Molecular characterization and evolution of the repeating units of histone genes in Drosophila americana: coexistence of quartet and quintet units in a genome

Quintet and quartet repeating units of the histone genes in Drosophila americana were cloned and characterized. Nucleotide sequence analysis of the units showed that a 3175 bp unit contained the core histone genes but lacked the H1 gene ('quartet unit') while a 5025 bp unit contained all five histone genes ('quintet unit'). Comparative analysis suggested that these repeating units diverged before the separation of D. americana and D. virilis. Multiple forms of H1 genes, differing by 5.8% of amino acids, were found in D. americana. The genomic organization of the histone gene family in D. americana was found to be very similar to that of D. virilis.


Divergence and heterogeneity of the histone gene repeating units in the Drosophila melanogaster species subgroup.

The repeating units of the histone gene cluster containing the H1, H2A, H2B and H4 genes were amplified by PCR from the Drosophila melanogaster species subgroup, i.e., D. yakuba, D. erecta, D. sechellia, D. mauritiana, D. teissieri and D. orena. The PCR products were cloned and their nucleotide sequences of about 4.6-4.8kbp were determined to elucidate the mechanism of molecular evolution of the histone gene family. The heterogeneity among the histone gene repeating units was 0.6% and 0.7% for D. yakuba and D. sechellia, respectively, indicating the same level of heterogeneity as in the H3 gene region of D. melanogaster. Divergence of the genes among species even in the most closely related ones was much greater than the heterogeneity among family members, indicating a concerted mode of evolution for the histone gene repeating units. Among the species in the D. melanogaster species subgroup, the histone gene regions as well as 3rd codon position of the coding region showed nearly the same GC contents. These results suggested that the previous conclusion on analysis of the H3 gene regions, the gene family evolution in a concerted fashion, holds true for the whole histone gene repeating unit.


Evolution of the GC content of the histone 3 gene in seven Drosophila species.

The molecular evolution of the histone multigene family was studied by cloning and determining the nucleotide sequences of the histone 3 genes in seven Drosophila species, D. takahashii, D. lutescens, D. ficusphila, D. persimilis, D.pseudoobscura, D. americana and D. immigrans. CT repeats, a TATA box and an AGTG motif in the 5' region, and a hairpin loop and purine-rich motifs (CAA(T/G)GAGA) in the 3' region were conserved even in distantly related species. In D. hydei and D.americana, the GC content at the third codon position in the protein coding region was relatively low (49% and 45%), while in D. takahashii and D. lutescens it was relatively high (64% and 65%). The non- significant correlation between the GC contents in the 3' region and at the third codon position as well as the evidence of less constraint in the 3' region suggested that mutational bias may not be the major mechanism responsible for the biased nucleotide change at the third codon position or for codon usage bias.


Molecular evolutionary analysis of a histone gene repeating unit from Drosophila simulans.

A repeating unit of the histone gene cluster from Drosophila simulans containing the H1, H2A, H2B and H4 genes (the H3 gene region has already been analyzed) was cloned and analyzed. A nucleotide sequence of about 4.6 kbp was determined to study the nucleotide divergence and molecular evolution of the histone gene cluster. Comparison of the structure and nucleotide sequence with those of Drosophila melanogaster showed that the four histone genes were located at identical positions and in the same directions. The proportion of different nucleotide sites was 6.3% in total. The amino acid sequence of H1 was divergent, with a 5.1% difference. However, no amino acid change has been observed for the other three histone proteins. Analysis of the GC contents and the base substitution patterns in the two lineages, D. melanogaster and D. simulans, with a common ancestor showed the following. 1) A strong negative correlation was found between the GC content and the nucleotide divergence in the whole repeating unit. 2) The mode of molecular evolution previously found for the H3 gene was also observed for the whole repeating unit of histone genes; the nucleotide substitutions were stationary in the 3' and spacer regions, and there was a directional change of the codon usage to the AT-rich codons. 3) No distinct difference in the mode or pattern of molecular evolution was detected for the histone gene repeating unit in the D. melanogaster and D. simulans lineages. These results suggest that selectional pressure for the coding regions of histones, which eliminate A and T, is less effective in the D. melanogaster and D. simulans lineages than in the other GC-rich species.


