Biology subjects

Mizutani, T.

Publications and source records attributed to Mizutani, T..

11 recordsLinked to original sources

bioRxiv2025

MHC Class I-like Gene in Squirrel Monkey Cytomegalovirus and the Evolution of the Virus

Homology searches between viral and human peptides revealed that the S9 protein of squirrel monkey cytomegalovirus (Saimiriine herpesvirus 4 strain [SBHV4]) shares 60.6% amino acid identity across 86.3% of full-length human HLA-A. This represents the most extensive and highest degree of homology reported to date between a viral protein and an MHC class I molecule. Phylogenetic analysis of MHC class I-like DNA sequences from primate HLA-A orthologs and SBHV4-S9 gene suggested that SBHV4-S9 was likely acquired via LINE-1-mediated horizontal gene transfer from the African Old World monkey Piliocolobus tephrosceles or a close ancestor 1.63-6.02 million years ago. SBHV4-S9 appeared to have undergone purifying selection following divergence from P. tephrosceles. Furthermore, phylogenetic analysis of the SBHV4-S1 gene and primate SLAMF6-like genes suggested that this gene was acquired after divergence from a close ancestor of the New World monkeys Saimiri oerstedii and Saimiri sciureus. Comprehensive evaluation of the similarity between eight SBHV4 genes (including S1 and S9) and their respective homologs in 241 primate species, assessed via PCA-syncmer-based Wasserstein distances, revealed that the overall relatedness between SBHV4 and African Cercopithecus ascanius, Cercopithecus pogonias, and Allenopithecus nigroviridis, respectively was significantly greater (i.e., exhibited shorter Wasserstein distances) in each case than that observed with all examined members of the genus Saimiri and Piliocolobus tephrosceles. Collectively, these findings indicate that SBHV4, although isolated from Saimiri sciureus in South America, appears to have a historical connection with Old World monkeys in Africa, despite the typically high host specificity of cytomegaloviruses.

microbiology↗

bioRxiv2024

Sensitivity to Vγ9Vδ2TCR T cells is imprinted after single mutations during early oncogenesis

V{gamma}9V{delta}2T cells have the unique ability to recognize a broad range of malignant transformed cells. The tumor targeting event involving BTN2A1 and BTN3A1 dimers on the tumor cell surface is critical, leading to full activation of the TCR. Although the molecular mechanisms governing TCR engagement and T cell activation are well-characterized, the role of V{gamma}9V{delta}2 T cells in cancer immune surveillance remains to be fully elucidated, particularly the mechanisms that enable these cells to discriminate between healthy and malignant cells at an early stage of malignant transformation. We employed two independent, genetically engineered step-wise mutagenesis models of human colorectal and breast cancer that mimic the transformation steps leading to tumor formation. We demonstrate that various single oncogenic mutations introduced into healthy organoids or cells, are sufficient to upregulate surface expressed BTN2A1 and enable V{gamma}9V{delta}2 TCR binding to tumor cells. However, full activation of T cells through a V{gamma}9V{delta}2TCR required additional subsequent phosphorylation of juxtamembrane (JTM) amino acids of BTN3A1, leading to the activating heterodimerization of BTN2A1 and 3A1. Using a protein interactome mapping pipeline, we identified PHLDB2, SYNJ2 and CARMIL1 as key players in controlling these delicate dual surface dynamics of BTN2A1 and 3A1 during early transformation. This mode of action allowed V{gamma}9V{delta}2TCR T cells to control tumors in vitro and in vivo, emphasizing the crucial role of these molecules from early mutagenesis, to advanced cancer stages, and highlighting the therapeutic potential of a V{gamma}9V{delta}2TCR.

cancer biology↗

bioRxiv2024

Entomopoxvirus-like long DNA sequences in human centromeric and peri-centromeric regions

