ABSTRACT
Coprolites contain various kinds of ancient DNAs derived from gut micro-organisms, viruses, and foods, which can help to determine the gut environment of ancient peoples. Their genomic information should be helpful in elucidating the interaction between hosts and microbes for thousands of years, as well as characterizing the dietary behaviors of ancient people. We performed shotgun metagenomic sequencing on four coprolites excavated from the Torihama shell-mound site in the Japanese archipelago. The coprolites were found in the layers of the Early Jomon period, corresponding stratigraphically to 7000 to 5500 years ago. After shotgun sequencing, we found that a significant number of reads showed homology with known gut microbe, viruses, and food genomes typically found in the feces of modern humans. We detected reads derived from several types of phages and their host bacteria simultaneously, suggesting the coexistence of viruses and their hosts. The food genomes provide biological evidence for the dietary behavior of the Jomon people, consistent with previous archaeological findings. These results indicate that ancient genomic analysis of coprolites is useful for understanding the gut environment and lifestyle of ancient peoples.
Subject(s)
Metagenome , Metagenomics , Humans , Japan , Genomics , ArchaeologyABSTRACT
In various epithelial tissues, the epithelial monolayer acts as a barrier. To fulfill its function, the structural integrity of the epithelium is tightly controlled. When normal epithelial cells detach from the basal substratum and delaminate into the apical lumen, the apically extruded cells undergo apoptosis, which is termed anoikis. In contrast, transformed cells often become resistant to anoikis and able to survive and grow in the apical luminal space, leading to the formation of multilayered structures, which can be observed at the early stage of carcinogenesis. However, the underlying molecular mechanisms still remain elusive. In this study, we first demonstrate that S100A10 and ANXA2 (Annexin A2) accumulate in apically extruded, transformed cells in both various cell culture systems and murine epithelial tissues in vivo. ANXA2 acts upstream of S100A10 accumulation. Knockdown of ANXA2 promotes apoptosis of apically extruded RasV12-transformed cells and suppresses the formation of multilayered epithelia. In addition, the intracellular reactive oxygen species (ROS) are elevated in apically extruded RasV12 cells. Treatment with ROS scavenger Trolox reduces the occurrence of apoptosis of apically extruded ANXA2-knockdown RasV12 cells and restores the formation of multilayered epithelia. Furthermore, ROS-mediated p38MAPK activation is observed in apically delaminated RasV12 cells, and ANXA2 knockdown further enhances the p38MAPK activity. Moreover, the p38MAPK inhibitor promotes the formation of multilayered epithelia of ANXA2-knockdown RasV12 cells. These results indicate that accumulated ANXA2 diminishes the ROS-mediated p38MAPK activation in apically extruded transformed cells, thereby blocking the induction of apoptosis. Hence, ANXA2 can be a potential therapeutic target to prevent multilayered, precancerous lesions.
Subject(s)
Annexin A2 , Animals , Mice , Annexin A2/genetics , Apoptosis , Epithelial Cells , Epithelium , Reactive Oxygen SpeciesABSTRACT
Recent studies have shown that adipose tissue is an immunological organ. While inflammation in energy-storing white adipose tissues has been the focus of intense research, the regulatory mechanisms of inflammation in heat-producing brown adipose tissues remain largely unknown. We previously identified apoptosis signal-regulating kinase 1 (ASK1) as a critical regulator of brown adipocyte maturation; the PKA-ASK1-p38 axis facilitates uncoupling protein 1 (UCP1) induction cell-autonomously. Here, we show that ASK1 suppresses an innate immune pathway and contributes to maintenance of brown adipocytes. We report a novel chemical pull-down method for endogenous kinases using analog sensitive kinase allele (ASKA) technology and identify an ASK1 interactor in brown adipocytes, receptor-interacting serine/threonine-protein kinase 2 (RIPK2). ASK1 disrupts the RIPK2 signaling complex and inhibits the NOD-RIPK2 pathway to downregulate the production of inflammatory cytokines. As a potential biological significance, an in vitro model for intercellular regulation suggests that ASK1 facilitates the expression of UCP1 through the suppression of inflammatory cytokine production. In parallel to our previous report on the PKA-ASK1-p38 axis, our work raises the possibility of an auxiliary role of ASK1 in brown adipocyte maintenance through neutralizing the thermogenesis-suppressive effect of the NOD-RIPK2 pathway.
