Shinichi Morishita

The medaka draft genome and insights into vertebrate genome evolution

Teleosts comprise more than half of all vertebrate species and have adapted to a variety of marine and freshwater habitats. Their genome evolution and diversification are important subjects for the understanding of vertebrate evolution. Although draft genome sequences of two pufferfishes have been published, analysis of more fish genomes is desirable. Here we report a high-quality draft genome sequence of a small egg-laying freshwater teleost, medaka (Oryzias latipes). Medaka is native to East Asia and an excellent model system for a wide range of biology, including ecotoxicology, carcinogenesis, sex determination and developmental genetics. In the assembled medaka genome (700 megabases), which is less than half of the zebrafish genome, we predicted 20,141 genes, including ∼2,900 new genes, using 5′-end serial analysis of gene expression tag information. We found single nucleotide polymorphisms (SNPs) at an average rate of 3.42% between the two inbred strains derived from two regional populations; this is the highest SNP rate seen in any vertebrate species. Analyses based on the dense SNP information show a strict genetic separation of 4 million years (Myr) between the two populations, and suggest that differential selective pressures acted on specific gene categories. Four-way comparisons with the human, pufferfish (Tetraodon), zebrafish and medaka genomes revealed that eight major interchromosomal rearrangements took place in a remarkably short period of ∼50 Myr after the whole-genome duplication event in the teleost ancestor and afterwards, intriguingly, the medaka genome preserved its ancestral karyotype for more than 300 Myr.

A large-scale full-length cDNA analysis to explore the budding yeast transcriptome

We performed a large-scale cDNA analysis to explore the transcriptome of the budding yeast Saccharomyces cerevisiae. We sequenced two cDNA libraries, one from the cells exponentially growing in a minimal medium and the other from meiotic cells. Both libraries were generated by using a vector-capping method that allows the accurate mapping of transcription start sites (TSSs). Consequently, we identified 11,575 TSSs associated with 3,638 annotated genomic features, including 3,599 ORFs, to suggest that most yeast genes have two or more TSSs. In addition, we identified 45 previously undescribed introns, including those affecting current ORF annotations and those spliced alternatively. Furthermore, the analysis revealed 667 transcription units in the intergenic regions and transcripts derived from antisense strands of 367 known features. We also found that 348 ORFs carry TSSs in their 3′-halves to generate sense transcripts starting from inside the ORFs. These results indicate that the budding yeast transcriptome is considerably more complex than previously thought, and it shares many recently revealed characteristics with the transcriptomes of mammals and other higher eukaryotes. Thus, the genome-wide active transcription that generates novel classes of transcripts appears to be an intrinsic feature of the eukaryotic cells. The budding yeast will serve as a versatile model for the studies on these aspects of transcriptome, and the full-length cDNA clones can function as an invaluable resource in such studies.

Mining Optimized Association Rules for Numeric Attributes

Given a huge database, we address the problem of finding association rules for numeric attributes, such as(Balance?I)?(CardLoan=yes),which implies that bank customers whose balances fall in a rangeIare likely to use card loan with a probability greater thanp. The above rule is interesting only if the rangeIhas some special feature with respect to the interrelation betweenBalanceandCardLoan. It is required that the number of customers whose balances are contained inI(called thesupportofI) is sufficient and also that the probabilitypof the conditionCardLoan=yesbeing met (called theconfidence ratio) be much higher than the average probability of the condition over all the data. Our goal is to realize a system that finds such appropriate ranges automatically. We mainly focus on computing twooptimized ranges: one that maximizes the support on the condition that the confidence ratio is at least a given threshold value, and another that maximizes the confidence ratio on the condition that the support is at least a given threshold number. Using techniques from computational geometry, we present novel algorithms that compute the optimized ranges in linear time if the data are sorted. Since sorting data with respect to each numeric attribute is expensive in the case of huge databases that occupy much more space than the main memory, we instead apply randomized bucketing as the preprocessing method and thus obtain an efficient rule-finding system. Tests show that our implementation is fast not only in theory but also in practice. The efficiency of our algorithm enables us to compute optimized rules for all combinations of hundreds of numeric and Boolean attributes in a reasonable time.

Transversing itemset lattices with statistical metric pruning

We study how to efficiently compute significant association rules according to common statistical measures such as a chi-squared value or correlation coefficient. For this purpose, one might consider to use of the Apriori algorithm, but the algorithm needs major conversion, because none of these statistical metrics are anti-monotone, and the use of higher support for reducing the search space cannot guarantee solutions in its the search space. We here present a method of estimating a tight upper bound on the statistical metric associated with any superset of an itemset, as well as the novel use of the resulting information of upper bounds to prune unproductive supersets while traversing itemset lattices. Experimental tests demonstrate the efficiency of this method.

