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Harada, R.

Publications and source records attributed to Harada, R..

15 recordsLinked to original sources

GTRspmix: Capturing Heterogeneity of Exchangeabilities Across Sites to Improve Protein Phylogenetics

Site rate and profile mixture models capture the heterogeneity of the amino acid substitution process across sites. However, these models typically use a single matrix of amino acid exchangeabilities and ignore potential heterogeneities of these exchangeabilities across sites. Simply combining multiple exchangeability matrices with rate and profile mixtures leads to a combinatorial explosion of mixture components and a prohibitive increase in free parameters. Here, we introduce GTRspmix, a novel framework that incorporates multiple exchangeability matrices into profile and site rate mixture models while effectively managing model complexity. GTRspmix employs a clustering-based strategy that groups profiles and assigns a distinct exchangeability matrix to each profile cluster. Evaluations using both empirical and simulated datasets demonstrate that GTRspmix fits empirical data significantly better than conventional models, and that overparameterization does not present a problem for sufficiently large alignments. Based on these results, we estimated general-purpose empirical models (SXXpfamCYY series available in IQ-TREE3) from the Pfam database. These general-purpose models not only fit data much better, but they also influence branch length and tree topology estimates, effectively mitigating long-branch attraction artifacts. Because the total number of rate matrices remains manageable, the computational efficiency of the inference is identical to that of conventional profile mixture models (e.g., LG+C60+G4). GTRspmix provides a more realistic and flexible model of protein evolution, offering a robust foundation for the inference of reliable phylogenetic trees.

evolutionary biology↗

Integrated in vivo and transcriptomic analyses of lethal Oropouche virus infection reveal suppression of pathogenic host responses by antiviral therapy

Oropouche virus (OROV) is an emerging arbovirus responsible for large outbreaks of febrile illness in Central and South America, with increasing reports of severe neurological disease and fatal outcomes. Despite its growing public health impact, no approved antiviral therapies or vaccines are currently available. Here, we show that favipiravir, a broad-spectrum nucleoside analogue, robustly suppresses OROV replication and disease in vivo. In a lethal Syrian hamster model, favipiravir treatment provided complete protection against OROV infection, preventing viral dissemination to peripheral organs and the central nervous system, and remained highly effective when administration was initiated after infection. In contrast, insufficient antiviral control resulted in viral neuroinvasion and fatality. To define host responses associated with OROV pathogenesis and their modulation by antiviral therapy, we performed transcriptomic profiling of liver and brain tissues. OROV infection induced interferon-driven inflammatory programs accompanied by marked disruption of metabolic and tissue homeostatic pathways, whereas these transcriptional signatures were largely abrogated by favipiravir treatment. Together, our findings identify favipiravir as a potent antiviral candidate against OROV and provide the first in vivo, tissue-resolved transcriptomic framework of OROV infection, linking effective viral suppression with the prevention of neuroinvasion and pathogenic host responses. These results highlight antiviral intervention as a viable strategy to mitigate OROV-associated disease and mortality.

microbiology↗

A lethal mouse model of Oz virus infection reveals hepatic involvement and enables evaluation of antiviral and vaccine efficacy

