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S Kusakabe

Publications and source records attributed to S Kusakabe.

At least 19 recordsLinked to original sources
PubMed2000

The genetic structure of the Raleigh natural population of Drosophila melanogaster revisited.

The Raleigh natural population of Drosophila melanogaster was reanalyzed with special attention to possible dysgenic effects during the extraction of chromosomes. About 600 second chromosomes were extracted from the Raleigh natural population, half in the cytoplasm of wild-caught females (native genetic background) and half in the cytoplasm of the laboratory line, C160(In(2LR)SM1, Cy/In(2LR)bw(V1)) (foreign genetic background). We could not find significant differences between the two extraction schemes in the frequency of lethal second chromosomes (Q = 0.252 for the lines with the negative genetic background vs. 0.231 for the lines with the foreign genetic background) or in the homozygous detrimental (D) and lethal (L) loads (D = 0.210 vs. 0.251; L = 0.287 vs. 0.264). The effective size of the population was estimated to be approximately 19,000, based on the allelism rate of lethal-bearing chromosomes. The homozygous load markedly decreased in the 15 years since a previous study of the same population.

Animals↗
PubMed1998

Phylogenetic positions of insectivora in eutheria inferred from mitochondrial cytochrome c oxidase subunit II gene.

For the elucidation of the phylogenetic position of insectivora in eutheria, we have sequenced the cytochrome c oxidase subunit II (COII) gene of mitochondria for three insectivoran species [musk screw (Suncus murinus), shrew mole (Urotrichus talpoides), Japanese mole (Mogera wogura)] and analyzed these amino acid sequences with neighbor-joining (NJ) method and maximum likelihood (ML) method. NJ analysis shows polyphyly of Insectivora and Chiroptera. Assuming that each of Primates, Ferungulata, Chiroptera, Insectivora and Rodentia is a monophyletic group, ML analysis suggests that Chiroptera is a sister group of Insectivora and that Ferungulata is the closest outgroup to the (Insectivora and Chiroptera) clade.

Animals↗
PubMed1993

Distribution of polymorphic gene duplication at the Gpdh locus in natural populations of Drosophila melanogaster.

The glycerol-3-phosphate dehydrogenase (GPDH, E.C. 1.1.1.8) gene of Drosophila melanogaster contains a tandem duplication of a 4.5-kb-long DNA fragment. Survey of the Gpdh gene region by the Southern blot analysis revealed the following features of this gene duplication: (1) The duplication was not observed in chromosome lines that carry In(2L)t, a cosmopolitan chromosomal inversion in this species. The duplication and the inversion are in linkage disequilibrium. (2) The duplication is polymorphic in the Japan and US natural populations examined. Its frequency is 0.26 on an average in In(2L)t-free chromosomes. (3) Triplication is absent or has not become frequent in the populations surveyed. Possible evolutionary factors of this duplication polymorphism are discussed.

Alleles↗
PubMed1993

Spontaneous mutation rates in null and band-morph mutations of enzyme loci in Drosophila melanogaster.

Spontaneous mutations were accumulated for a total of 1,678,388 allele-generations in several hundred replicate second chromosome lines. These lines were compared of a Cy chromosome and one of three different lethal bearing chromosomes from natural populations. Electrophoretic mobility and/or activity change was screened for seven enzyme loci. Forty-four null mutations were detected, but no band-morph mutations were observed. There is significant variation in null mutation rate among enzyme loci and among lethal chromosomes. This may be attributable to differences in the distribution of transposable elements among the lines. The band-morph mutation rate is pooled with previous results and becomes 7.48 x 10(-7) with a 95% confidence limit of 2.04 x 10(-7) to 1.91 x 10(-6) per locus per generation. Similarly, the pooled null mutation rate becomes 1.30 x 10(-5) with 95% confidence limit of 1.15 x 10(-5) to 1.52 x 10(-5) per locus per generation.

Animals↗
PubMed1992

Spontaneous mutations affecting glycerol-3-phosphate dehydrogenase enzyme activity in Drosophila melanogaster.