Subject(s)
Adipocytes, Brown/metabolism , MAP Kinase Kinase Kinase 5/pharmacology , Nod Signaling Adaptor Proteins/drug effects , Receptor-Interacting Protein Serine-Threonine Kinase 2/drug effects , Adipocytes, Brown/drug effects , Adipocytes, White/metabolism , Animals , Cytokines/analysis , HEK293 Cells , Humans , Inflammation/drug therapy , Mice , Receptor-Interacting Protein Serine-Threonine Kinase 2/metabolism , Signal Transduction/drug effects , Uncoupling Protein 1/drug effectsABSTRACT
BACKGROUND: Ring finger protein 213 (RNF213) is a susceptibility gene of moyamoya disease (MMD). A previous case-control study and a family analysis demonstrated a strong association of the East Asian-specific variant, R4810K (rs112735431), with MMD. Our aim is to uncover evolutionary history of R4810K in East Asian populations. METHODS: The RNF213 locus of 24 MMD patients in Japan were sequenced using targeted-capture sequencing. Based on the sequence data, we conducted population genetic analysis and estimated the age of R4810K using coalescent simulation. RESULTS: The diversity of the RNF213 gene was higher in Africans than non-Africans, which can be explained by bottleneck effect of the out-of-Africa migration. Coalescent simulation showed that the risk variant was born in East Asia 14,500-5100 years ago and came to the Japanese archipelago afterward, probably in the period when the known migration based on archaeological evidences occurred. CONCLUSIONS: Although clinical data show that the symptoms varies, all sequences harboring the risk allele are almost identical with a small number of exceptions, suggesting the MMD phenotypes are unaffected by the variants of this gene and rather would be more affected by environmental factors.
Subject(s)
Adenosine Triphosphatases/genetics , Moyamoya Disease/genetics , Ubiquitin-Protein Ligases/genetics , Alleles , Evolution, Molecular , Gene Frequency , Genetics, Population , Haplotypes , Humans , Japan , Linkage DisequilibriumABSTRACT
Ferroptosis has recently attracted much interest because of its relevance to human diseases such as cancer and ischemia-reperfusion injury. We have reported that prolonged severe cold stress induces lipid peroxidation-dependent ferroptosis, but the upstream mechanism remains unknown. Here, using genome-wide CRISPR screening, we found that a mitochondrial Ca2+ uptake regulator, mitochondrial calcium uptake 1 (MICU1), is required for generating lipid peroxide and subsequent ferroptosis under cold stress. Furthermore, the gatekeeping activity of MICU1 through mitochondrial calcium uniporter (MCU) is suggested to be indispensable for cold stress-induced ferroptosis. MICU1 is required for mitochondrial Ca2+ increase, hyperpolarization of the mitochondrial membrane potential (MMP), and subsequent lipid peroxidation under cold stress. Collectively, these findings suggest that the MICU1-dependent mitochondrial Ca2+ homeostasis-MMP hyperpolarization axis is involved in cold stress-induced lipid peroxidation and ferroptosis.
Subject(s)
Cation Transport Proteins , Ferroptosis , Calcium/metabolism , Calcium-Binding Proteins/metabolism , Cation Transport Proteins/metabolism , Cold-Shock Response , Humans , Membrane Potential, Mitochondrial , Mitochondrial Membrane Transport Proteins/genetics , Mitochondrial Membrane Transport Proteins/metabolismABSTRACT
Cell competition is a social cellular phenomenon in which unfit cells are selectively eliminated to maintain tissue homeostasis.1-3 Recent studies have revealed that mechanical forces induce competitive cell-cell interactions in Drosophila.4-6 This mechanical cell competition has also been reported to play an important role in mammalian cells, using Madin-Darby canine kidney (MDCK) cells depleted of a polarity regulator Scribble in a tetracycline-inducible manner (scribKD cells).7scribKD cells are hypersensitive to crowding due to the lower homeostatic density than wild-type (WT) cells,7,8 and in the context of cell competition, scribKD cells are compacted and eliminated by WT cells.7-10 Although p38 and p53 are involved in this process,7,10 the molecular mechanism by which WT cells recognize and mechanically eliminate scribKD cells remains unclear. Here, we report that scribKD cells secrete fibroblast growth factor 21 (FGF21) to drive cell competition. Knockdown of FGF21 in scribKD cells or loss of FGFR1 in WT cells suppresses cell competition, suggesting that WT cells recognize scribKD cells through FGF21. FGF21-containing culture medium of scribKD cells activates cell motility. Moreover, FGF21 promotes the compression and elimination of scribKD cells by attracting surrounding WT cells. We also demonstrate that activation of the apoptosis signal-regulating kinase 1 (ASK1)-p38 pathway in scribKD cells induces FGF21 to drive cell competition. Our findings reveal a mechanism whereby WT cells mechanically eliminate scribKD cells and propose a new function for FGF21 in cell-cell communication.