Molecular evolution of the histone 3 multigene family in the Drosophila melanogaster species subgroup

Molecular evolution of the histone multigene family was studied by cloning and sequencing regions of the histone 3 gene in the Drosophila melanogaster species subgroup. Analysis of the nucleotide substitution pattern showed that in the coding region synonymous changes occurred more frequently to A or T in contrast to the GC-rich base composition, while in the 3' region the nucleotide substitutions were most likely in equilibrium. These results suggested that the base composition at the third codon position of the H3 gene, i.e., codon usage, has been changing to A or T in the Drosophila melanogaster species subgroup.


Evolutionary change of codon usage for the histone gene family in Drosophila melanogaster and Drosophila hydei.

The nucleotide divergence in the protein-coding region for replication-dependent and replication-independent histone 3 and 4 genes of Drosophila melanogaster and Drosophila hydei occurred mostly at the synonymous site. Therefore, the pattern of codon usage was analyzed in the two species, considering the genomic codon bias, which is proposed for estimating the genomic composition pressure in the protein-coding regions. The results indicated that the codon usage in the histone gene family could be explained mostly by the genomic codon bias. However, biases for Ala and Arg were commonly observed for the histone 3 and histone 4 gene families, and biases for Ser, Leu, and Glu were observed in a gene-specific manner. This suggests that both genomic codon bias and gene- or codon-specific bias are responsible for the nucleotide differentiation in the protein-coding region of the histone genes.


Evolution of the amylase isozymes in the Drosophila melanogaster species subgroup.


Adaptation to a starch environment and regulation of α-amylase in Drosophila.


Divergence of the regulation of α-amylase activity in Drosophila melanogaster, Drosophila funebris and Drosophila saltans.


Genetic analysis of the CYP2D6 gene in patients with Parkinson' disease.


Genetic factors on the second and third chromosomes responsible for the variation of amylase activity and inducibility in Drosophila melanogaster

Using second- or third-chromosome substitution lines of Drosophila melanogaster, the genetic variation of inducibility and amylase specific activities in three media (starch, normal and glucose) were investigated. Genetic factors on both the second and third chromosomes were responsible for the variation in amylase specific activity and inducibility. In glucose medium, the genetic variance of amylase specific activity estimated for the second-chromosome substitution lines was larger than that for the third-chromosome substitution lines; however, for starch medium and inducibility, the variance was larger for the third-chromosome substitution lines. High correlations for the second-chromosome substitution lines and low correlations for the third-chromosome substitution lines were observed for amylase specific activities in different media. These results suggest that the genetic factor(s) responsible for inducibility or amylase activity variation in an induced medium such as starch should be on the third chromosome and those in the non-induced medium such as glucose should be on the second chromosome. The functional roles of the factors on the second and third chromosomes would be the repression and induction of amylase, respectively.


Characterization of the ferredoxin gene transcripts in bovine liver and brain.

1. To study the expression of a ferredoxin gene in extra-adrenocortical tissues, the amounts and structures of the ferredoxin gene transcripts in bovine liver and brain were studied and compared to those of that in adrenocortex. 2. The sizes and amounts of the ferredoxin mRNAs were analyzed by means of Northern blotting, RNA slot blotting and RT-PCR. 3. The nucleotide sequences were determined for 39 ferredoxin cDNA clones from bovine liver. 4. The results indicated that the sizes of the ferredoxin mRNAs in liver and brain were the same as that in adrenocortex, however, their amounts were approx 1/30th and less than the latter, respectively. 5. Although the multiple forms of ferredoxin cDNA, differing in the poly(A) addition sites, were also found in liver, a minor form of ferredoxin cDNA, produced through alternative promoter usage and splicing, could not be detected in liver. 6. The nucleotide sequences of all hepato-ferredoxin cDNA clones obtained were identical to that of a major type of adreno-ferredoxin. 7. These results showed that the expression level of the ferredoxin gene in different tissues was controlled by the amount of mRNA.

The International Journal of Biochemistry


アルコール関連障害とアルコール依存症 アルコール代謝関連酵素ー検査法ー遺伝子構造解析法(Review)


Molecular evolution of the paired gene in Drosophila,Cloning and characterization of the partial paired gene from Drosophila willistoni


Genotype analysis of the CYP2C19 gene in the Japanese population


Novel 2-point linked mutations in the 5'-flanking region of human CYP2C18


Genetic analysis of the cytochrome P-450IIC18(CYP2C18)gene and a novel member of the CYP2C subfamily


Characterstics of Japanese alcoholics with the atypical aldehyde dehydrogenase 2*2. Acomparison of the genotypes of ALDH2,ADH2,ADH3,and cytochrome P-4502E1 between alcoholics and nonalcoholics.


