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Kazuho Ikeo

Publications and source records attributed to Kazuho Ikeo.

At least 19 recordsLinked to original sources

Direct link between convergent evolution at sequence level and phenotypic level of septal pore cap in Agaricomycotina.

Several homologous morphological characters, despite sharing apparently similar features, are known to have independently evolved in different lineages multiple times. However, the genetic backgrounds of such morphological convergences remain poorly understood. To detect any correlated amino acid substitutions potentially responsible for morphological convergence at the phenotypic level, we focused on the morphology of the septal pore cap (SPC), a structure involved in mycelia's complex multicellularity in fungi. SPCs are classified into 3 morphological types: perforate, imperforate, and vesiculate. To understand the evolutionary events that occurred at the sequence level during the morphological convergence of perforate SPCs in Agaricomycotina, we examined sequence differences among species with different SPC types by comparative genomic analysis using a single-copy gene dataset from 12 Agaricomycotina genomes with morphological literature of SPC. Our analysis revealed that sequences of 8 genes, including an SPC-related gene spc33, were clustered based on SPC morphology rather than species relationship. Additionally, same amino acid substitutions independently occurred in both lineages in which species with perforate SPCs emerged. These findings suggest that specific amino acid substitutions in spc33 were critical for the emergence of perforate SPCs in multiple lineages. Further, our gene search for spc33 across organisms suggests that spc33 evolved shortly before the emergence of imperforate SPC. This study represents the first step toward elucidating the genetic basis of the morphological evolution of SPC. It contributes to both clarifying the genetic basis underlying morphological convergence and advances the study of fungal evolutionary morphology.

Evolution, Molecular↗

Curated genome annotation of Oryza sativa ssp. japonica and comparative genome analysis with Arabidopsis thaliana.

We present here the annotation of the complete genome of rice Oryza sativa L. ssp. japonica cultivar Nipponbare. All functional annotations for proteins and non-protein-coding RNA (npRNA) candidates were manually curated. Functions were identified or inferred in 19,969 (70%) of the proteins, and 131 possible npRNAs (including 58 antisense transcripts) were found. Almost 5000 annotated protein-coding genes were found to be disrupted in insertional mutant lines, which will accelerate future experimental validation of the annotations. The rice loci were determined by using cDNA sequences obtained from rice and other representative cereals. Our conservative estimate based on these loci and an extrapolation suggested that the gene number of rice is approximately 32,000, which is smaller than previous estimates. We conducted comparative analyses between rice and Arabidopsis thaliana and found that both genomes possessed several lineage-specific genes, which might account for the observed differences between these species, while they had similar sets of predicted functional domains among the protein sequences. A system to control translational efficiency seems to be conserved across large evolutionary distances. Moreover, the evolutionary process of protein-coding genes was examined. Our results suggest that natural selection may have played a role for duplicated genes in both species, so that duplication was suppressed or favored in a manner that depended on the function of a gene.

Arabidopsis↗

Degeneration after sexual differentiation in hydra and its relevance to the evolution of aging.

Aging occurs in most multicellular animals, yet some primitive animals do not show any sign of aging. This raises the following question: How have metazoans acquired the trait of aging in the course of evolution? Comparative studies of various species have provided a clue to this question by showing that sexually reproducing organisms predominantly undergo aging. The evolutionary theory "pleiotropy" also postulates aging as a price for facilitating the reproduction in the early life stage of an organism. For investigating the association between sexual reproduction and aging, a sexual phase-inducible organism in a laboratory would be suitable. One of such organisms is hydra, a genus of Cnidaria. Asexual hydra has been considered to be immortal, but there is the possibility that hydra undergoes aging after sexual reproduction. To search for signs of aging in hydra, we studied sexually differentiated Hydra oligactis at the individual and cellular levels. As a result, we found a significant decline in the capacities for food capture, contractile movements, and reproduction. More importantly, we discovered an exponential increase in the mortality rate of the population. These observations suggest that the degenerative process in H. oligactis represents the aging process. Furthermore, we found that the number of germ cells increased, whereas the number of somatic cells concomitantly decreased. The observed change of the cell composition is thus consistent with the "pleiotropy" theory of aging.

