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Kuraku, S.

Publications and source records attributed to Kuraku, S..

At least 19 recordsLinked to original sources

Near-complete genome assembly of the African turquoise killifish enables locus-level profiling of transposable element expression

Tandem repeats (TRs) and transposable elements (TEs) occupy substantial fractions of eukaryotic genomes, yet how repeat detection programs affect downstream expression analyses remains poorly explored. Here, we present an improved chromosome-scale assembly of the African turquoise killifish, Nothobranchius furzeri. The assembly matches experimentally estimated genome size and chromosome number. It resolves highly expanded TRs that occupy up to 28.0% of the whole genome. Using this assembly and other complete vertebrate genome assemblies, we compared TR and TE-derived regions across combinations of TR detection programs and TE consensus library construction workflows under a standardized TE classification scheme. Choices in TR detection reshaped the apparent TE landscape. TE workflows shared a broad core of TE-derived regions but recovered complementary sets of age- and sex-associated TE loci, evaluated with a sex-balanced killifish multi-tissue aging cohort. Together, the combinatorial use of adaptable programs supports robust, locus-aware detection of complex repeats across genome assemblies.

genomics↗

Early origin of sugar sensing in jawed vertebrates

Sweet taste guides animals to consume carbohydrate-rich foods, and many different vertebrate groups, from fish to mammals, rely on sugar-rich fruits or nectar produced by flowering plants (angiosperms). Although the genes encoding T1R2-T1R3, the receptor pair that mammals use to sense sugars, exist in the genomes of many vertebrates, their functions are unclear--and whether sugar sensing arose once early in vertebrate evolution or independently in different lineages after angiosperms evolved is currently unknown. Here, we combined ancestral reconstruction and receptor functional profiling to examine the evolutionary history of T1R taste receptors--including recently-described non-canonical receptors--across all major vertebrate clades. Our results pinpoint the origin of sugar sensing to before the emergence of angiosperms and uncover a myriad of alternative T1R-based sugar-sensing mechanisms, suggesting multiple independent T1R trajectories and revealing uncharted sensory diversity across vertebrates.

evolutionary biology↗

Location dependence of protein intrinsic disorder in Drosophila melanogaster

The relevance of protein structural flexibility in function remains contested, but experimental and computational evidence continues to accumulate. Many efforts to address this investigate intrinsic disorder, which commonly refers to peptide segments or entire protein sequences that presumably lack structure and exhibit high flexibility/conformational heterogeneity under physiological conditions. These efforts face challenges such as conflicting computational predictions and ambiguous relationships among intrinsic disorder locations and other protein properties. We address these challenges at a genome-wide scale in Drosophila melanogaster using residue-level predictions for various protein properties. We employ single and consensus approaches to quantify the prevalence of intrinsic disorder and attempt to infer function by testing for differences along protein sequences. Intrinsic disorder is likely more common at terminals than internal regions, and amino acid frequencies can vary substantially between regions in a manner that plausibly reflects functions of intrinsic disorder, rather than only proteome-wide effects. Tertiary structure potentially underlies the prevalence of intrinsic disorder along protein sequences; this prevalence varies more in a putatively solvent-exposed context than a solvent-buried one. Protein-binding appears to be a main function of intrinsic disorder, and we find support consistent with the notion that structural flexibility fosters binding plasticity, and show that location and protein length are factors in this relationship. Nucleic acid-binding and linker are ostensibly less common disorder functions than protein-binding, but nucleic acid-binding seems more localized at terminals. Residue-level estimates of selection pressure indicate that disordered regions generally evolve under weaker sequence constraints than structured regions, except at the N-terminal region. Biases in disorder prediction are a considerable factor in many of the observations, but unlikely a full explanation. The findings strengthen support for functional relevance of flexibility, offer insight into protein architecture and function, and lend impetus for experimental inquiry.

bioinformatics↗

Shark sexing from forensic, archival, and developmental samples using sex-linked DNA markers