Oz virus (OZV), a member of the genus Thogotovirus in the family Orthomyxoviridae, is an emerging tick-borne virus reported in Japan. A fatal human case and seroepidemiological evidence of widespread exposure among wild animals and humans suggest its potential public health significance. However, no animal models suitable for pathogenic studies or evaluation of countermeasures are available for OZV. Here, we have established a lethal mouse model of OZV infection using cell-adapted virus and mice lacking type I interferon signaling (B6 Ifnar1 KO mice). OZV infection resulted in 100% mortality and was characterized by robust viral replication in the liver and spleen, severe hepatitis, and acute liver injury. Using this model, we also demonstrated that oral administration of T-705, an antiviral drug widely used against RNA viruses, as well as immunization with an inactivated whole virus particle vaccine, protected B6 Ifnar1 KO mice from lethal OZV infection by mitigating the acute hepatitis. The present study provides critical insights into OZV pathogenesis and establishes a practical in vivo platform for the development of countermeasures against OZV infection. Significance statementEmerging tick-borne viruses pose a growing public health concern, yet progress in understanding their pathogenesis and in developing countermeasures is often limited by the lack of relevant animal models. Oz virus (OZV), a recently identified thogotovirus associated with a fatal human case, exemplifies this challenge. Here, we establish a lethal mouse model of OZV infection that reveals pronounced hepatic involvement as a central pathological feature. Using this model, we demonstrate effective protection by both an antiviral drug and an inactivated vaccine against lethal OZV challenge. This study provides a practical in vivo platform for investigating OZV pathogenesis and for accelerating the development of medical countermeasures against this emerging tick-borne virus.

microbiology↗

Three-dimensional and molecular brain atlas of the hagfish reveals the evolutionary origin and early diversification of the vertebrate brain

Hagfish and lampreys form the clade cyclostomes, the earliest branching lineage of the vertebrates. Although hagfish are generally assumed to have experienced intense modifications, its brain development shows certain ancestral patterns that are secondarily lost in the lamprey. Therefore, hagfish provides key insights into the evolutionary origin and early diversification of the vertebrate brain. However, neuroanatomical information of the hagfish brain is substantially limited today due to its sources based on old-fashioned methodology alone. Here, we provide a detailed brain atlas of the adult hagfish, based on three-dimensional reconstruction and expression analysis of genetic markers for major neuronal transmitters. Through our analyses, we found the following key characteristics. First, the dopaminergic system appears to be distributed more broadly than previously thought in the hagfish brain, implying that dopamine is involved in various neural functions. Second, the previously suggested "primordial cerebellum" area in hagfish shows notable affinity to the cerebellum-like octavolateral structures of the jawed vertebrate hindbrain. Last, the gene expression profile suggests a hippocampus-equivalent brain region in hagfish, that is, the ventrolateral subnucleus of the central prosencephalic complex (NCvl). This study highlights conserved and diversified neuroarchitecture of the hagfish brain, providing a pivotal reference for further studies.

evolutionary biology↗

Genome-wide and gene expression analyses of the hagfish olfactory receptors illuminate lineage-specific diversification of olfaction in basal vertebrates

Olfactory receptors (ORs) of vertebrates have undergone remarkable diversification, but the ancestral repertoire remains incompletely understood. Here, we present a comprehensive genome-wide survey and gene expression analyses of OR genes in hagfish, a cyclostome group that typically inhabits the deep sea with a well-developed olfactory system. We identified 48 main olfactory receptors (mORs), two vomeronasal type 1 receptors (V1Rs), 135 typical vomeronasal type 2 receptors (V2Rs), and no trace amine-associated receptors (TAARs) in the inshore hagfish (Eptatretus burgeri). Notably, V2Rs were highly clustered on a single scaffold, suggesting tandem gene duplications as the main mechanism of diversification. Transcriptome analysis confirmed that most of these OR genes are expressed predominantly in the olfactory organ. Using in situ hybridization, we further confirmed that at least some of these genes are expressed in olfactory receptor cells. These gene expression analyses support the role of these genes as functional ORs. Our findings challenge the prevailing view that typical olfactory V2Rs originated in gnathostomes and instead suggest that they were present in the common ancestor of all vertebrates but were lost in lampreys. The extensive diversification of V2Rs in hagfish likely reflects adaptations to life in low-light and chemically complex environments. This study highlights the evolution of vertebrate sensory systems in a mosaic manner, underscoring the importance of hagfish as a key animal for reconstructing the evolutionary history of early vertebrates. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/666228v2_ufig1.gif" ALT="Figure 1"> View larger version (42K): org.highwire.dtl.DTLVardef@ba336borg.highwire.dtl.DTLVardef@85b62corg.highwire.dtl.DTLVardef@7b23ccorg.highwire.dtl.DTLVardef@6b2b74_HPS_FORMAT_FIGEXP M_FIG C_FIG Highlights- Genomic and expression analyses reveal hagfish olfactory receptor repertoire. - V2Rs are extensively diversified in hagfish. - The olfactory function of V2Rs is traced back to the common ancestor of vertebrates. - The olfactory receptor repertoires of basal vertebrates underwent mosaic evolution.