Characteristics of Japanese alcoholics with the atypical aldehyde dehydrogenase 2*2.I.Acomparison of the genotypes of ALDH2,ADH2,ADH3,and cytochrome P-4502E1 between alcoholics and nonalcoholics.

We examined the genotypes of the aldehyde dehydrogenase (ALDH)-2, alcohol dehydrogenase (ADH)-2, ADH3, and P-4502E1 loci of 53 alcoholics and 97 nonalcoholics. All of the subjects fulfilled the DSM-III-R criteria for alcohol dependence. The control group consisted of 97 subjects who were either hospital staff or students. We also compared the frequencies of homozygous ALDH2*1/1 and heterozygous ALDH2*1/2 genotypes in alcoholics. Our study revealed differences in the allelic frequencies of the ALDH2, ADH2, and ADH3 loci between alcoholics and nonalcoholics. For alcoholics with both homozygous ALDH2*1/1 and heterozygous ALDH2*1/2 genotypes, it was found that ADH2 and ADH3 played important rates. Alcoholics with the heterozygous ALDH2*1/2 genotype showed a significantly higher frequency of ADH2*1/1 than ones with the homozygous ALDH2*1/1 genotype. We assume ADH2*1 plays an important role in the development of alcoholism in alcoholics with the heterozygous ALDH2*1/2 genotype.

Alcoholism, Clinical and Experimental Research


CYP2E1 and ALDH2 genotypes and alcohol dependence in Japanese


CYP2E1 genotypes and serum LAP in Japanese Alcoholics


アルコール依存症とALDH2,ADH2,ADH3,P-4502E1との関連について

アルコール研究と薬物依存

アルコール研究と薬物依存


A novel cytochrome P-450IID6(CYPIID6) mutant gene associated with multiple system atrophy

Journal of Neurology, Neurosurgery, and Psychiatry


Relationship between genetic polymorphism of CYP2E1 and ALDH2, and possible susceptibility to alcoholism.


Molecular characterization of a class IV human alcohol dehydrogenase gene(ADH7)


エタノールおよびアルデヒドの代謝速度論的研究ーALDH2およびCYP2E1の遺伝子多型がアルコール代謝に及ぼす影響．

アルコール研究と薬物依存

アルコール研究と薬物依存


抗てんかん薬PHTの代謝と神経系への副作用．

抗てんかん薬PHTの代謝と神経系への副作用．


非活性型アルデヒド脱水素酵素ALDH22を有するアルコール依存症の特徴ー他のアルコール代謝酵素との関連についてー．

アルコール研究と薬物依存

アルコール研究と薬物依存


Correspondence of increased debrisoquine 4 monooxygenase activity with seizure-susceptibility in Mongolian gerbils.

Four drug-metabolizing activities in Wistar rat and Mongolian gerbil liver microsomes were investigated to clarify the biochemical basis of convulsions. No significant associations were observed between the susceptibility to the tonic-clonic seizures in Mongolian gerbils and the drug-metabolizing enzymatic activities of p-nitroanisole O-demethylase, aminopyrine demethylase and benzo[a]pyrene-3-monooxygenase. However, a significant association was observed with the debrisoquine 4-monooxygenase (cytochrome P-450db1-linked monooxygenase system) activity, which is known to metabolize exogenous and endogenous parkinsonism-inducing neurotoxins such as MPTP and TIQ, and to activate carcinogens during the detoxication of xenobiotics. The mean activity of debrisoquine 4-monooxygenase in the seizure-sensitive group (male, 380; female, 350 pmol/h/mg protein) remained significantly higher (about 4-5 times) than that in the seizure-resistant group (male, 90; female, 80 pmol/h/mg protein). More Mongolian gerbils were extensive metabolizers of debrisoquine in the seizure-sensitive group than in the seizure-resistant group. These results suggest a possibility that the cytochrome P-450db1-linked monooxygenase system may activate the potential neurotoxins which are associated with tonic-clonic seizures in the Mongolian gerbil.