The effective management of threatened shark populations relies on accurate demographic data, particularly operational sex ratios. While sex identification in intact shark bodies is straightforward through the presence of external male organs, namely claspers, it remains impossible for processed fins in the illegal wildlife trade, early-stage embryos in breeding programs, or archived tissue fragments and blood samples where morphological traits are lost. Here, we present a robust molecular sexing framework leveraging recently identified sequences from shark sex chromosomes, consistently organized in the XY system, to our current knowledge. Our approach consists of two distinct methodologies tailored to the the current identification status of sex chromosome sequences in the target species. For the whale shark Rhincodon typus and the brownbanded bamboo shark Chiloscyllium punctatum, we employed end-point PCR assays targeting male-specific Y-linked markers. For the cloudy catshark Scyliorhinus torazame, we developed a quantitative PCR (qPCR) assay targeting differential X chromosome dosage. In this dosage-based system, females (XX) are distinguished by an amplification profile approximately one cycle earlier than males (XY). By integrating X-linked dosage quantification, our framework provides a critical internal control that significantly enhances reliability, allowing researchers to distinguish true females from PCR failures. This toolkit offers a versatile solution for diverse applications, ranging from the study of sex determination mechanisms in pre-phenotypic embryos to the reconstruction of sex ratios from space-constrained tissue archives and global wildlife forensics, thereby contributing to the comprehensive conservation of shark biodiversity.

ecology↗

Inhibition of the gut ceramidase Asah2 decelerates the vertebrate ageing rate

The pace of ageing varies markedly among vertebrate species and individuals. However, the mechanisms underlying these differences in ageing rates remain unclear. Here, we show that the activity of the gut ceramidase Asah2 determines species- and strain-specific rates of vertebrate ageing. Comparative genomic analyses of vertebrate species and strains with different ageing rates reveal an association between low Asah2 activity and increased lifespan. Using the ultra-short-lived killifish Nothobranchius furzeri as a model, we demonstrate that knockout of Asah2 (asah2 KO) extends lifespan and attenuates systemic ageing phenotypes, including declines in locomotor activity, abnormal protein accumulation in the brain, and accumulation of senescent cells in the liver. asah2 KO elevates levels of ceramide species with long-chain fatty acids in the intestine, and supplementation with these ceramide species suppresses ageing phenotypes and extends lifespan in wild-type fish. asah2 KO and ceramide supplementation alter gut microbiota composition, and asah2 KO-derived microbiota transplantation attenuates ageing phenotypes, suggesting that reduced Asah2 activity prevents ageing through intestinal ceramide-mediated modulation of the microbiota. Given the evolutionary conservation of the Asah2 gene and its age-dependent upregulation in fish and humans, Asah2 and ceramides may act as ageing accelerators and decelerators, respectively, across animal species.

developmental biology↗

MEIOSIN retains an intrinsic STRA8-independent activity that drives meiotic entry across vertebrate evolution

Meiotic entry in vertebrates has been viewed as a transcriptional switch driven by the MEIOSIN-STRA8 axis, but whether this represents the ancestral regulatory logic of meiosis has remained unclear. Here we combine comparative genomics with genetic and single-cell analyses in zebrafish and mice to show that MEIOSIN retains an intrinsic STRA8-independent activity. We identify Meiosin orthologs in zebrafish and hagfish, vertebrate lineages that lack Stra8, and show that these proteins retain the HMG domain but have lost the bHLH domain required for the canonical MEIOSIN-STRA8 interaction. In zebrafish, meiosin is transiently induced at meiotic entry in both sexes, yet its loss selectively disrupts the female germ line, causing failure of oogenesis and female-to-male sex reversal. In mice, MEIOSIN lacking the bHLH domain still initiates key features of meiotic entry and partially activates meiotic target genes, although it fails to support full meiotic progression. Together, these findings identify HMG-containing MEIOSIN as a conserved core regulator of meiotic entry and position STRA8 as a later-acting module that enhances the efficiency and robustness of meiotic gene activation.

developmental biology↗

Embryonic and larval development of the Pacific saury Cololabis saira: Distinctive characteristics of a rapidly growing beloniform fish