evolutionary biology↗

Phylogenetic dissection provides insights into the incongruity in the tree of Archaeplastida between the analyses of nucleus- and plastid-encoded proteins.

Archaeplastida is defined as a taxonomic assemblage comprising three sub-clades, namely Chloroplastida, Glaucophyta, and Rhodophyta plus two non-photosynthetic lineages sister to Rhodophyta (the latter three lineages collectively termed "Rhodozoa" here). The members of Archaeplastida are the descendants of the eukaryote that took up and transformed a cyanobacterial endosymbiont into a primary plastid. Recent phylogenetic analyses of multiple proteins (phylogenomic analyses) stably recovered the monophyly of Archaeplastida, but uncertainty remains in the relationship among the three sub-clades in this assemblage. The phylogenomic analyses of nucleus-encoded proteins (nuc-proteins) grouped Chloroplastida and Glaucophyta together, excluding Rhodozoa in the Archaeplastida clade, albeit the union of Chloroplastida and Rhodophyta was often inferred from the phylogenomic analyses of plastid-encoded proteins (pld-proteins). In this study, we challenged the previously recognized but as-yet-explicitly addressed issue in the tree of Archaeplastida (ToA). The detailed analyses of the nuc-protein and pld-protein supermatrices revealed that taxon sampling can invoke different types of phylogenetic artifacts into the inferences from both supermatrices examined here. In the end, we propose a working hypothesis for the ToA and provide future perspectives toward resolving the ToA.

evolutionary biology↗

A cellular entity retaining only its replicative core: Hidden archaeal lineage with an ultra-reduced genome

Defining the minimal genetic requirements for cellular life remains a fundamental question in biology. Genomic exploration continually reveals novel microbial lineages, often exhibiting extreme genome reduction, particularly within symbiotic relationships. Here, we report the discovery of Candidatus Sukunaarchaeum mirabile, a novel archaeon with an unprecedentedly small genome of only 238 kbp --less than half the size of the smallest previously known archaeal genome-- from a dinoflagellate-associated microbial community. Phylogenetic analyses place Sukunaarchaeum as a deeply branching lineage within the tree of Archaea, representing a novel major branch distinct from established phyla. Environmental sequence data indicate that sequences closely related to Sukunaarchaeum form a diverse and previously overlooked clade in microbial surveys. Its genome is profoundly stripped-down, lacking virtually all recognizable metabolic pathways, and primarily encoding the machinery for its replicative core: DNA replication, transcription, and translation. This suggests an unprecedented level of metabolic dependence on a host, a condition that challenges the functional distinctions between minimal cellular life and viruses. The discovery of Sukunaarchaeum pushes the conventional boundaries of cellular life and highlights the vast unexplored biological novelty within microbial interactions, suggesting that further exploration of symbiotic systems may reveal even more extraordinary life forms, reshaping our understanding of cellular evolution. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=123 SRC="FIGDIR/small/651781v1_ufig1.gif" ALT="Figure 1"> View larger version (41K): org.highwire.dtl.DTLVardef@f5fbd6org.highwire.dtl.DTLVardef@168bee5org.highwire.dtl.DTLVardef@1ccb02forg.highwire.dtl.DTLVardef@14315fa_HPS_FORMAT_FIGEXP M_FIG C_FIG

evolutionary biology↗

Complete mitochondrial genomes of ancyromonads provide clues for the gene content and genome structures of ancestral mitochondria.

