the femur, bone formation involves endochondral ossification. Endochondral ossification first occurs in the central region of the fetal diaphysis, the primary ossification center. Podoplanin (PDPN) is a transmembrane mucin-like glycoprotein, and PDPN-positive cells play key roles in various organ/tissue development. In adult mice, osteolineage PDPN-positive cells are associated with femoral microarchitecture. However, specific roles of PDPN in fetal bone development remain unclear. Therefore, in this study, we aimed to investigate the spatiotemporal dynamics and physiological functions of PDPN-positive cells during fetal femur development. In the fetal femur, PDPN-positive cells first emerged in the primitive cortical bone, termed the bone collar, concurrently with primary ossification center initiation. Several PDPN-positive cells in the bone collar migrated to the marrow cavity and populated the metaphyseal trabecular bone. Most PDPN-positive cells in both the bone collar and trabeculae exhibited osteolineage features, such as osterix expression. Pdpn knockout fetuses exhibited abnormal recruitment of osterix-positive cells and mineral deposition in the dorsal bone collar. Overall, our results suggest that PDPN-positive cells constitute a spatially regulated osteolineage population that contributes to coordinated fetal femur development.

cell biology↗

Florigen-producing cells express FPF1-LIKE PROTEIN 1 that accelerates flowering and stem growth in long days with sunlight red/far-red ratio in Arabidopsis

Seasonal changes in spring induce flowering by expressing the florigen, FLOWERING LOCUS T (FT), in Arabidopsis. FT is expressed in unique phloem companion cells with unknown characteristics. The question of which genes are co-expressed with FT and whether they have roles in flowering remains elusive. Through tissue-specific translatome analysis, we discovered that under long-day conditions with the natural sunlight red/far-red ratio, the FT-producing cells express a gene encoding FPF1-LIKE PROTEIN 1 (FLP1). The master FT regulator, CONSTANS (CO), controls FLP1 expression, suggesting FLP1s involvement in the photoperiod pathway. FLP1 promotes early flowering independently of FT, is active in the shoot apical meristem, and induces the expression of SEPALLATA 3 (SEP3), a key E-class homeotic gene. Unlike FT, FLP1 facilitates inflorescence stem elongation. Our cumulative evidence indicates that FLP1 may act as a mobile signal. Thus, FLP1 orchestrates floral initiation together with FT and promotes inflorescence stem elongation during reproductive transitions.

plant biology↗

Basement membrane extract potentiates the endochondral ossification phenotype of bone marrow-derived mesenchymal stem cell-based cartilage organoids

Endochondral ossification is a developmental process in the skeletal system and bone marrow of vertebrates. During endochondral ossification, primitive cartilaginous anlages derived from mesenchymal stem cells (MSCs) undergo vascular invasion and ossification. In vitro regeneration of endochondral ossification is beneficial for research on the skeletal system and bone marrow development as well as their clinical aspects. However, to achieve the regeneration of endochondral ossification, a stem cell-based artificial cartilage (cartilage organoid, Cart-Org) that possesses an endochondral ossification phenotype is required. Here, we modified a conventional 3D culture method to create stem cell-based Cart-Org by mixing it with a basement membrane extract (BME) and further characterized its chondrogenic and ossification properties. BME enlarged and matured the bone marrow MSC-based Cart-Orgs without any shape abnormalities. Histological analysis using Alcian blue staining showed that the production of cartilaginous extracellular matrices was enhanced in Cart-Org treated with BME. Transcriptome analysis using RNA sequencing revealed that BME altered the gene expression pattern of Cart-Org to a dominant chondrogenic state. BME triggered the activation of the SMAD pathway and inhibition of the NK-{kappa}B pathway, which resulted in the upregulation of SOX9, COL2A1, and ACAN in Cart-Org. BME also facilitated the upregulation of genes associated with hypertrophic chondrocytes (IHH, PTH1R, and COL10A1) and ossification (SP7, ALPL, and MMP13). Our findings indicate that BME promotes cartilaginous maturation and further ossification of bone marrow MSC-based Cart-Org, suggesting that Cart-Org treated with BME possesses the phenotype of endochondral ossification. HighlightsO_LIBasement membrane extract (BME) enlarges MSC-based Cart-Org. C_LIO_LIBME activates the SMAD pathway and inhibits the NK-kB pathway of the Cart-Org. C_LIO_LIBME promotes cartilaginous maturation and further ossification of Cart-Org. C_LI

cell biology↗

The Arabidopsis katamari2 Mutant Exhibits a Hypersensitive Seedling Arrest Response at the Phase Transition from Heterotrophic to Autotrophic Growth

Young seedlings use nutrients stored in the seeds to grow and acquire photosynthetic potential. This process, called seedling establishment, involves a developmental phase transition from heterotrophic to autotrophic growth. Some membrane-trafficking mutants of Arabidopsis thaliana (Arabidopsis), such as the katamari2 (kam2) mutant, exhibit growth arrest during seedling development, with a portion of individuals failing to develop true leaves on sucrose-free solid medium. However, the reason for this seedling arrest is unclear. In this study, we show that seedling arrest is a temporal growth arrest response that occurs not only in kam2 but also in wild-type Arabidopsis; however, the threshold for this response is lower in kam2 than in the wild type. A subset of the arrested kam2 seedlings resumed growth after transfer to fresh sucrose-free medium. Growth arrest in kam2 on sucrose-free medium was restored by increasing the gel concentration of the medium or covering the surface of the medium with a perforated plastic sheet. Wild-type Arabidopsis seedlings were also arrested when the gel concentration of sucrose-free medium was reduced. RNA sequencing revealed that transcriptomic changes associated with the rate of seedling establishment were observed as early as 4 days after sowing. Our results suggest that the growth arrest of both kam2 and wild-type seedlings is an adaptive stress response and is not simply caused by the lack of a carbon source in the medium. This study provides a new perspective on an environmental stress response under unfavorable conditions during the phase transition from heterotrophic to autotrophic growth in Arabidopsis.