BackgroundPacific saury Cololabis saira is one of the important food resources drawing attention for its recent rapid decline of catch. Their life cycle and embryonic development have been largely unknown. It is important to clarify how the habitat and reproduction of this species have been affected by the global changes of aquatic environment. ResultsWe obtained fertilized eggs of C. saira, by spontaneous spawning and artificial fertilization, and observed the embryonic development up to larval stages. Embryonic stages are documented with major periods of developmental events; cleavage, gastrulation (epiboly) and somitogenesis and organogenesis. Remarkably, segmentation of somites starts in the middle of epiboly, unlike other well-documented teleost species such as zebrafish and medaka. Morphological changes in larval stage up to feeding juvenile is also described. Growth speed of larval Pacific saury is dramatically rapid, in comparison to closely related beloniform fish such as medaka. ConclusionsIn comparison to medaka, early embryogenesis of saury proceeds slowly, although being followed by early onset of somitogenesis. This might be partly responsible for the rapid growth into adult (larger than 20 cm in body length) in only half a year. Further studies on embryonic development will uncover the molecular mechanisms underlying the characteristics of Pacific saury as an excellent source of nutrition and as an indicator of major environmental changes such as global warming.

developmental biology↗

A Deep Dive into the Globin Superfamily of Sharks, Skates, and Rays: Contrasting patterns of gene loss and retention relative to bony vertebrates

The globin gene superfamily encodes oxygen-binding proteins that are present in all domains of life. Hemoglobins and myoglobins of jawed vertebrates are among the most well-studied proteins in the context of structure-function relationships and evolution after gene duplication. However, these studies have primarily focused on bony vertebrates, and research on globin gene evolution in cartilaginous fish has been limited by a lack of genomic resources. In this study, we leverage newly available cartilaginous fish genomes to investigate globin gene family evolution across skates, rays, sharks, and sawfish. We found that, when present, most globin genes are in a single copy, with androglobin, globin-Y, and myoglobin present in all cartilaginous fish, while the two globin-X genes were differentially retained between the Holocephali and sharks, skates, and rays. Neuroglobin appears to have been lost at the common ancestor of all cartilaginous fish. The - and {beta}-globin gene subfamilies underwent independent expansions in different lineages of cartilaginous fish. Most cartilaginous fish globins have conserved synteny with other jawed vertebrates except myoglobin. Additionally, NPRL3, which directly flanks the hemoglobin clusters of other jawed and jawless vertebrates and regulates hemoglobin gene expression, is on a separate chromosome from the hemoglobin clusters of cartilaginous fish. When we examined globin gene expression patterns across cartilaginous fish tissues and developmental stages, we found that most globins are expressed as expected compared to other jawed vertebrates. However, hemoglobin paralogs are more widely expressed in embryonic tissues compared to later-stage tissues in cases where many copies exist. Our results reveal similar and contrasting patterns of globin gene evolution between cartilaginous and bony vertebrates and shed light on the early stages of globin gene evolution in gnathostomes. Significance StatementThe evolution of the globin gene family in jawed vertebrates is of significant interest; however, most studies have been limited to bony vertebrates. With the influx of new publicly available cartilaginous fish genomes, we conducted the most comprehensive analysis of globin gene evolution in cartilaginous fish to date using phylogenetic, structural, and transcriptomic analyses. Our results shed light on similar and contrasting patterns of globin-gene evolution between cartilaginous and bony vertebrates and provide insight into the early evolution of globins in jawed vertebrates. Key-words: Globins, Cartilaginous Fish, Chondrichthyes, Gene Family Evolution

evolutionary biology↗

Improved genome assembly of whale shark, the world's biggest fish: revealing "chromocline" in intragenomic heterogeneity

High-quality chromosome-level assemblies are essential for understanding genome evolution but remain difficult to obtain for large and complex genomes. Here we present a near gap-free genome assembly of the whale shark (Rhincodon typus) generated with long-read sequencing and Hi-C scaffolding, markedly improving contiguity and completeness. In particular, the X chromosome was extended to nearly twice its previous length, and putative pseudoautosomal regions were identified. Moreover, we report the first Y-linked scaffolds for this species. Comparative analyses with the zebra shark revealed exceptionally low substitution rates across the genome. We further detected a negative correlation between chromosome length and synonymous substitution rate (dS), explained by a positional gradient, designated as "chromocline", in which substitution rates gradually decrease from chromosomal ends toward central regions. Notably, the X chromosome exhibited low dS compared with autosomes of similar size, consistent with male-driven evolution. Our results highlight positional and sex-chromosome effects as key determinants of molecular evolutionary rates. The improved assembly will enable broad application to population-genetic and conservation genomic analyses in the whale shark.