Diverse transcriptional initiation revealed by fine, large‐scale mapping of mRNA start sites

Determination of the mRNA start site is the first step in identifying the promoter region, which is of key importance for transcriptional regulation of gene expression. The ‘oligo‐capping’ method enabled us to introduce a sequence tag to the first base of an mRNA by replacing the cap structure of the mRNA. Using cDNA libraries made from oligo‐capped mRNAs, we could identify the transcriptional start site of an individual mRNA just by sequencing the 5′‐end of the cDNA. The fine mapping of transcriptional start sites was performed for 5880 mRNAs in 276 human genes. Contrary to our expectations, the majority of the genes showed a diverse distribution of transcriptional start sites. They were distributed over 61.7 bp with a standard deviation of 19.5. Our finding may reflect the dynamic nature of transcriptional initiation events of human genes in vivo.

dsCheck: highly sensitive off-target search software for double-stranded RNA-mediated RNA interference

Off-target effects are one of the most serious problems in RNA interference (RNAi). Here, we present dsCheck (), web-based online software for estimating off-target effects caused by the long double-stranded RNA (dsRNA) used in RNAi studies. In the biochemical process of RNAi, the long dsRNA is cleaved by Dicer into short-interfering RNA (siRNA) cocktails. The software simulates this process and investigates individual 19 nt substrings of the long dsRNA. Subsequently, the software promptly enumerates a list of potential off-target gene candidates based on the order of off-target effects using its novel algorithm, which significantly improves both the efficiency and the sensitivity of the homology search. The website not only provides a rigorous off-target search to verify previously designed dsRNA sequences but also presents ‘off-target minimized’ dsRNA design, which is essential for reliable experiments in RNAi-based functional genomics.

Chromatin-associated periodicity in genetic variation downstream of transcriptional start sites

Might DNA sequence variation reflect germline genetic activity and underlying chromatin structure? We investigated this question using medaka (Japanese killifish, Oryzias latipes), by comparing the genomic sequences of two strains (Hd-rR and HNI) and by mapping ∼37.3 million nucleosome cores from Hd-rR blastulae and 11,654 representative transcription start sites from six embryonic stages. We observed a distinctive ∼200–base pair (bp) periodic pattern of genetic variation downstream of transcription start sites; the rate of insertions and deletions longer than 1 bp peaked at positions of approximately +200, +400, and +600 bp, whereas the point mutation rate showed corresponding valleys. This ∼200-bp periodicity was correlated with the chromatin structure, with nucleosome occupancy minimized at positions 0, +200, +400, and +600 bp. These data exemplify the potential for genetic activity (transcription) and chromatin structure to contribute to molding the DNA sequence on an evolutionary time scale.

5′-end SAGE for the analysis of transcriptional start sites

Identification of the mRNA start site is essential in establishing the full-length cDNA sequence of a gene and analyzing its promoter region, which regulates gene expression. Here we describe the development of a 5′-end serial analysis of gene expression (5′ SAGE) that can be used to globally identify transcriptional start sites and the frequency of individual mRNAs. Of the 25,684 5′ SAGE tags in the HEK293 human cell library, 19,893 matched to the human genome. Among 15,448 tags in one locus of the genome, 85.8%–96.1% of the 5′ SAGE tags were assigned within −500 to +200 nt of mRNA start sites using the RefSeq, UniGene and DBTSS databases. This technique should facilitate 5′-end transcriptome analysis in a variety of cells and tissues.

KDD Cup 2001 report

This paper presents results and lessons from KDD Cup 2001. KDD Cup 2001 focused on mining biological databases. It involved three cutting-edge tasks related to drug design and genomics.

Comparative analysis of chimpanzee and human y chromosomes unveils complex evolutionary pathway

The mammalian Y chromosome has unique characteristics compared with the autosomes or X chromosomes. Here we report the finished sequence of the chimpanzee Y chromosome (PTRY), including 271 kb of the Y-specific pseudoautosomal region 1 and 12.7 Mb of the male-specific region of the Y chromosome. Greater sequence divergence between the human Y chromosome (HSAY) and PTRY (1.78%) than between their respective whole genomes (1.23%) confirmed the accelerated evolutionary rate of the Y chromosome. Each of the 19 PTRY protein-coding genes analyzed had at least one nonsynonymous substitution, and 11 genes had higher nonsynonymous substitution rates than synonymous ones, suggesting relaxation of selective constraint, positive selection or both. We also identified lineage-specific changes, including deletion of a 200-kb fragment from the pericentromeric region of HSAY, expansion of young Alu families in HSAY and accumulation of young L1 elements and long terminal repeat retrotransposons in PTRY. Reconstruction of the common ancestral Y chromosome reflects the dynamic changes in our genomes in the 5–6 million years since speciation.