Animals↗

Rapid evolution of major histocompatibility complex class I genes in primates generates new disease alleles in humans via hitchhiking diversity.

A plausible explanation for many MHC-linked diseases is lacking. Sequencing of the MHC class I region (coding units or full contigs) in several human and nonhuman primate haplotypes allowed an analysis of single nucleotide variations (SNV) across this entire segment. This diversity was not evenly distributed. It was rather concentrated within two gene-rich clusters. These were each centered, but importantly not limited to, the antigen-presenting HLA-A and HLA-B/-C loci. Rapid evolution of MHC-I alleles, as evidenced by an unusually high number of haplotype-specific (hs) and hypervariable (hv) (which could not be traced to a single species or haplotype) SNVs within the classical MHC-I, seems to have not only hitchhiked alleles within nearby genes, but also hitchhiked deleterious mutations in these same unrelated loci. The overrepresentation of a fraction of these hvSNV (hv1SNV) along with hsSNV, as compared to those that appear to have been maintained throughout primate evolution (trans-species diversity; tsSNV; included within hv2SNV) tends to establish that the majority of the MHC polymorphism is de novo (species specific). This is most likely reminiscent of the fact that these hsSNV and hv1SNV have been selected in adaptation to the constantly evolving microbial antigenic repertoire.

Alleles↗

magp4 gene may contribute to the diversification of cichlid morphs and their speciation.

Lake Victoria harbors more than 300 species of cichlid fish, which are adapted to a variety of ecological niches with various morphological species-specific features. However, it is believed that these species arose explosively within the last 14,000 years and transcripts among Lake Victoria cichlid species are almost identical in sequence. These data prompted us to develop a DNA chip assay to compare patterns of gene expression among cichlid species. We prepared a DNA chip spotted with 6240 elements derived from cichlid expressed sequence tag (EST) clones and successfully characterized gene expression differences between the cichlid species Haplochromis chilotes and Haplochromis sp. "rockkribensis". We identified 14 transcripts that were differentially expressed between these species at an early developmental stage, 15 days post-fertilization (dpf), and several were further analyzed using quantitative real-time PCR (qPCR). One of these differentially expressed transcripts was a homolog of microfibril-associated glycoprotein 4 (magp4), a putative causative gene for the human inherited disease, Smith-Magenis syndrome (SMS), for which facial defects are among the phenotypic features. Further analysis of magp4 expression showed that magp4 was expressed in the jaw portion of cichlid fry and that expression profiles between Haplochromis chilotes and Haplochromis sp. "rockkribensis" differed during development. These data suggest that the differential expression of a gene associated with human cranial morphogenesis may be involved in the diversification of cichlid jaw morphs.

Animals↗

The Rice Annotation Project Database (RAP-DB): hub for Oryza sativa ssp. japonica genome information.

With the completion of the rice genome sequencing, a standardized annotation is necessary so that the information from the genome sequence can be fully utilized in understanding the biology of rice and other cereal crops. An annotation jamboree was held in Japan with the aim of annotating and manually curating all the genes in the rice genome. Here we present the Rice Annotation Project Database (RAP-DB), which has been developed to provide access to the annotation data. The RAP-DB has two different types of annotation viewers, BLAST and BLAT search, and other useful features. By connecting the annotations to other rice genomics data, such as full-length cDNAs and Tos17 mutant lines, the RAP-DB serves as a hub for rice genomics. All of the resources can be accessed through http://rapdb.lab.nig.ac.jp/.

Databases, Nucleic Acid↗

Phylogeny of the sex-determining gene Sex-lethal in insects.