Subject(s)
Cell Competition , Drosophila Proteins , Animals , Dogs , Drosophila , Drosophila Proteins/genetics , Fibroblast Growth Factors , Madin Darby Canine Kidney CellsABSTRACT
To analyze a specific genome region using next-generation sequencing technologies, the enrichment of DNA libraries with targeted capture methods has been standardized. For enrichment of mitochondrial genome, a previous study developed an original targeted capture method that use baits constructed from long-range polymerase chain reaction (PCR) amplicons, common laboratory reagents, and equipment. In this study, a new targeted capture method is presented, that of bacterial artificial chromosome (BAC) double capture (BDC), modifying the previous method, but using BAC libraries as baits for sequencing a relatively large gene. We applied the BDC approach for the 214 kb autosomal region, ring finger protein 213, which is the susceptibility gene of moyamoya disease (MMD). To evaluate the reliability of BDC, cost and data quality were compared with those of a commercial kit. While the ratio of duplicate reads was higher, the cost was less than that of the commercial kit. The data quality was sufficiently the same as that of the kit. Thus, BDC can be an easy, low-cost, and useful method for analyzing individual genome regions with substantial length.
Subject(s)
Chromosomes, Artificial, Bacterial/genetics , Gene Library , High-Throughput Nucleotide Sequencing/methods , Sequence Analysis, DNA/methods , Adenosine Triphosphatases/genetics , Genetic Predisposition to Disease , Humans , Moyamoya Disease/genetics , Ubiquitin-Protein Ligases/geneticsABSTRACT
Parasite-induced behavioral changes in their hosts favor to complete the lifecycle of parasites. Schistosome infection is also known to cause physiological changes in infected freshwater snail intermediate hosts. Here, we report, a novel phenomenon in which Schistosoma mansoni, a highly debilitating worm affecting millions of people worldwide, alters the phototropic behavior of Biomphalaria glabrata, the vector snail. S. mansoni-infection enhanced positive phototropism of vector snails and infected snails spent significantly more time in light. Possibly, these behavioral changes help the parasite to be released efficiently from the infected intermediate hosts, and to infect mammalian hosts.
Subject(s)
Behavior, Animal , Biomphalaria/physiology , Biomphalaria/parasitology , Host-Parasite Interactions , Phototropism , Schistosomiasis mansoni/veterinary , Animals , Disease Vectors , Light , Schistosoma mansoni , Water/parasitologyABSTRACT
Cilia and flagella are hair-like organelles that project from the cell surface and play important roles in motility and sensory perception. Motility defects in cilia and flagella lead to primary ciliary dyskinesia (PCD), a rare human disease. Recently zinc finger MYND-type containing 10 (ZMYND10) was identified in humans as a PCD-associated gene. In this study, we use medaka fish as a model to characterize the precise functions of zmynd10. In medaka, zmynd10 is exclusively expressed in cells with motile cilia. Embryos with zmynd10 Morpholino knockdown exhibited a left-right (LR) defect associated with loss of motility in Kupffer's vesicle (KV) cilia. This immotility was caused by loss of the outer dynein arms, which is a characteristic ultrastructural phenotype in PCD. In addition, KV cilia in zmynd10 knockdown embryos had a swollen and wavy morphology. Together, these results suggest that zmynd10 is a multi-functional protein that has independent roles in axonemal localization of dynein arms and in formation and/or maintenance of cilia. The C-terminal region of zmynd10 has a MYND-type zinc finger domain (zf-MYND) that is important for its function. Our rescue experiment showed that the zmynd10-ΔC truncated protein, which lacks zf-MYND, was still partially functional, suggesting that zmynd10 has another functional domain besides zf-MYND. To analyze the later stages of development, we generated a zmynd10 knockout mutant using transcription activator-like effector nuclease (TALEN) technology. Adult mutants exhibited sperm dysmotility, scoliosis and progressive polycystic kidney.