Mammalian genomes harbor a range of endogenous virus-like sequences, although these sequences are typically affected by mutations and deletions, and no examples have been reported of long, conserved regions extending tens of kilobases or more. In this study, a comprehensive similarity search against the complete human T2T genome (hT2T) using high-quality viral genome data was performed. Among nearly continuous viral sequences, the longest identified was Entomopoxvirus, spanning 140 kb with a similarity of over 57%. Notably, Entomopoxvirus-like sequences stood out within the human genome, with a total of 2.42 Mb exhibiting similarity to human sequences. Seven Entomopoxvirus species were fragmented into regions less than 40 kb each, with nearly all of these segments scattered throughout the human genome. Surprisingly, nearly all of these sequences were localized within the hsat1A regions of human centromeric and pericentromeric areas. On chromosome 3, for example, all sequences were confined within the centromere, and extensive transcription was observed across these regions. While the lack of fully sequenced genomes with solid annotations for other eukaryotic species limits our ability to infer the phylogenetic significance of these regions, our findings reveal remarkably long virus-like sequences within centromeric and related regions--as integral components of eukaryotic chromosomes.

bioinformatics↗

bioRxiv2024

Giant extrachromosomal element "Inocle" potentially expands the adaptive capacity of the human oral microbiome

Survival strategy of bacteria is expanded by extrachromosomal elements (ECEs). However, their genetic diversity and functional roles for adaptability are largely unknown. Here, we discovered a novel family of intracellular ECEs using 56 saliva samples by developing an efficient microbial DNA extraction method coupled with long-read metagenomics assembly. Even though this ECE family was not hitherto unidentified, our global prevalence analysis using 476 salivary metagenomic datasets elucidated that these ECEs reside in 74% of the population. These ECEs, which we named, "Inocles", are giant plasmid-like circular genomic elements of 395 kb in length, having Streptococcus as a host bacterium. Inocles encode a series of genes that contribute to intracellular stress tolerance, such as oxidative stress and DNA damage, and cell wall biosynthesis and modification involved in the interactions with oral epithelial cells. Moreover, Inocles exhibited significant positive correlations with immune cells and proteins responding to microbial infection in peripheral blood. Intriguingly, we examined and found their marked reductions among 68 patients of head and neck cancers and colorectal cancers, suggesting its potential usage for a novel biomarker of gastrointestinal cancers. Our results suggest that Inocles potentially boost the adaptive capacity of host bacteria against various stressors in the oral environment.

genomics↗

bioRxiv2022

Centrosome Loss And Cell Proliferation Defects Underlie Developmental Failure In Haploid Zebrafish Larvae

Haploid larvae in non-mammalian vertebrates are lethal with characteristic organ growth retardation collectively called "haploid syndrome." In contrast to mammals whose haploid intolerance is attributed to imprinting misregulation, the cellular principle of haploidy-linked defects in non-mammalian vertebrates remains unknown. Here, we investigated cellular defects that disrupt the ontogeny of gynogenetic haploid zebrafish larvae. Unlike diploid control, haploid larvae manifested unscheduled cell death at the organogenesis stage, attributed to haploidy-linked p53 upregulation. Moreover, we found that haploid larvae specifically suffered the gradual aggravation of mitotic spindle monopolarization during 1-3 days post fertilization, causing spindle assembly checkpoint-mediated mitotic arrest throughout the entire body. High-resolution imaging revealed that this mitotic defect accompanied the haploidy-linked centrosome loss occurring concomitantly with the gradual decrease in larval cell size. Either resolution of mitotic arrest or depletion of p53 partially improved organ growth in haploid larvae. Based on these results, we propose that haploidy-linked mitotic defects and cell death are parts of critical cellular causes shared among vertebrates that limit the larval growth in the haploid state, contributing to an evolutionary constraint on allowable ploidy status in the vertebrate life cycle.

developmental biology↗

bioRxiv2022

Distinct Effects of the Male-Killing Bacteria Wolbachia and Spiroplasma and a Partiti-Like Virus in the Tea Pest Moth, Homona magnanima