The Sex-lethal (SXL) protein belongs to the family of RNA-binding proteins and is involved in the regulation of pre-mRNA splicing. SXL has undergone an obvious change of function during the evolution of the insect clade. The gene has acquired a pivotal role in the sex-determining pathway of Drosophila, although it does not act as a sex determiner in non-drosophilids. We collected SXL sequences of insect species ranging from the pea aphid (Acyrtho siphom pisum) to Drosophila melanogaster by searching published articles, sequencing cDNAs, and exploiting homology searches in public EST and whole-genome databases. The SXL protein has moderately conserved N- and C-terminal regions and a well-conserved central region including 2 RNA recognition motifs. Our phylogenetic analysis shows that a single orthologue of the Drosophila Sex-lethal (Sxl) gene is present in the genomes of the malaria mosquito Anopheles gambiae, the honeybee Apis mellifera, the silkworm Bombyx mori, and the red flour beetle Tribolium castaneum. The D. melanogaster, D. erecta, and D. pseudoobscura genomes, however, contain 2 paralogous genes, Sxl and CG3056, which are orthologous to the Anopheles, Apis, Bombyx, and Tribolium Sxl. Hence, a duplication in the fly clade generated Sxl and CG3056. Our hypothesis maintains that one of the genes, Sxl, adopted the new function of sex determiner in Drosophila, whereas the other, CG3056, continued to serve some or all of the yet-unknown ancestral functions.

Amino Acid Sequence↗

Molecular cloning and chromosomal localization of the Bombyx Sex-lethal gene.

We cloned Bm-Sxl, an orthologue of the Drosophila melanogaster Sex-lethal (Sxl) gene from embryos of Bombyx mori. The full-length cDNAs were of 2 sizes, 1528 and 1339 bp, and were named Bm-Sxl-L and Bm-Sxl-S, respectively. Bm-Sxl-L consists of 8 exons and spans more than 20 kb of genomic DNA. The open reading frame (ORF) codes for a protein 336 amino acids in length. Bm-Sxl-S is a splice variant that lacks the second exon. This creates a new translation start 138 nucleotides downstream and an ORF that codes for 46 amino acids fewer at the N-terminus. Linkage analysis using an F2 panel mapped Bm-Sxl to linkage group 16 at 69.8 cM. We isolated 2 BACs that include the Bm-Sxl gene. With BAC-FISH we located Bm-Sxl cytogenetically on the chromosome corresponding to linkage group 16 (LG16) at position >68.8 cM.

Alternative Splicing↗

Comparative genome analyses of nervous system-specific genes.

To elucidate the evolutionary process of the nervous system (NS) in metazoa, we examined the relationship between human genes specifically expressed in the NS (NS-specific genes) and the time of their evolutionary emergence. We obtained 255 human NS-specific genes from the gene expression data of the human full-length cDNA annotation invitational (H-invitational) database. To determine when these genes emerged for the first time during evolution, we searched for orthologues of the 255 NS-specific genes in 13 species (excluding human) by homology searches against their complete genome sequences. We found that 14% of the NS-specific orthologous genes had already emerged before the divergence between yeast and human. This finding suggests that a common ancestor, which should have no nervous system, already possessed a portion of the genes homologous to human NS-specific genes, implying that 14% of the NS-specific genes should have changed differentially their original functions during evolution. If this is the case, then the remaining 86% of the 255 NS-specific human genes have newly emerged during evolution. In particular, we found that the largest portion (24%) of the 255 NS-specific genes had emerged after divergence of urochordata and human but before divergence of fishes and human. These results suggest that the main cause of the NS evolution was the addition of new genes which took place most actively just before or at the evolutionary emergence of vertebrates.

Amino Acid Sequence↗

Evolution of metabolic networks by gain and loss of enzymatic reaction in eukaryotes.

The metabolic network is composed of enzymatic reactions (ERs) in which one or more enzymes catalyze the reaction of pertinent substrates. Since metabolism is a basal system for maintaining life of all organisms, any change in the metabolic networks must greatly affect organismic evolution. The aim of this study is to examine how often gains and losses of ER have occurred during the evolution of metabolic networks in eukaryotes and how these evolutionary events have affected phenotypic traits of organisms. In this study, we conducted comparative studies of 751 ERs in the metabolic networks of 6 eukaryotic species whose complete genome sequences were determined. As a result, we found that a total of 804 gains and losses of ERs had occurred in the evolutionary diversification of metabolic networks in different lineages. Moreover, the vertebrate lineage, after the separation from Drosophila melanogaster, showed a remarkable increase in the number of ER gains compared with ER losses. In particular, 41% of the ER gains were predominantly involved with lipids and complex lipid metabolism. Because some products of these two metabolisms function as hormones, we concluded that the ER gain of these two metabolisms accelerated the development of hormonal signal transduction for the elaborate regulation of physiological systems during vertebrate evolution.