In vitro administration of H2 blockers, cimetidine and ranitidine, reduced the contents of the cytochrome P-450IID(CYP2D) subfamily and their activities in rat liver microsomes.


Presence of P-450 D subfamily in DA rat and bovine brains.


シトクロームP-450ⅡD6の多型とパーキンソン病

シトクロームP-450ⅡD6の多型とパーキンソン病


Identification of a microsomal retinoic acid synthase as a microsomal cytochrome P-450-linked monooxygenase system.

1. To characterize an enzyme which metabolizes retinal in liver microsomes, several properties of the enzymatic reaction from retinal to retinoic acid were investigated using rabbit liver microsomes. 2. The maximum pH of the reaction in the liver microsomes was 7.6. 3. The Km and Vmax values for all-trans, 9-cis and 13-cis-retinals were determined. 4. The reaction proceeded in the presence of NADPH and molecular oxygen. 5. The incorporation of one atom of molecular oxygen into retinal was confirmed by using oxygen-18, showing that the reaction comprised monooxygenation, not dehydrogenation. 6. The monooxygenase activity was inhibited by carbon monoxide, phenylisocyanide and anti-NADPH-cytochrome P-450 reductase IgG, but not by anti-cytochrome b5 IgG. 7. The enzymatic activity inhibited by carbon monoxide was photoreversibly restored by light of a wavelength of around 450 nm. 8. The retinal-induced spectra of liver microsomes with three isomeric retinals were type I spectra. 9. The microsomal monooxygenase activity induced by phenobarbital or ethanol were more effective than that by 3-methylcholanthrene, clotrimazole or beta-naphthoflavone. 10. These results showed that the monooxygenase reaction from retinal to retinoic acid in liver microsomes is catalyzed by a cytochrome P-450-linked monooxygenase system.

The International Journal of Biochemistry


Improved methods for genotype determination at the ALDH2 locus using PCR by introducing the MBOII recognition site or conducting secondary PCR.

Biochemistry and Molecular Biology International


Side effects of phenytoin and the drug metabolism by cytochrome P-450 in DA rats.


抗てんかん薬の代謝と神経系への副作用

抗てんかん薬の代謝と神経系への副作用


ALDH2のgenotypeの簡便な決定とアルコール依存症の特徴について．

ALDH2のgenotypeの簡便な決定とアルコール依存症の特徴について．


フラッシング反応と飲酒パターンに及ぼすALDH2とCYP2E1の遺伝的多型

フラッシング反応と飲酒パターンに及ぼすALDH2とCYP2E1の遺伝的多型

アルコール研究と薬物依存

アルコール研究と薬物依存


A novel cytochrome P-450 D6 mutant gene associated with Parkinson's disease.


シトクロームP-450db1における向精神薬代謝と薬物相互作用

シトクロームP-450db1における向精神薬代謝と薬物相互作用


Characterization of the ferredoxin gene transcripts in bovine liver and brain.


Side effects of phenytoin related to the xenobiotics metabolism by cytochrome P-450-linked monooxygenase of liver microsomes in Dark Agouti rats.

The Japanese Journal of Psychiatry and Neurology


シトクロームP-450db1の脳内分布

シトクロームP-450db1の脳内分布


Interleukin 2-induction of cytochrome P450-linked monooxygenase systems of rat liver microsomes.

The effects of interleukin 2 (IL-2), a pivotal cytokine for generating an effective immune response, on rat liver microsomal cytochrome P450-linked monooxygenase systems were investigated by measuring the contents of cytochromes b5 and P450, and the activities of various xenobiotic-metabolizing enzymes [debrisoquine and bufuralol monooxygenases (CYP2D), 7-ethoxycoumarin O-deethylase, benzphetamine N-demethylase, aniline hydroxylase and p-nitroanisole N-demethylase]. The enzymatic activities except for p-nitroanisole N-demethylase and aniline hydroxylase were increased approximately to 1.3-fold of those of untreated liver microsomes following intraperitoneal injection of IL-2 (15 U/rat). However, the amount of immunoreactive b5 protein, and the activities of aniline hydroxylase and p-nitroanisole N-demethylase were not changed by injection of IL-2. To elucidate further the mechanism of the induction of CYP2D by IL-2, quantitative analyses of immunoreactive CYP2D protein and its mRNA were conducted by western blot and slot blot hybridization analyses. The results indicated that IL-2 induced an increase in the amounts of immunoreactive CYP2D protein and its mRNA. These enzymatic activities were thus up-regulated at the mRNA level.