Subject(s)
Axoneme/metabolism , Cilia/metabolism , Dyneins/metabolism , Oryzias/metabolism , Polycystic Kidney Diseases/metabolism , Scoliosis/metabolism , Amino Acid Sequence , Animals , Axoneme/drug effects , Base Sequence , Body Patterning/drug effects , Body Patterning/genetics , Cilia/drug effects , Embryo, Nonmammalian/drug effects , Embryo, Nonmammalian/metabolism , Epistasis, Genetic/drug effects , Gene Expression Regulation, Developmental/drug effects , Male , Morpholinos/pharmacology , Movement , Oryzias/embryology , Oryzias/genetics , Phenotype , Polycystic Kidney Diseases/pathology , RNA, Messenger/genetics , RNA, Messenger/metabolism , Scoliosis/pathology , Spermatozoa/metabolism , Tumor Suppressor Proteins/chemistry , Tumor Suppressor Proteins/genetics , Tumor Suppressor Proteins/metabolism , Zinc FingersABSTRACT
OBJECTIVES: A cline of frequencies of the derived allele of the ALDH2 gene, which causes a deficiency of an enzyme and "facial flushing" in humans who drink alcohol, has been known among the people of the Japanese archipelago. This cline is conventionally explained by admixture with immigrants from the Asian continent occurring during the Yayoi period. Previous studies lack sufficient data from the peripheral regions of the indigenous Jomon people, and those data the ADH1B gene that is involved in the Class I ADH gene cluster and contains another variant leading to a functional change. METHODS: We focused on the southwestern-most people from the Ryukyu Islands (n = 218) and those from northern Kyushu (n = 21) where the Yayoi immigrants likely arrived. We investigated both the Class I ADH and ALDH2 loci, as well as neutral genetic markers. RESULTS: In the Ryukyu Islands, the frequencies of the ancestral alleles in both loci were always higher than those in mainland Japan, while the frequencies of ADH1B were less than those of the derived allele. A haplotype block was not observed in ALDH2 but was in Class I ADH. DISCUSSION: Our data suggest that the derived allele of ALDH2 came with the Yayoi immigrants from the Asian continent to the Japanese archipelago. However, the derived allele of ADH1B is unlikely to be related to the Yayoi migration. Therefore, we postulate that the expansion of the derived allele of ADHIB in East Asia could be traced back to the last glacial period.
Subject(s)
Alcohol Dehydrogenase/genetics , Aldehyde Dehydrogenase, Mitochondrial/genetics , Gene Frequency , Polymorphism, Genetic , Asia, Eastern , Female , Humans , Islands , Japan , MaleABSTRACT
Radiotherapy is widely used in cancer treatment. In addition to inducing effects in the irradiated area, irradiation may induce effects on tissues close to and distant from the irradiated area. Japanese medaka, Oryzias latipes, is a small teleost fish and a model organism for evaluating the environmental effects of radiation. In this study, we applied low-energy carbon-ion (26.7 MeV/u) irradiation to adult medaka to a depth of approximately 2.2 mm from the body surface using an irradiation system at the National Institutes for Quantum and Radiological Science and Technology. We histologically evaluated the systemic alterations induced by irradiation using serial sections of the whole body, and conducted a heart rate analysis. Tissues from the irradiated side showed signs of serious injury that corresponded with the radiation dose. A 3D reconstruction analysis of the kidney sections showed reductions in the kidney volume and blood cell mass along the irradiated area, reflecting the precise localization of the injuries caused by carbon-beam irradiation. Capillary aneurysms were observed in the gill in both ventrally and dorsally irradiated fish, suggesting systemic irradiation effects. The present study provides an in vivo model for further investigation of the effects of irradiation beyond the locally irradiated area.
Subject(s)
Heavy Ion Radiotherapy/adverse effects , Kidney/pathology , Myocardium/pathology , Oryzias/metabolism , Radiation Injuries, Experimental/pathology , Animals , Kidney/metabolism , Myocardium/metabolism , Radiation Injuries, Experimental/metabolismABSTRACT
Fertilization begins with species-restricted interaction of sperm and the egg-coating envelope, which includes a three-dimensional meshwork of filaments composed of glycoproteins (called ZP proteins). Growing evidence has unveiled the molecular nature of ZP proteins; however, the structural property conferring fertilization competence to the egg-coating envelope remains unknown. Here, we show the molecular mechanism that mediates direct interaction between dicalcin, a novel fertilization-suppressive ZP protein-associated protein, and gp41, a Xenopus laevis ortholog of mammalian ZP3, and subsequently demonstrate the structural basis of the envelope for fertilization competence. The interactive regions between dicalcin and gp41 comprised five and nine amino acid residues within dicalcin and twenty-three within gp41 [corrected]. Synthetic peptides corresponding to these regions dramatically affected fertilization: treatment with dicalcin- or gp41-derived peptides decreased or increased fertilization rates, respectively. Prior application of these peptides caused distinct alterations in the in vivo lectin-staining pattern of the envelope as well. Transmission electron microscopy analysis revealed that the dicalcin-derived peptide induced the formation of a well-organized meshwork, whereas the gp41-derived peptide caused the formation of a significantly disorganized meshwork. These findings indicated that the fertilization competence of the egg-coating envelope is crucially regulated by the direct interaction between dicalcin and gp41.