Significant genetic variance in glycerol-3-phosphate dehydrogenase (GPDH) activity was observed between chromosome lines of Drosophila melanogaster that had each accumulated spontaneous mutations for approximately 300 generations. No restriction map variation was found in a 26-kb region surrounding the entire Gpdh gene. The restriction analysis used is capable of detecting insertions/deletions larger than 0.05 kb. The survey would also detect chromosomal recombinations that include the entire Gpdh coding region. Therefore, if the spontaneous mutations that affected the enzyme activity are located inside the Gpdh gene region, then they are base pair substitutions or structural changes that are smaller than the limit in resolution described above.

Alcohol Dehydrogenase↗
PubMed1991

A method for detecting effect of beneficial mutations in natural populations of Drosophila melanogaster.

An experimental method is proposed for detecting the effects of positive natural selection on DNA polymorphisms. Since beneficial mutations are expected to increase in frequency faster than neutral mutations, variants which have reached high frequencies in a relatively short period could be linked to some beneficial mutation. D. melanogaster has a cosmopolitan polymorphic inversion -In(2L)t- whose age in some local populations has been estimated. Setting the age of In(2L)t as the upper limit for the age of variants, we searched for variants whose frequencies were possibly influenced by positive natural selection. We detected a single candidate whose frequency and distribution met the requirements imposed by our method.

Alleles↗
PubMed1991

The genetic structure of natural populations of Drosophila melanogaster. XXII. Comparative study of DNA polymorphisms in northern and southern natural populations.

Restriction map variation in four gene regions (Adh, Amy, Pu and Gpdh) was surveyed for 86 second chromosomes from northern (Aomori) and southern (Ogasawara) Japanese populations of Drosophila melanogaster (43 chromosomes from each population). The regions examined cover a total of 62 kilobases. Estimates of nucleotide diversity (pi) were approximately constant across the gene regions and populations examined. The distribution of restriction site polymorphisms was compatible with the expectation from the neutral mutation-random genetic drift hypothesis, but insertion/deletion polymorphisms were not consistent with it. While the two populations shared a majority of restriction site polymorphisms, frequencies of individual restriction site variants were significantly different between the two populations at 7 out of 35 segregating sites. In addition, an insertion in the Amy region was found in 15 chromosomes from the Ogasawara sample but absent in the Aomori sample. A considerable difference was observed in the number of rare insertions and deletions between the two populations. The numbers of aberrations uniquely represented were 16 in the Ogasawara sample and only 3 in the Aomori sample. These findings suggest that the two populations were differentiated from each other to some degree by means of random genetic drift and/or other factors.

Alcohol Dehydrogenase↗
PubMed1991

Comparative study of P element activity in two natural populations of Drosophila melanogaster.

Population structure concerning P element activity was investigated in two natural Drosophila populations. These populations are very different as far as in the viability spectrum is concerned. In one population, the Raleigh, United States population, genetic loads related to viability have been kept at a fairly high level. In the other population, the Nagasaki, Japan, population, the genetic loads tend to be stable at very low levels. In the Raleigh population it is estimated that on the average 4 copies of intact P elements that possess transposase activity exist in the genome. On the other hand only 0.7 complete copies are estimated to exist in the genome of the Nagasaki population. Heterogeneity in the P element copy number and significant positive linkage disequilibrium among occupied sites were detected in the Raleigh population. Our results, with some evidences which indicate that high mutation rate was caused by the P element, suggests that the large genetic loads in the Raleigh population are caused by the rapid invasion of P element in this population.

Animals↗
PubMed1990

Gene duplication and concerted evolution of the GPDH locus in natural populations of Drosophila melanogaster.

The sn-glycerol-3-phosphate dehydrogenase (GPDH, EC 1, 1, 1, 8) locus of Drosophila melanogaster is polymorphic with respect to the number of tandemly duplicated genes in natural populations. The duplicated genes were cloned and the nucleotide sequences were determined. The duplication deletes both the first and second exons and has a size of 4500 b.p. The fact that there is no sequence variation at the junction point of the duplicated units among strains suggests a single origin for the duplication event. Comparison of the nucleotide sequences among the duplicates indicates that the frequent transfer of genetic information occurs from one to the other of the duplicates on the same chromosome either by gene conversion or by unequal crossing over. Because the GPDH duplication is partial and therefore a kind of pseudogene, the observed polymorphism of the number of tandemly duplicated GPDH genes appears to have been driven mainly by random genetic drift.

Animals↗
PubMed1990

The rare inversion with a P element at the breakpoint maintained in a natural population of Drosophila melanogaster.