Amino Acid Sequence↗

Development of a spot reliability evaluation score for DNA microarrays.

We developed a reliability index named SRED (Spot Reliability Evaluation Score for DNA microarrays) that represents the probability that the calibrated gene expression level from a DNA microarray would be less than a factor of 2 different from that of quantitative real-time polymerase chain reaction assays whose dynamic quantification range is treated statistically to be similar to that of the DNA microarray. To define the SRED score, two parameters, the reproducibility of measurement value and the relative expression value were selected from nine candidate parameters. The SRED score supplies the probability that the expression level in each spot of a microarray is less than a certain-fold different compared to other expression profiling data, such as QRT-PCR. This score was applied to approximately 1,500,000 points of the expression profile in the RIKEN Expression Array Database.

Algorithms↗

Estimation of ancestral gene set of bilaterian animals and its implication to dynamic change of gene content in bilaterian evolution.

To understand the process of bilaterian evolution, we estimated ancestral gene sets at the split of plant-animal-fungi and the divergence of bilaterian animals and from 1,236,790 non-redundant genes. We, then, examined how the numbers of the gene clusters have changed since the split. As a result, we estimated the numbers of gene clusters in the ancestral gene sets of plant-animal-fungi and bilaterian animals to be at least 2469 and 6577, respectively. Thus, we found a 2.7-fold increase in the number of gene clusters during the period from the evolutionary split of plant-animal-fungi to the divergence of bilaterian animals. Moreover, when we compared these numbers of ancestral gene clusters with those of extant animals such as the nematode, fly, mouse and human, we found that the extant bilaterian animals have retained more than 3500 gene clusters of the ancestral gene set, and have lost more than 1600 gene clusters. It suggests that these processes of genomic diversification provided bilaterian animals with molecular basis for species diversity.

Animals↗

The Human Anatomic Gene Expression Library (H-ANGEL), the H-Inv integrative display of human gene expression across disparate technologies and platforms.

The Human Anatomic Gene Expression Library (H-ANGEL) is a resource for information concerning the anatomical distribution and expression of human gene transcripts. The tool contains protein expression data from multiple platforms that has been associated with both manually annotated full-length cDNAs from H-InvDB and RefSeq sequences. Of the H-Inv predicted genes, 18 897 have associated expression data generated by at least one platform. H-ANGEL utilizes categorized mRNA expression data from both publicly available and proprietary sources. It incorporates data generated by three types of methods from seven different platforms. The data are provided to the user in the form of a web-based viewer with numerous query options. H-ANGEL is updated with each new release of cDNA and genome sequence build. In future editions, we will incorporate the capability for expression data updates from existing and new platforms. H-ANGEL is accessible at http://www.jbirc.aist.go.jp/hinv/h-angel/.

Database Management Systems↗

Structural and functional analysis of a 0.5-Mb chicken region orthologous to the imprinted mammalian Ascl2/Mash2-Igf2-H19 region.

Previous studies revealed that Igf2 and Mpr/Igf2r are imprinted in eutherian mammals and marsupials but not in monotremes or birds. Igf2 lies in a large imprinted cluster in eutherians, and its imprinting is regulated by long-range mechanisms. As a step to understand how the imprinted cluster evolved, we have determined a 490-kb chicken sequence containing the orthologs of mammalian Ascl2/Mash2, Ins2 and Igf2. We found that most of the genes in this region are conserved between chickens and mammals, maintaining the same transcriptional polarities and exon-intron structures. However, H19, an imprinted noncoding transcript, was absent from the chicken sequence. Chicken ASCL2/CASH4 and INS, the orthologs of the imprinted mammalian genes, showed biallelic expression, further supporting the notion that imprinting evolved after the divergence of mammals and birds. The H19 imprinting center and many of the local regulatory elements identified in mammals were not found in chickens. Also, a large segment of tandem repeats and retroelements identified between the two imprinted subdomains in mice was not found in chickens. Our findings show that the imprinted genes were clustered before the emergence of imprinting and that the elements associated with imprinting probably evolved after the divergence of mammals and birds.

Amino Acid Sequence↗

Detection of apoptosis during planarian regeneration by the expression of apoptosis-related genes and TUNEL assay.