evolutionary biology↗

ATAC-seq of Low-Input and Cryopreserved Primordial Germ Cells Reveals Functional Enhancers

Cell fate specification during development is orchestrated by dynamic changes in gene expression, driven by the accessibility of cis-regulatory elements. To profile such functional regulatory elements at the genome-wide level, bulk ATAC-seq is a powerful method. Yet, its application has still been technically limited in rare cell types. Here, we optimized and titrated the ATAC-seq protocol for cultured chicken primordial germ cells (PGCs) and obtained high-quality chromatin accessibility profiles from as few as 200 cells, even after cryopreservation. Integrative analysis of our ATAC-seq with published RNA-seq and tissue ATAC-seq datasets identified over 10,000 PGC-specific accessible chromatin regions (ACRs), many absent from somatic tissues. Reporter assays demonstrated activity of selected enhancers in cultured PGCs, while transcriptome integration revealed preferential association of PGC-specific ACRs with genes highly expressed at embryonic day 2.5. In vivo transplantation confirmed that these enhancers are active in an early-stage-specific manner, but become silenced upon gonadal settlement. In conclusion, this streamlined method provides a practical and cost-effective approach to the epigenomic analysis of scarce cell populations, facilitating future investigations in developmental biology, germline research, and beyond.

developmental biology↗

Differential fates of vertebrate Kazald gene quartet, from ancestral roles in skeletogenesis and regeneration to putative innovations in fish and birds

Salamanders are known for their incredible regenerative abilities, but translating findings to mammals is complicated by unannotated genes and unclear orthology. An example highlighting this difficulty was a discovery in the axolotl (Ambystoma mexicanum) of a regeneration-associated gene that has been identified as either Kazald1 or Kazald2. Since orthology inference of genes across species is crucial to identifying gains and losses of gene functions, and thus if gene usage is likely to be consistent across species, we investigated the evolution of the axolotl genes using an extensive cross-species analysis. Molecular phylogeny inference conclusively identified the regeneration-associated gene as Kazald2, but also revealed an undescribed four-member Kazald gene family in jawed vertebrates. Moreover, synteny comparisons demonstrated that this family originated in the ancestral two-rounds of whole genome duplication. Additionally, we performed vertebrate-wide comparisons of Kazald gene expression profiles, employing available RNA-Seq which we validated in whole tissues of axolotl, zebrafish, and sharks. This uncovered seemingly ancestral connections conserved over jawed vertebrate evolution, such as Kazald1 with skeletogenesis and odontogenesis and Kazald2 with regeneration. It also suggested novel putative roles within specific lineages, including Kazald3 in teleost fish skeletogenesis and Kazald4 within avian brains. Our study thus demonstrates the establishment of a Kazald gene quartet in the jawed vertebrate ancestor, and elucidates the asymmetry of gene fates of its members, including deeply ancestral roles and comparably recent innovations. This provides a comprehensive report of this formerly undescribed gene family, offering a solid foundation for future studies of these genes in diverse species.

evolutionary biology↗

Tracing genome size dynamics in sharks and rays with inclusive sequence analysis by the Squalomix Consortium

Genomes have maintained stable sets of protein-coding genes during evolution, while chromosome organization and genome size vary drastically. Changes in genome size are often attributed to variable amounts of repetitive sequences, including transposable elements. However, it remains poorly understood how such changes were accommodated while maintaining other genomic components. Elasmobranchs, including sharks, rays, and skates, exhibit high among-species variation of genome size and high within-species variation of chromosome length, offering a unique study system to address the question. In this study, we present the first whole genome sequences of the whitebelly skate with remarkably small genome size among elasmobranchs (2.2 Gb), and the red stingray. These chromosome-scale assemblies enabled the assessment of genomic compositions including centromeres and non-coding elements, which revealed notable profiles of tRNA loci and unbiased intragenomic distribution of transposons in elasmobranch genomes. Comparative analyses across these species revealed a shared genomic architecture characterized by correlations of intergenic and intronic sequence lengths with chromosome sizes, with repetitive element accumulation in elongated regions. We analyzed tandemly duplicated genes with high copy number variability. This genome-wide survey revealed the tendency for more frequent tandem gene duplications along with genome size expansion. We document the batoid HoxC cluster in the red stingray genome, which has undergone extensive repetitive element invasion and co-localizes with the HoxB cluster on a sex chromosome. Our study demonstrates an inclusive analysis encompassing both coding and non-coding regions, adaptable to more species in the taxon and a basis for molecular-level understanding on phenotypic diversity of elasmobranchs.