We found a rare inversion which is superimposed on the In(2R) NS chromosome. It has been maintained for several years in the Raleigh, N.C., USA population and possibly for more than ten years in the southern populations of the USA. Both the breakpoint of the rare inversion and the corresponding site of the In(2R) NS chromosome have P elements and therefore, the rare inversion might be induced by P element activity in a natural population.

Animals↗
PubMed1989

Sequence, structure and evolution of the gene coding for sn-glycerol-3-phosphate dehydrogenase in Drosophila melanogaster.

We present the complete nucleotide and deduced amino acid sequence for the gene encoding Drosophila sn-glycerol-3-phosphate dehydrogenase. A transcription unit of 5kb was identified which is composed of eight protein encoding exons. Three classes of transcripts were shown to differ only in the 3'-end and to code for three protein isoforms each with a different C-terminal amino acid sequence. Each transcript is shown to arise through the differential expression of three isotype-specific exons at the 3'-end of the gene by a developmentally regulated process of 3'-end formation and alternate splicing pathways of the pre-mRNA. In contrast, the 5'-end of the gene is simple in structure and each mRNA is transcribed from the same promoter sequence. A comparison of the organization of the Drosophila and murine genes and the primary amino acid sequence between a total of four species indicates that the GPDH gene-enzyme system is highly conserved and is evolving slowly.

Amino Acid Sequence↗
PubMed1989

Structure of the rat alpha 2-macroglobulin-coding gene.

Rat alpha 2-macroglobulin (alpha 2M) is an acute-phase protein, i.e., produced upon tissue inflammation. Genomic DNA clones covering the entire sequence of the alpha 2M gene were isolated and characterized by restriction mapping. Southern blotting and (partial) DNA sequencing. The rat alpha 2M gene is approx. 50 kb in length and consists of 36 exons ranging in size from 21 to 229 bp. Two functional domains, a bait region and a thiol ester site, are encoded by the exon 18 and 24, respectively. Several possible regulatory signals such as a TPA-inducible enhancer core, an identifier sequence, purine-pyrimidine alternative stretches and viral enhancer core sequences were identified. Several genomic DNA clones which cross-hybridized with the alpha 2M cDNA probe were also identified. Sequence analysis showed that they possessed sequences identical to a part of the rat alpha 1-inhibitor III cDNA and that they had a strikingly similar exon organization to the alpha 2M gene.

Animals↗
PubMed1989

Polymorphism for the number of tandemly multiplicated glycerol-3-phosphate dehydrogenase genes in Drosophila melanogaster.

A 26-kilobase-pair region encompassing the sn-glycerol-3-phosphate dehydrogenase (sn-glycerol-3-phosphate:NAD+ 2-oxidoreductase, EC 1.1.1.8) locus in Drosophila melanogaster from two natural populations in Japan was surveyed by restriction mapping. Both tandem duplications and triplications in this region were found in both populations. Detailed analysis of 86 chromosome 2 lines revealed restriction site and allozyme polymorphisms in the transcriptional unit: two restriction sites and the allozymes [fast (F) or slow (S)] were polymorphic among both duplication-bearing chromosomes and those carrying the standard sequence. This finding suggests recurrent recombination and/or gene conversion in this 5-kilobase-pair region. The differences observed for restriction site and allozyme haplotypes among the triplicated sequence both within and between populations, together with the distribution in natural populations, suggest a relatively recent ancestry of the triplication events and an independent origin in respective populations. Such events may represent the process of the formation of multigene families [compare Ohta, T. (1987) Genetics 115, 207-213]. Finally, the evolution of this type of polymorphism is discussed.

Animals↗
PubMed1988

Antibacterial and pharmacokinetic properties of M14659, a new injectable semisynthetic cephalosporin.