Histamine and l-methyI-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline are competitive inhibitors of debrisoquine 4-monooxygenase in rat liver.


Effects of interleukin 1α on the activities and gene expressions of the cytochrome P450 D subfamily.


シトクロムP-450db1の脳内活性

シトクロムP-450db1の脳内活性


Characterization of the cytochrome P-450 D subfamily in bovine liver:Nucleotide sequences and microheterogeneity.


Debrisoquine 4-monooxygenase and bufuralol 1'- monooxygenase activities in bovine and rabbit tissues.


Analysis of the transcripts of cytochrome P-450 D gene subfamily in bovine adrenal gland.


Expression cloning of a sheep adreno-ferredoxin using the polymerase chain reaction.


Molecular structure of the human alcohol dehydrogenase 3 gene.


Determination of the amino acid sequence of adreno-ferredoxin from sheep adrenocortical mitochondria.

1. An adreno-ferredoxin was purified from sheep adrenocortical mitochondria. 2. Its amino acid composition and amino acid sequence were determined chemically. 3. It was found to be composed of 127 amino acid residues, including two tyrosyl residues. 4. The amino acid sequences of various ferredoxins of the two iron and two sulfur type were compared with respect to amino acid homology.

The International Journal of Biochemistry


Molecular cloning and nucleotide sequences of bovine hepato-ferredoxin cDNA:Identical primary structures of hepato-and adreno-ferredoxins.


Cloning and sequencing of a processed pseudogene derived from a human class alcohol dehydrogenase gene.


The genes for human alchol dehydrogenase β1 and β2 differ by only one nucletide.


Nucleotide variation and divergence in the histone multigene family in Drosophila melanogster.


tRNA derived insertion element in histone gene repeating unit of Drosophila melanogaster.


Molecular structure of the human alcohol dehydrogenase 1 gene.


Naturally occurring variation in the restriction map of the Amy region of Drosophila melanogaster.


Nucleotide sequence of a retrotransposon 297 isolated from Drosophila simulans.


Genetic analysis of natural populations of Drosophila melanogaster in Japan. .Differential regulation of duplicated amylase loci and degree of dominance of amylase activity in different environments.


Genetic analysis of natural populations of Drosophila melanogaster in Japan. .Natural selection on the inducibility,but not on the structural genes,of amylase loci.


Identification of the coding sequence for a reverse transcriptase-like enzyme in a transposable genetic element in Drosophila melanogaster.

The largest group of transposable elements in Drosophila melanogaster, copia-like elements, share some important structural features with and are intimately related in evolution to vertebrate retroviruses. To further clarify the relationship between retroviruses and copia-like transposable elements, we set out to determine the complete nucleotide sequence of the genome of 17.6, which has long terminal repeats homologous in nucleotide sequence to those of avian leukaemia-sarcoma virus. We report here that 17.6 contains three long open reading frames comparable with gag, pol and env genes in retrovirus. At the level of amino acid sequence, the longest open reading frame of 17.6 includes a coding sequence similar to that for reverse transcriptase, suggesting a role for this enzyme in the life cycle of some Drosophila copia-like elements, analogous to the situation in retrovirus.


Genetic analysis of natural populations of Drosophila melanogaster in Japan. .Genetic variability of inducing factors of amylase and fitness.


Cloning of P-elements from P-and Q-strains in natural populations of Drosophila melanogaster in Japan.


二倍体動物(ショウジョウバエ)および半数体植物(蛇苔)における遺伝的変異と進化

二倍体動物(ショウジョウバエ)および半数体植物(蛇苔)における遺伝的変異と進化


Genetic analysis of natural populations of Drosophila melanogaster in Japan. .Protein polymorphism,lethal gene,sterility gene,inversion polymorphism and linkage disequilibrium.


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

Proceedings of the National Academy of Sciences of the United States of America


Genetic variability and selection for inducibility at the amylase locus in Drosophila melanogaster.


ショウジョウバエホメオボックス遺伝子の進化

ショウジョウバエホメオボックス遺伝子の進化