plant biology↗

Cytosolic heme metabolism by alternative localization of heme oxygenase 1 in plant cells

Heme, an organometallic molecule, is widely engaged in oxygen transport, electron delivery, enzymatic reactions, and signal transduction. Additionally, heme serves as a precursor to phytochromobilin, the chromophore of plant phytochrome. Heme oxygenase (HO) initiates the first committed step in heme metabolism. Our transcription start site-sequencing (TSS-seq) revealed that HO1 in Arabidopsis thaliana and Oryza sativa (rice) has two TSSs, producing long (HO1L) and short (HO1S) transcripts, with or without an intact N-terminal plastid transit peptide. HO1L and HO1S products localize in plastids and the cytosol, respectively. In Arabidopsis, HO1L is prevalent in light-exposed shoots, while HO1S is clearly detected in roots and etiolated seedlings. During de-etiolation and early development, HO1L ratio gradually rises and HO1S ratio decreases. Light perception via phytochrome and cryptochrome elevates HO1L ratio and reduce HO1S ratio through the functioning of HY5 and HYH transcription factors, and the suppression of DET1, E3 ubiquitin ligase COP1, and PIFs transcription factors. As expected, HO1L product was able to complement the HO1-deficient mutant gun2 (hy1), but surprisingly, HO1S expression could also restore the short hypocotyl phenotype and high pigment content, and make the mutant recover from the gun phenotype. This indicates the formation of functional holo-phytochrome within these lines. Our work highlights the presence of a cytosolic pathway for heme metabolism, especially during etiolation and early development. Furthermore, it supports the hypothesis that a mobile heme signal is involved in the mediation of retrograde signaling from the chloroplast. Significance StatementIn this research, through both TSS-seq and CAGE-seq, we discovered that the HO1 (GUN2 or HY1) gene in both Arabidopsis and rice has two TSSs, generating HO1L and HO1S transcripts. We reveal that the HO1 TSS regulation pathway is the same as the light signaling pathway. Significantly, our study identifies that a cytosolic heme metabolism pathway is existent in plant cells.

plant biology↗

Comprehensive Analysis of Peptide-Coding Genes Identifies an LRR-only Microprotein That Regulates Reproductive Organs in Marchantia polymorpha

1In the past two decades, many plant peptides have been found to play crucial roles in various biological events by mediating cell-to-cell communications. However, a large number of small open reading frames (sORFs) or short genes capable of encoding peptides remain uncharacterized. In this study, we examined several candidate genes for peptides conserved between two model plants: Arabidopsis thaliana and Marchantia polymorpha. We examined the expression pattern in M. polymorpha and subcellular localization using a transient assay with Nicotiana benthamiana. We found that one candidate, MpSGF10B, was expressed in meristems, gemma cups, and male reproductive organs called antheridiophores. MpSGF10B has an N-terminal signal peptide followed by two leucine-rich repeat (LRR) domains and was secreted to the extracellular region in N. benthamiana and M. polymorpha. Compared with the wild type, two independent knockout mutants had an increased number of antheridiophores, which emerged from meristems. It was revealed in gene ontology enrichment analysis that MpSGF10B was significantly co-expressed with genes related to cell cycle and development. These results suggest that MpSGF10B may regulate the reproductive development in M. polymorpha. Our research should shed light on the unknown role of LRR-only proteins in land plants.

plant biology↗

A transcription factor module regulating stomatal formation is co-opted for the development of setae in the astomatous liverwort Marchantia polymorpha

The evolution of special types of cells requires the acquisition of new gene regulatory networks controlled by transcription factors (TFs). In stomatous plants, a TF module formed by subfamilies Ia and IIIb basic helix-loop-helix TFs (Ia-IIIb bHLH) regulates stomatal formation; however, how this module evolved during land plant diversification remains unclear. Here, we show that, in the astomatous liverwort Marchantia polymorpha, a Ia-IIIb bHLH module regulates the development of a unique sporophyte tissue, the seta, which is found in mosses and liverworts. The sole Ia bHLH gene, MpSETA, and a IIIb bHLH gene, MpICE2, regulate the cell division and/or differentiation of seta lineage cells. MpSETA can partially replace the stomatal function of Ia bHLH TFs in Arabidopsis thaliana, suggesting that a common regulatory mechanism underlies the setal and stomatal formation. Our findings reveal the co-option of a Ia-IIIb bHLH TF module for regulating cell fate determination and/or cell division of distinct types of cells during land plant evolution.

plant biology↗

Periosteum-derived podoplanin-expressing stromal cells regulate nascent vascularization during epiphyseal marrow development

Bone marrow development and endochondral bone formation occur simultaneously. During endochondral ossification, periosteal vasculatures and stromal progenitors invade the primary avascular cartilaginous anlage; this induces primitive marrow development. We previously determined that bone marrow podoplanin (PDPN)-expressing stromal cells exist in a perivascular microenvironment, and promote megakaryopoiesis and erythropoiesis. In this study, we aimed to examine the involvement of PDPN-expressing stromal cells in the postnatal bone marrow generation. We found that periosteum-derived PDPN-expressing stromal cells regulate vascularization during postnatal epiphyseal marrow development. Our findings suggest that these cells act as pericytes on the primitive vasculature of the nascent marrow. They invade the cartilaginous epiphysis and regulate marrow development and homeostasis by maintaining vascular integrity. To the best of our knowledge, this is the first study to comprehensively examine how PDPN-expressing stromal cells contribute to marrow development and homeostasis.

developmental biology↗