genomics↗

RNA Dynamics Regulate Transcriptional Condensate Vivacity to Drive Gene Coordination

Transcriptional condensates (TCs), enriched with Mediator, orchestrate super-enhancer (SE)-driven gene expression critical for cell identity. However, how their dynamic physical properties shape transcriptional activities remains unclear. Here, we reveal a previously unknown regulatory axis wherein dynamic features of enhancer RNA (eRNA), transcribed but rapidly degraded by the RNA exosome, maintain optimal TC fluidity and stability. Depletion of RNA exosome disrupts TC integrity, altering Mediator and Pol II colocalization, in embryonic stem cells. This TC perturbation attenuates proper transcriptional regulator loading and pause-release regulation, enhances transcriptional noise, and diminishes coordinated eRNA-mRNA expression for SE-associated genes. Multi-omics analyses, live-cell imaging, and computational modeling collectively demonstrate that RNA turnover modulates TC vivacity, facilitating widespread chromatin contacts between SE-containing active A compartments and coordinated transcriptional bursts across SE-associated genes. Our findings establish RNA-dependent condensate dynamics as an essential quality-control mechanism that fine-tunes global transcriptional coordination, maintaining cellular homeostasis and cell identity. HighlightsO_LIRNA exosome is required for the homeostasis of transcriptional condensates (TCs) C_LIO_LIRNA synthesis and decay optimize the fluidity and dynamic properties of TCs C_LIO_LITCs maintain the transcriptional consistency of super-enhancer (SE) genes C_LIO_LITCs support contacts of SE-containing A compartments for coordinated transcription C_LI

molecular biology↗

Empowering medaka fish biology with versatile genomic resources in MedakaBase

Medaka, a group of small, mostly freshwater fishes in the teleost order Beloniformes, includes the rice fish Oryzias latipes, which is a prominent model organism for diverse biological fields. Chromosome-scale genome sequences of the Hd-rR strain of this species were obtained in 2007, and its improved version has facilitated various genome-wide studies. However, despite its widespread utility, omics data for O. latipes are dispersed across various public databases and lack a centralized platform. To address this, the medaka section of the National Bioresource Project (NBRP) of Japan established a genome informatics team in 2022 tasked with providing versatile in silico solutions for bench biologists. This initiative led to the launch of MedakaBase (https://medakabase.nbrp.jp), a web server that enables gene-oriented analysis including exhaustive sequence similarity searches. MedakaBase also provides genome-wide browsing of diverse datasets, including tissue-specific transcriptomes and intraspecific genomic variations, integrated with gene models from different sources. Additionally, the platform offers gene models optimized for single-cell transcriptome analysis, which often requires coverage of the 3' untranslated region (UTR) of transcripts. Currently, MedakaBase provides genome-wide data for seven Oryzias species, including original data for O. mekongensis and O. luzonensis produced by the NBRP team. This article outlines technical details behind the data provided by MedakaBase.

genomics↗

Ancestral origin and structural characteristics of non-syntenic homologous chromosomes in abalones

Structural variation is increasingly recognized as a pivotal contributor to genomic diversity in marine invertebrates, yet its extent and evolutionary significance remain poorly characterized in many species. Haplotype-phased genome assembly is an excellent method for studying such variations by comparing homologous chromosomes. In this study, we constructed a haplotype-phased genome assembly for the western Pacific abalone, Haliotis gigantea, using high-fidelity (HiFi) long-read sequencing and high-resolution chromosome conformation capture (Hi-C) data. The primary and alternative assemblies each comprised 18 long scaffolds (>50 Mb), consistent with the species diploid chromosome number (2n = 36), and contained 96.5% and 96.2% complete single-copy Metazoa Benchmarking Universal Single-Copy Orthologs genes, respectively, indicating high assembly quality. Comparative analysis of the two haplotypes revealed three homologous chromosomes with large-scale non-syntenic regions caused by extensive segmental duplications, with each enriched in distinct gene domains. These non-syntenic chromosomes likely originated in abalone evolution, as they were conserved across both closely and distantly related species. Our findings highlight the evolutionary importance of non-syntenic structural variation in shaping genome architecture and suggest that such variation may play a broader role in functional diversification across abalones.