Mitochondria of eukaryotic cells are direct descendants of an endosymbiotic bacterium related to Alphaproteobacteria. These organelles retain their own genomes, which are highly reduced and divergent when compared to those of their bacterial relatives. To better understand the trajectory of mitochondrial genome evolution from the last eukaryotic common ancestor (LECA) to extant species, mitochondrial genome sequences from phylogenetically diverse lineages of eukaryotes - particularly protists - are essential. For this reason, we focused on the mitochondrial genomes of Ancyromonadida, an independent and understudied protist lineage in the eukaryote tree of life. Here we report the mitochondrial genomes from three Ancyromonadida: Ancyromonas sigmoides, Nutomonas longa, and Fabomonas tropica. Our analyses reveal that these mitochondrial genomes are circularly mapping molecules that feature protein-coding inverted repeats. This inverted repeat structure has been observed in other mitochondrial genomes but is patchily distributed over the tree of eukaryotes. Ancyromonad mitochondrial genomes possess several protein-coding genes which have not been detected from any other mitochondrial genomes of eukaryotes sequenced to date, thereby extending the known mitochondrial gene repertoire of ancestral eukaryotes, including LECA. These findings significantly expand our understanding of mitochondrial genome diversity across eukaryotes, shedding light on the early phases of mitochondrial genome evolution in eukaryotes.

evolutionary biology↗

Glissandra oviformis n. sp.: a novel predatory flagellate illuminates the character evolution within the eukaryotic clade CRuMs

Culturing protists offers a powerful approach to exploring eukaryotic diversity, especially for deep-branching lineages. In this study, we cultured and described a novel protist species, named Glissan-dra oviformis n sp., within the poorly studied and unclassified genus Glissandra. While an SSU rDNA gene phylogeny failed to resolve its phylogenetic placement in the eukaryotic tree, a phylogenomic analysis of 340 proteins indicated G. oviformis as a deep-branching member of the CRuMs clade. Prior to this study, this clade consisted of diverse het-erotrophic amoeba and flagellates and lacks clear synapomorphies. Ultrastructural observations re-vealed that G. oviformis shares the characteristics with some CRuMs members, including the pellicle underlying the plasma membrane and an internal sleeve surrounding the central pair of the axoneme at the flagellar transitional region. Our findings suggest potential shared characteristics and synapomorphies for CRuMs and contribute to a deeper understanding of the character evolution within CRuMs.

evolutionary biology↗

Drastic genome reduction driven by parasitic lifestyle: Two complete genomes of endosymbiotic bacteria possibly hosted by a dinoflagellate

Bacteria with endosymbiotic lifestyles often exhibit dramatic genome reduction. While the reduction of genomes in intracellular symbionts of animals, including parasitic bacteria, has been extensively studied, less is known about such bacteria associated with single-celled eukaryotes. Here, we report the genomes of two novel gammaproteobacterial lineages, RS3 and XS4, identified as putative parasitic endosymbionts of the dinoflagellate Citharistes regius. Phylogenetic analyses suggest that RS3 and XS4 belong to the family Fastidiosibacteraceae within the order Beggiatoales, forming independent lineages therein. The genomes of RS3 and XS4 are 529 Kbp and 436 Kbp in size, respectively, revealing drastic genome reduction compared to related bacterial genomes. XS4, which has a particularly reduced genome with low GC content, uses an alternative genetic code, in which UGA assigned tryptophan. The small genomes of RS3 and XS4 encode a limited number of proteins, retaining only approximately 20% of the predicted ancestral proteome. Metabolic reconstruction suggests that RS3 and XS4 are energy parasites, heavily dependent on their host for essential metabolites. Furthermore, we found that the ancestor of both genomes likely acquired an ADP:ATP antiporter gene via horizontal gene transfer, an event that may have enabled their evolution as energy parasites by facilitating the acquisition of ATP from their host. These observations on novel bacteria with highly reduced genomes expand our understanding of the phylogenetic and genomic diversity of endosymbiotic bacteria in protists.