Male killing, the phenomenon of male death during development, is considered to be one of the advantageous strategies exerted by maternally transmitted microbes. Male killing has attracted interest in the fields of evolutionary biology and ecology for decades; however, little is known about its mechanism and origin. Here, we characterized and compared the effects of three distinct male killers, Wolbachia (Alphaproteobacteria), Spiroplasma (Mollicutes), and Osugoroshi virus (OGV) (Partitiviridae) in the tea pest moth Homona magnanima (Lepidoptera, Tortricidae). Regardless of the genetic sex (male: ZZ; female: ZW), female specific splice variants of the doublesex gene (dsx), a downstream regulator of the sex-determining gene cascade, was expressed in H. magnanima harbored either male-killing Wolbachia or Spiroplasma. However, OGV and non-male-killing Wolbachia did not alter dsx splicing. RNA sequencing and quantitative PCR assays demonstrated that male-killing Wolbachia impaired the hosts dosage compensation system by altering the global gene expression of the Z chromosome (corresponding to Bombyx mori chromosome 1 and 15) in males, whereas Spiroplasma did not affect dosage compensation. In contrast, the partiti-like virus OGVs did not affect sex-determination cascades or dosage compensation systems. Besides, male killers distinctly altered host gene expression and metabolomes associated with physiology, morphology, and diverse metabolic pathways. Moreover, Wolbachia and Spiroplasma infections triggered abnormal apoptosis only in male embryos. These findings suggest that distantly related microbes employ distinct machineries to kill identical host males, which have been acquired through independent evolutionary processes. ImportanceMale-killing caused by diverse microbes has attracted substantial attention. However, it remains unclear how such male killers have evolved similar phenotypes, in part because male-killing mechanisms have been studied using different insect models. Here, by comparing three phylogenetically distinct male killers, Wolbachia, Spiroplasma, and a partiti-like virus, in an identical host, we provide evidence that microbes can affect male viability through distinct machinery, demonstrating distinct evolutionary scenarios for microbes to acquire make-killing ability. These findings provide insight into new directions for studying microbe-host interactions.

evolutionary biology↗

bioRxiv2021

Development of PRPK Directed Phthalimides

Immunomodulatory drugs (IMiDs) thalidomide, lenalidomide, and pomalidomide (Pom) bind to cereblon (CRBN) and trigger proteasomal degradation of neo-substrates IKZF1/3 leading to multiple myeloma (MM) cell apoptosis. Pomalidomide (Pom) also binds albeit weakly to p53-related protein kinase (PRPK, aka TP53RK), an understudied kinase reported to be associated with poor prognosis in MM patients. Here, we developed a series of IMiDs based on Pom and conducted a structure-activity relationship (SAR) study to identify a potent and selective PRPK binder. Structural analysis showed that IMiDs bind PRPK in a fundamentally different way from CRBN, and suggested specific derivatization to improve affinity. We employed a structure-guided strategy to develop compound TXM-02-118, which exhibited nanomolar affinityfor PRPK in binding assays, and showed high selectivity for PRPK over CRBN. Overall, the work represents an initial effort to develop tool compounds for studying PRPK. Moreover, it illustrates how a single class of molecules can use different recognition elements to bind diverse targets using fundamentally different binding poses. This has broad implications for chemical probe and lead compound selectivity profiling, and argues for more wide-spread use of global proteomics or similar methodologies.

biochemistry↗

bioRxiv2021

Intensive Single Cell Analysis Reveals Immune Cell Diversity among Healthy Individuals

It is believed that immune responses are different between individuals and at different times. In addition, personal health histories and unique environmental conditions should collectively determine the present state of immune cells. However, the cellular and molecular system mechanisms underlying such heterogeneity remain largely elusive. In this study, we conducted a systematic time-lapse single-cell analysis, using 171 single-cell libraries and 30 mass cytometry datasets intensively for seven healthy individuals. We found substantial diversity in immune cell populations and their gene expression patterns between different individuals. These patterns showed daily fluctuations even within the same individual spending a usual life. Similar diversities were also observed for the T cell receptor and B cell receptor repertoires. Detailed immune cell profiles at healthy statuses should give an essential background information to understand their immune responses, when the individual is exposed to various environmental conditions. To demonstrate this idea, we conducted the similar analysis for the same individuals on the vaccination of Influenza and SARS-CoV-2, since the date and the dose of the antigens are well-defined in these cases. In fact, we found that the distinct responses to vaccines between individuals, althougth key responses are common. Single cell immune cell profile data should make fundamental data resource to understand variable immune responses, which are unique to each individual.

genomics↗

bioRxiv2021

KSR1-and ERK-dependent Translational Regulation of the Epithelial-to-Mesenchymal Transition