genomics↗

Sharks and rays have the oldest vertebrate sex chromosome with unique sex determination mechanisms

Sex determination has been investigated across vertebrate lineages to reveal stepwise evolution of sex chromosomes and diversity of responsible molecular mechanisms. However, these studies hardly encompass cartilaginous fishes deeply isolated from the rest of vertebrates, which hinders the comprehensive view of vertebrate sex determination. Here, we produced chromosome-scale genome assemblies of egg-laying shark species and comparatively investigated genome sequences and transcriptome profiles across diverse cartilaginous fishes. Sex chromosome identification, supported by cytogenetic experiments, elucidated the homology of X chromosomes between sharks and rays as well as an extensively degenerating Y chromosome harboring no male-specific genes. These sex chromosomes scarcely included orthologs of previously documented sex determining genes. Transcriptomic analyses combined with histology of embryonic gonads revealed female-biased expressions of X-linked genes including those implicated in TGF-{beta} and IGF signaling pathways, which are attributed to their incomplete dosage compensation. Our findings indicate that sharks and rays share the oldest vertebrate sex chromosomes that originated around 300 million years ago and the dosage-dependent sex determination mechanism comprised of distinct molecules from other vertebrates. This study highlights the antiquity of sex chromosomes and uniqueness of sex determination mechanisms in sharks and rays, which advance our understanding on evolutionary plasticity of vertebrate sex determination.

evolutionary biology↗

The Greenland shark genome: insights into deep-sea ecology and lifespan extremes

The Greenland shark (Somniosus microcephalus) is known for its slow metabolism and deep-sea habitat. It is considered the longest-lived vertebrate on Earth, with an estimated lifespan of 392{+/-}120 years. Despite its remarkable longevity and lifestyle, there have been no genomic studies on this species. Here, we report the first, chromosome-level assembly of the Greenland shark genome, which is 5.9 Gb in size with an N50 length of 233 Mb, and contains 37,125 predicted genes with a completeness score of 86.5%. We found that the copy numbers of three gene families (TNF, TLR, LRRFIP), which are involved in activating the NF-{kappa}B signaling pathway, are significantly increased in the Greenland shark compared to short-lived shark species. In the rhodopsin of this deep-sea dweller, we detected amino acid substitutions that result in spectral tuning for the so-called blue shift, suggesting adaptive evolution to dim-light vision. We also elucidate the dynamics of the effective population size (Ne) of the Greenland shark and its close relative, the Pacific sleeper shark (Somniosus pacificus). These genomic analyses offer new insights into the molecular basis of the exceptional longevity of the Greenland shark and highlight potential genetic mechanisms that could inform future research into longevity.

genomics↗

Silver chimaera genome assembly and identification of the holocephalan sex chromosome sequence

Cartilaginous fishes are divided into holocephalans and elasmobranchs, and comparative studies involving them are expected to elucidate how variable phenotypes and distinctive genomic properties were established in those ancient vertebrate lineages. To date, molecular-level studies on holocephalans have concentrated on the family Callorhinchidae, with a chromosome-scale genome assembly of Callorhinchus milii available. In this study, we focused on the most species-rich holocephalan family Chimaeridae and sequenced the genome of its member, silver chimaera (Chimaera phantasma). We report the first chromosome-scale genome assembly of the Chimaeridae, with high continuity and completeness, which exhibited a large intragenomic variation of chromosome lengths, which is correlated with intron size. This pattern is observed more widely in vertebrates and at least partly accounts for cross-species genome size variation. A male-female comparison identified a silver chimaera genomic scaffold with a double sequence depth for females, which we identify as an X chromosome fragment. This is the first DNA sequence-based evidence of a holocephalan sex chromosome, suggesting a male heterogametic sex determination system. This study, allowing the first chromosome-level comparison among holocephalan genomes, will trigger in-depth understanding of the genomic diversity among vertebrates as well as species population genetic structures based on the genome assembly of high completeness.

genomics↗