Subject(s)
Fertilization , Glycoproteins/physiology , S100 Proteins/physiology , Xenopus Proteins/physiology , Xenopus laevis/physiology , Amino Acid Sequence , Animals , Cells, Cultured , Glycoproteins/chemistry , Microscopy, Electron, Transmission , Molecular Sequence Data , Ovum/physiology , Ovum/ultrastructure , Peptide Fragments/chemistry , Protein Binding , Protein Interaction Domains and Motifs , S100 Proteins/chemistry , Sperm-Ovum Interactions , Xenopus Proteins/chemistryABSTRACT
Fertilization comprises oligosaccharide-mediated sperm-egg interactions, including sperm binding to an extracellular egg envelope, sperm penetration through the envelope, and fusion with an egg plasma membrane. We show that Xenopus dicalcin, an S100-like Ca(2+)-binding protein, present in the extracellular egg envelope (vitelline envelope (VE)), is a suppressive mediator of sperm-egg interaction. Preincubation with specific antibody greatly increased the efficiency of in vitro fertilization, whereas prior application of exogenous dicalcin substantially inhibited fertilization as well as sperm binding to an egg and in vitro sperm penetration through the VE protein layer. Dicalcin showed binding to protein cores of gp41 and gp37, constituents of VE, in a Ca(2+)-dependent manner and increased in vivo reactivity of VE with a lectin, Ricinus communis agglutinin I, which was accounted for by increased binding ability of gp41 to the lectin and greater exposure of gp41 to an external environment. Our findings strongly suggest that dicalcin regulates the distribution of oligosaccharides within the VE through its binding to the protein core of gp41, probably by modulating configuration of oligosaccharides on gp41 and the three-dimensional structure of VE framework, and thereby plays a pivotal role in sperm-egg interactions during fertilization.
Subject(s)
Fertilization , Glycoproteins/metabolism , Ovum/metabolism , S100 Proteins/physiology , Xenopus Proteins/physiology , Xenopus laevis/physiology , Animals , Calcium/metabolism , Chromatography, Affinity , Escherichia coli/genetics , Glycosylation , Immunohistochemistry , Recombinant Proteins/metabolism , S100 Proteins/genetics , S100 Proteins/metabolism , Spectrometry, Fluorescence , Sperm-Ovum Interactions , Xenopus Proteins/genetics , Xenopus Proteins/metabolismABSTRACT
The sex-determining region Y (SRY) gene and its related Sox genes encode transcriptional regulatory factors. In this study, we isolated and sequenced a novel Sox cDNA from African clawed frog (Xenopus laevis). The Sox gene was named xSox33. xSox33 was revealed to encode 244 amino acids. Reverse transcription-polymerase chain reaction (RT-PCR) showed that xSox33 was expressed at very low levels in some frog tissues including lung, ovary, skeletal muscle, testis, brain and heart. Its embryonic expression was also studied by RT-PCR. After the mid-blastula transition, the zygotic expression was initiated during gastrulation and the level was elevated as the embryogenesis proceeded. Electrophoretic mobility shift assay (EMSA) indicated that a recombinant xSox33 polypeptide was capable of binding to the nucleotide sequence AACAAT.