evolutionary biology↗

Harnessing Synaptic Vesicle Release and Recycling Mechanism for Molecule Delivery to Neurons

Neurodegenerative clinical trials often fail due to insufficient drug doses in reaching targeted cells and the unintended delivery to non-targeted cells. This study demonstrates an alternative neuron-selective drug delivery system, which utilizes the synaptic vesicle release and recycling mechanism (SVRM) by antibody shuttles targeting synaptic vesicle transmembrane proteins for molecule delivery. Using Synaptotagmin-2 (SYT2), we exemplify that intravenously administered anti-SYT2 antibodies localize to neuromuscular junctions, undergo uptake, and retrograde transport to ChAT-positive motor neurons (MNs) in the spinal cord and brainstem. The delivery of anti-microtubule agent and Malat1 gapmer antisense oligonucleotide to MNs with anti-SYT2 antibodies induces axon degeneration and reduction of Malat1 RNA expression, respectively. This approach circumvents the blood-spinal cord barrier, enabling selective delivery of therapeutic molecules to neurons while minimizing effects in non-targeted cells. Thus harnessing SVRM presents a promising strategy for enhancing drug concentrations in neurons and improving treatment efficacy for neurodegenerative diseases.

neuroscience↗

Encyclopaedia of DNA polymerases localized in organelles: Contribution of genes from diverse bacteria including the proto-mitochondrion.

DNA polymerases (DNAPs) synthesize DNA from deoxyribonucleotides in a semi-conservative manner and serve as the core of DNA replication and repair machineries. In eukaryotic cells, there are two genome-containing organelles, mitochondria and plastids, that were derived from an -proteobacterium and a cyanobacterium, respectively. Except for rare cases of genome-lacking mitochondria and plastids, both organelles must be served by nucleus-encoded DNAPs that localize and work in them to maintain their genomes. The evolution of organellar DNAPs has yet to be fully understood because of two unsettled issues. First, the diversity of organellar DNAPs has not been elucidated in the full spectrum of eukaryotes. Second, it is unclear when the DNAPs that were used originally in the endosymbiotic bacteria giving rise to mitochondria and plastids were discarded, as the organellar DNAPs known to date show no phylogenetic affinity to those of the extant -proteobacteria or cyanobacteria. In this study, we identified from diverse eukaryotes 134 family A DNAP sequences, which were classified into 10 novel types, and explored their evolutionary origins. The subcellular localizations of selected DNAPs were further examined experimentally. The results presented here suggest that the diversity of organellar DNAPs has been shaped by multiple transfers of the PolI gene from phylogenetically broad bacteria, and their occurrence in eukaryotes was additionally impacted by secondary plastid endosymbioses. Finally, we propose that the last eukaryotic common ancestor may have possessed two mitochondrial DNAPs, POP and a candidate of the direct descendant of the proto-mitochondrial DNAP, rdxPolA, identified in this study.

evolutionary biology↗

Autism-associated mutation in Hevin/Sparcl1 induces endoplasmic reticulum stress through structural instability

Hevin is a secreted extracellular matrix protein that is encoded by SPARCL1 gene. Recent studies show that Hevin plays an important role in regulating synaptogenesis and synaptic plasticity. Mutations in SPARCL1 gene increase the risk of autism spectrum disorder (ASD). However, the molecular basis of how mutations in SPARCL1 increase the risk of ASD has not been fully understood. In this study, we show that one of SPARCL1 mutations associated with ASD impairs normal Hevin secretion. We identified Hevin mutants lacking the EF-hand motif through analyzing ASD-related mice with vulnerable spliceosome functions. Hevin deletion mutants accumulate in the ER, leading to the activation of unfolded protein responses. We also found that a single amino acid substitution of Trp647 with Arg in the EF-hand motif associated with a familial case of ASD causes a similar phenotype with the EF-hand deletion mutant. Importantly, molecular dynamics (MD) simulation revealed that this single amino acid substitution triggers exposure of hydrophobic amino acid to the surface, increasing the binding of Hevin with a molecular chaperon, BIP. Taken together, these data suggest that the integrity of EF-hand motif in Hevin is crucial for proper folding and ASD-related mutation impairs an export of Hevin from the endoplasmic reticulum (ER). Our data provide a novel mechanism linking a point mutation in SPARCL1 gene to the molecular and cellular characteristics involved in ASD.