Apoptosis is a tightly organized cell death process that plays a crucial role in metazoan development, but it has not yet been revealed whether apoptotic events are involved in the process of regeneration. Here, we tried to detect apoptotic cells during planarian regeneration using the TdT-mediated dUTP nick-end labeling (TUNEL) assay as well as the expression of apoptosis-related genes. Three novel cDNAs were isolated from a planarian cDNA library and shown to be closely related to other metazoan caspases at the amino acid sequence level. One of these cDNAs, Caspase-like gene 3 (DjClg3), was expressed primarily in apoptotic cells by double detections with the TUNEL assay. Whole mount in situ studies indicated that DjClg3 was expressed in the cells of the mesenchymal space and also around the pharynx of the intact body. Its expression in the regenerating head piece was seen in the blastema and less significantly in the brain, while in the regenerating tail piece, DjClg3 expression was detected uniformly throughout the entire region. In parallel experiments, we performed in situ TUNEL assays to localize the regions where cell death occurred during regeneration and comparable results to the DjClg3 expression patterns were obtained. This is the first report to show that planarians have apoptosis-related genes and the results suggest that the apoptotic mechanism probably takes place to a large extent in normal intact worms as well as during their regeneration. We hypothesize that the presence of apoptosis in planarians may have a role in controlling cell numbers, eliminating unnecessary tissues or cells and remodeling the old tissues of regenerating body parts.

Amino Acid Sequence↗

The evolutionary origin of long-crowing chicken: its evolutionary relationship with fighting cocks disclosed by the mtDNA sequence analysis.

Chickens with exceptionally long crow are often favored all over the world, and connoisseur breeders have bred certain types of chicken exclusively for this trait. In Japan, three chicken varieties have been specifically bred to develop an exceptionally long crow of over 15 s. Although these three long-crowing chickens, Naganakidori, are honored as heritage varieties of Japan, the domestication process and genealogical origin of long-crowing chickens remain unclear. The purpose of this study is to clarify these issues using nucleotide sequences of the mitochondrial DNA D-loop region. Blood samples from a total of nine long-crowing chickens and 74 chickens from 11 Japanese native varieties were collected. DNA sequence data of two Junglefowl species were also collected from the International DNA database (DDBJ /EMBL/GenBank) for use as the outgroup. A phylogenetic tree was then constructed revealing that all three Naganakidori varieties were monophyletic and originated from a fighting cock, a Shamo, for cockfighting. These results suggest that these three long-crowing chickens share a common origin in spite of their conspicuously different characters, and that human cultures favoring long-crowing chickens might have been preceded by a tradition of cockfighting. Moreover, these long-crowing varieties first separated from the fighting cocks of Okinawa, which is geographically closer to Southern China and Indochina than Mainland Japan (Honshu/Kyushu). This implies that Japanese long-crowing chickens were first brought to Mainland Japan as fighting cocks from the surrounding regions of Southern China or Indochina and through Okinawa.

Aggression↗

Evolutionary process of amino acid biosynthesis in Corynebacterium at the whole genome level.

Corynebacterium glutamicum, which is the closest relative of Corynebacterium efficiens, is widely used for the large scale production of many kinds of amino acids, particularly glutamic acid and lysine, by fermentation. Corynebacterium diphtheriae, which is well known as a human pathogen, is also closely related to these two species of Corynebacteria, but it lacks such productivity of amino acids. It is an important and interesting question to ask how those closely related bacterial species have undergone such significant functional differentiation in amino acid biosynthesis. The main purpose of the present study is to clarify the evolutionary process of functional differentiation among the three species of Corynebacteria by conducting a comparative analysis of genome sequences. When Mycobacterium and Streptomyces were used as out groups, our comparative study suggested that the common ancestor of Corynebacteria already possessed almost all of the gene sets necessary for amino acid production. However, C. diphtheriae was found to have lost the genes responsible for amino acid production. Moreover, we found that the common ancestor of C. efficiens and C. glutamicum have acquired some of genes responsible for amino acid production by horizontal gene transfer. Thus, we conclude that the evolutionary events of gene loss and horizontal gene transfer must have been responsible for functional differentiation in amino acid biosynthesis of the three species of Corynebacteria.

Amino Acids↗