In vitro and in vivo antibacterial activities and pharmacokinetics of M14659 were investigated. In vitro activity of M14659 was superior to that of ceftazidime against Staphylococcus aureus. Against Gram-negative bacteria except Pseudomonas aeruginosa, its activity was almost equal to that of ceftazidime. M14659 was more active against P. aeruginosa including multi-drug resistant strains than cefsulodin, cefoperazone or ceftazidime. Affinities of M14659 for penicillin-binding proteins (PBPs) of Escherichia coli and P. aeruginosa were 2 to 14 times higher for PBP-1A and PBP-1B than found for ceftazidime, and almost the same for PBP-3. In vivo activity of M14659 against S. aureus was superior to that of cefamandole, cefotaxime or ceftazidime. Against Gram-negative bacteria including P. aeruginosa, M14659 was 2 to 220 times more active than cefotaxime or ceftazidime. Plasma half-life of M14659 in mice was about 3 times longer than that of ceftazidime. M14659 administered intravenously to mice was mainly excreted in urine without metabolism, and its recovery rate was almost equal to that of ceftazidime.

Animals↗
PubMed1987

The genetic structure of natural populations of Drosophila melanogaster. XX. Comparison of genotype-environment interaction in viability between a northern and a southern population.

In order to examine the operation of diversifying selection as the maintenance mechanism of excessive additive genetic variance for viability in southern populations in comparison with northern populations of Drosophila melanogaster, two sets of experiments were conducted using second chromosomes extracted from the Ogasawara population (a southern population in Japan) and from the Aomori population (a northern population in Japan). Chromosomal homozygote and heterozygote viabilities were estimated in eight kinds of artificially produced breeding environments. The main findings in the present investigation are as follows: (1) Significant genotype-environment interaction was observed using chromosomes extracted from the Ogasawara population. Indeed, the estimate of the genotype-environment interaction variance for heterozygotes was significantly larger than that of the genotypic variance. On the other hand, when chromosomes sampled from the Aomori population were examined, that interaction variance was significant only for homozygotes and its value was no more than one quarter of that for the chromosomes from the Ogasawara population. (2) The average genetic correlation between any two viabilities of the same lines estimated in the eight kinds of breeding environments for the chromosomes sampled from the Ogasawara population was smaller than that for the chromosomes from the Aomori population both in homozygotes and in heterozygotes, especially in the latter. (3) The stability of heterozygotes over homozygotes against fluctuations of environmental conditions was seen in the chromosomes from the Ogasawara population, but not from the Aomori population. (4) From the excessive genotype-environment interaction variance compared with the genotypic variance in heterozygotes, it was suggested for the chromosomes from the Ogasawara population that the reversal of viability order between homozygotes took place in some environments at the locus level. On the basis of these findings, it is strongly suggested that diversifying selection is operating in a southern population of D. melanogaster on some of the viability polygenes which are probably located outside the structural loci, and the excessive additive genetic variance of viability in southern populations is maintained by this type of selection.

Alleles↗
PubMed1985

Rapid change in mutation rate in a local population of Drosophila melanogaster.

The lethal and detrimental loads per second chromosome rapidly increased from 1968 to 1970 in a local population of Drosophila melanogaster in Japan (lethal load, from about 0.16 to 0.38; detrimental load, from 0.125 to 0.231 [Watanabe, T. K., Watanabe, T. & Oshima, C. (1976) Evolution 30, 109-118]). When the homozygous loads were measured in 1983, the lethal load had decreased to approximately the original amount (0.19) but the detrimental load had stayed high (0.241). The rise and fall of the lethal load can be accounted for by a P-type element that invaded a population with M cytotype, producing a high mutation rate. The mutation rate fell back to the earlier value after the cytotype became P. That the detrimental load did not decrease can be explained by assuming a longer persistence for detrimental mutations in the population. Evidence for a P-type mutator factor is that the mutation rate of the wild-type chromosomes differs between the different cytoplasmic and chromosomal backgrounds, being lower in the background from which the chromosomes were taken.

Animals↗
PubMed1983

Reexamination of diversifying selection of polymorphic allozyme genes by using population cages in Drosophila melanogaster.

Several investigators have reported increased heterozygosity for allozyme markers in laboratory experiments simulating a variable environment and have concluded that there is diversifying selection for allozyme variants. We found no change in heterozygosity in 28 large population cages where temperature, food, and light were each set at three levels, two constant and one variable, in a factorial design. Specifically, there was no change in heterozygosity when 0, 1, 2, or 3 factors were variable. The discrepancy between our negative results and earlier positive ones is that our experiments were started from a large caged population of flies which had been maintained for 6 years-long enough to be very near to linkage equilibrium. We suggest that the earlier results may have been the result of linkage between neutral, or nearly neutral, allozyme markers and selected loci or in some cases, inversions.

Journal Article↗