molecular biology↗

Phage origin of mitochondrion-localized family A DNA polymerases in kinetoplastids and diplonemids

Mitochondria retain their own genomes as other bacterial endosymbiont-derived organelles. Nevertheless, no protein for DNA replication and repair is encoded in any mitochondrial genomes (mtDNAs) assessed to date, suggesting the nucleus primarily governs the maintenance of mtDNA. As the proteins of diverse evolutionary origins occupy a large proportion of the current mitochondrial proteomes, we anticipate finding the same evolutionary trend in the nucleus-encoded machinery for mtDNA maintenance. Indeed, none of the DNA polymerases (DNAPs) in the mitochondrial endosymbiont, a putative -proteobacterium, seemingly had been inherited by their descendants (mitochondria), as none of the known types of mitochondrion-localized DNAP showed a specific affinity to the -proteobacterial DNAPs. Nevertheless, we currently have no concrete idea of how and when the known types of mitochondrion-localized DNAPs emerged. We here explored the origins of mitochondrion-localized DNAPs after the improvement of the samplings of DNAPs from bacteria and phages/viruses. Past studies revealed that a set of mitochondrion-localized DNAPs in kinetoplastids and diplonemids, namely PolIB, PolIC, PolID, PolI-Perk1/2, and PolI-dipl (henceforth designated collectively as "PolIBCD+") have emerged from a single DNAP. In this study, we recovered an intimate connection between PolIBCD+ and the DNAPs found in a particular group of phages. Thus, the common ancestor of kinetoplastids and diplonemids most likely converted a laterally acquired phage DNAP into a mitochondrion-localized DNAP that was ancestral to PolIBCD+. The phage origin of PolIBCD+ hints at a potentially large contribution of proteins acquired via non-vertical processes to the machinery for mtDNA maintenance in kinetoplastids and diplonemids.

evolutionary biology↗

Barthelonids represent a deep-branching Metamonad clade with mitochondrion-related organelles generating no ATP.

We here report the phylogenetic position of barthelonids, small anaerobic flagellates previously examined using light microscopy alone. Barthelona spp. were isolated from geographically distinct regions and we established five laboratory strains. Transcriptomic data generated from one Barthelona strain (PAP020) was used for large-scale, multi-gene phylogenetic (phylogenomic) analyses. Our analyses robustly placed strain PAP020 at the base of the Fornicata clade, indicating that barthelonids represent a deep-branching Metamonad clade. Considering the anaerobic/microaerophilic nature of barthelonids and preliminary electron microscopy observations on strain PAP020, we suspected that barthelonids possess functionally and structurally reduced mitochondria (i.e. mitochondrion-related organelles or MROs). The metabolic pathways localized in the MRO of strain PAP020 were predicted based on its transcriptomic data and compared with those in the MROs of fornicates. Strain PAP020 is most likely incapable of generating ATP in the MRO, as no mitochondrial/MRO enzymes involved in substrate-level phosphorylation were detected. Instead, we detected the putative cytosolic ATP-generating enzyme (acetyl-CoA synthetase), suggesting that strain PAP020 depends on ATP generated in the cytosol. We propose two separate losses of substrate-level phosphorylation from the MRO in the clade containing barthelonids and (other) fornicates.

evolutionary biology↗