The epithelial-to-mesenchymal transition (EMT) is considered a transcriptional process that induces a switch in cells from a polarized state to a migratory phenotype. Here we show that KSR1 and ERK promote EMT through the preferential translation of Epithelial-Stromal Interaction 1 (EPSTI1), which is required to induce the switch from E-to N-cadherin and coordinate migratory and invasive behavior. EPSTI1 is overexpressed in human colorectal cancer (CRC) cells. Disruption of KSR1 or EPSTI1 significantly impairs cell migration and invasion in vitro, and reverses EMT, in part, by decreasing the expression of N-cadherin and the transcriptional repressors of E-cadherin expression, ZEB1 and Slug. In CRC cells lacking KSR1, ectopic EPSTI1 expression restored the E-to N-cadherin switch, migration, invasion, and anchorage-independent growth. KSR1-dependent induction of EMT via selective translation of mRNAs reveals its underappreciated role in remodeling the translational landscape of CRC cells to promote their migratory and invasive behavior.

cancer biology↗

bioRxiv2019

A novel defective recombinant porcine enterovirus G virus carrying a porcine torovirus papain-like cysteine protease gene and a putative anti-apoptosis gene in place of viral structural protein genes

Enterovirus G (EV-G) belongs to the family of Picornaviridae. Two types of recombinant porcine EV-Gs carrying papain-like cysteine protease (PLCP) gene of porcine torovirus, a virus in Coronaviridae, are reported. Type 1 recombinant EV-Gs are detected in pig feces in Japan, USA, and Belgium and carry the PLPC gene at the junction site of 2C/3A genes, while PLPC gene replaces the viral structural genes in type 2 recombinant EV-G detected in pig feces in a Chinese farm. We identified a novel type 2 recombinant EV-G carrying the PLCP gene with flanking sequences in place of the viral structural genes in pig feces in Japan. The ~0.3 kb-long upstream flanking sequence had no sequence homology with any proteins deposited in GenBank, while the downstream ~0.9 kb-long flanking sequence included a domain having high amino acid sequence homology with a baculoviral inhibitor of apoptosis repeat superfamily. The pig feces, where the novel type 2 recombinant EV-G was detected, also carried type 1 recombinant EV-G. Although the phylogenetic analysis suggested that these two recombinant EV-Gs have independently evolved, type 1 recombinant EV-G might have served as a helper virus by providing viral structural proteins for dissemination of the type 2 recombinant EV-G.

microbiology↗

bioRxiv2018

doublesex regulates sexually dimorphic beetle horn formation by integrating spatial and temporal developmental contexts in the Japanese rhinoceros beetle Trypoxylus dichotomus

AbstractMany scarab beetles have sexually dimorphic exaggerated horns that are an evolutionary novelty. Since the shape, number, size, and location of horns are highly diverged within Scarabaeidae, beetle horns are an attractive model for studying the evolution of sexually dimorphic and novel traits. In beetles including the Japanese rhinoceros beetle Trypoxylus dichotomus, the sex determination gene doublesex (dsx) plays a crucial role in sexually dimorphic horn formation during larval-pupal development. However, knowledge of when and how dsx drives the gene regulatory network (GRN) for horn formation to form sexually dimorphic horns during development remains elusive. To address this issue, we identified a Trypoxylus-ortholog of the sex determination gene, transformer (tra), that regulates sex-specific splicing of the dsx pre-mRNA, and whose loss of function results in sex transformation. By knocking down tra function at multiple developmental timepoints during larval-pupal development, we estimated the onset when the sex-specific GRN for horn formation is driven. In addition, we also revealed that dsx regulates different aspects of morphogenetic activities during the prepupal and pupal developmental stages to form appropriate morphologies of pupal head and thoracic horn primordia as well as those of adult horns. Based on these findings, we discuss the evolutionary developmental background of sexually dimorphic trait growth in horned beetles.\n\nAuthor Summary:\n\nBeetle horns are highly enriched in a particular family Scarabaeidae, although the shape, size and number of horns are diversified within the group. In addition, many scarab beetle horns are sexually dimorphic. It has been questioned how a particular group of beetles has originated and diversified evolutionary novel horns. Here we found the exact time when morphological sexual dimorphism of horn primordia appeared, estimated the onset of the developmental program for sexually dimorphic horn formation driven by Doublesex, and revealed that Doublesex regulates different aspects of cell activities of horn primordia depending on the spatiotemporal contexts. Our study provides our understanding regarding regulatory shifts in these mechanisms during the evolution of sexually dimorphic traits in horned beetles.

developmental biology↗