Subject(s)
Genes, sry , High Mobility Group Proteins/genetics , Transcription Factors/genetics , Xenopus Proteins/genetics , Xenopus laevis/genetics , Amino Acid Sequence , Animals , Base Sequence , DNA, Complementary/chemistry , DNA, Complementary/isolation & purification , DNA-Binding Proteins/genetics , Electrophoretic Mobility Shift Assay , Molecular Sequence Data , Reverse Transcriptase Polymerase Chain Reaction , SOX Transcription Factors , Sequence Alignment , Xenopus laevis/embryologyABSTRACT
Paxillin has been recognized as a focal adhesion adapter protein that participates in the integrin-mediated signaling. An earlier study [Ogawa et al. Biochim. Biophys. Acta 1519 (2001) 235] found that frog paxillin was expressed in the kidney epithelial cell line A6 and localized in the nucleus. Here, in this study, we have found that the expression of frog paxillin is up-regulated in the S phase of cell cycle. The protein became phosphorylated on tyrosine when the cells were grown on vitronectin; the tyrosine phosphorylation was not detectable when the cells were cultured on fibronectin, laminin or poly-D-lysine. On the other hand, MAP kinase was revealed to phosphorylate frog paxillin on serine. Both phosphorylation events, namely on tyrosine and serine, were essential for the nuclear translocation of this protein. Our results suggest that the integrin-mediated signaling pathway and the MAP kinase pathway meet at paxillin.
Subject(s)
Active Transport, Cell Nucleus/physiology , Cytoskeletal Proteins/metabolism , Phosphoproteins/metabolism , Xenopus laevis/physiology , Animals , Antineoplastic Agents/metabolism , Apigenin , Cell Cycle/physiology , Cell Fractionation , Cell Line , Cytoskeletal Proteins/genetics , Fibronectins/metabolism , Flavonoids/metabolism , Integrins/metabolism , Laminin/metabolism , Mitogen-Activated Protein Kinases/metabolism , Models, Biological , Paxillin , Phosphoproteins/genetics , Phosphorylation , Serine/metabolism , Tyrosine/metabolism , Vitronectin/metabolism , Xenopus ProteinsABSTRACT
Human ENX-2 is a homologue of Drosophila Enhancer of zeste, which is a member of Polycomb-group proteins regulating the expression of homeotic genes as chromatin-associated proteins. In this study, we demonstrate that ENX-2 plays an important role as a signaling molecule involved in T cell receptor-mediated signaling pathway. In immunoprecipitation experiments, ENX-2 and zeta associated protein-70 (ZAP-70) were co-precipitated from T cell lysate. When probed with an anti-phospho-tyrosine antibody, ENX-2 was found to be phosphorylated on tyrosine. On the other hand, ENX-2 was not phosphorylated on tyrosine in the mutant Jurkat cell, J.Cam1.6 lacking the activity of lymphocyte protein tyrosine kinase p56(lck). The interaction between ENX-2 and ZAP-70 was abolished in the mutant cell. Furthermore, in-vitro kinase assay using purified p56(lck) demonstrated that ENX-2 became tyrosine phosphorylated by this kinase. These findings show that the phosphorylation of ENX-2 is responsible for the interaction between ENX-2 and ZAP-70.
Subject(s)
Protein-Tyrosine Kinases/metabolism , Repressor Proteins/metabolism , Signal Transduction/immunology , Blotting, Western , Electrophoresis, Polyacrylamide Gel , Humans , Jurkat Cells/physiology , Lymphocyte Specific Protein Tyrosine Kinase p56(lck)/genetics , Macromolecular Substances , Mutation , Phosphorylation , Polycomb-Group Proteins , Precipitin Tests , Receptors, Antigen, T-Cell/chemistry , Receptors, Antigen, T-Cell/physiology , T-Lymphocytes/physiology , Tyrosine/metabolism , ZAP-70 Protein-Tyrosine KinaseABSTRACT
Sox is a large family of genes related to the sex-determining region Y gene (designated as the SRY gene). Sox genes encoding DNA-binding transcriptional factors are found in many animals and are involved in developmental events. In this study, we newly isolated and sequenced novel Sox cDNAs from African clawed frog (Xenopus laevis). Five clones isolated here were classified into four distinct Sox genes designated as xSox17alpha1, xSox17alpha2, xSox18alpha and xSox18beta. All four belong to a subtype of SOX family, type F. The cDNA xSox17alpha1 contains essentially the same nucleotide sequence as that identified as Sox17alpha in a previous work (Cell 91 (1997) 397), whereas xSox17alpha2 is a distinct gene with high homology to xSox17alpha1. The clones, xSox18alpha and xSox18beta, are highly homologous to each other over the entire nucleotide sequences. The xSox18alpha and xSox18beta genes encode 363 and 361 amino acids, respectively. Genomic Southern hybridization analysis showed the existence of two copies of the xSox18. Northern analysis indicated that the xSox18 gene was expressed in the spleen and kidney and the size of the transcript was estimated to be 2.4 knt. Electrophoretic mobility shift assays indicated that recombinant xSox18 polypeptide was capable of binding to the HMG consensus nucleotide sequence, AACAAT.