Alcohol Dehydrogenase Locus Research Articles

Recent Selection on a Class I ADH Locus Distinguishes Southwest Asian Populations Including Ashkenazi Jews.

The derived human alcohol dehydrogenase (ADH)1B*48His allele of the ADH1B Arg48His polymorphism (rs1229984) has been identified as one component of an East Asian specific core haplotype that underwent recent positive selection. Our study has been extended to Southwest Asia and additional markers in East Asia. Fst values (Sewall Wright’s fixation index) and long-range haplotype analyses identify a strong signature of selection not only in East Asian but also in Southwest Asian populations. However, except for the ADH2B*48His allele, different core haplotypes occur in Southwest Asia compared to East Asia and the extended haplotypes also differ. Thus, the ADH1B*48His allele, as part of a core haplotype of 10 kb, has undergone recent rapid increases in frequency independently in the two regions after divergence of the respective populations. Emergence of agriculture may be the common factor underlying the evident selection.

Genetic variant repressing ADH1A expression confers susceptibility to esophageal squamous-cell carcinoma

Genome-wide association studies (GWAS) have discovered numerous genetic susceptibility loci including a cluster of alcohol dehydrogenase (ADH) gene family for esophageal squamous-cell carcinoma (ESCC). However, the underlying mechanism has not fully been elucidated. In this study, we integrated the GWAS data, gene-drinking interaction, expression quantitative trait locus (eQTL) analysis and biochemical experiments to clarify the specific mechanism of the polymorphisms in ADH loci. By imputation and eQTL analysis, we identified rs1154402C>G in intron 1 of ADH5 substantially associated with the expression levels of ADH1A. Association analysis showed that the rs1154402[G] allele was significantly associated with ESCC risk in drinkers (OR = 1.44, 95% CI = 1.20-1.73; P = 7.74 × 10-5) but not in nondrinkers (OR = 1.14, 95% CI = 0.93-1.37; P = .220). Furthermore, the ADH5 variant showed a significant interaction with drinking and the genetic variant near ALDH2 encoding the enzyme oxidizing acetaldehyde, a carcinogenic product resulted from alcohol oxidation catalyzed by ADHs. We demonstrated for the first time that rs1154402C>G change might create a silencer, repressing ADH1A transcription via long-range interaction with ADH1A promoter. These results suggest that genetic variant in ADH5 might confer alcohol drinkers susceptible to ESCC by down-regulation of ADH1A, which weakens alcohol catabolism.

Taxonomic status of two morphotypes of Coryphaena hippurus (Perciformes: Coryphaenidae)

ABSTRACT Two Coryphaena hippurus morphotypes (dourado and palombeta) are found along the Brazilian coast and are considered by Rio de Janeiro’s fisherman and fishmongers as two different species. Furthermore, these morphotypes are commercialized under different values and suffer different fishing pressure. Therefore, a definition of their taxonomic status is an important economic and biological matter. In order to investigate this problem, allozyme electrophoresis method was undertaken for seventeen loci on 117 individuals of C. hippurus sampled at Cabo Frio/RJ (Brazil). The data indicate homogeneity between the morphotypes gene pools. Nevertheless, differences were found for genetic variation among dourado and palombeta, especially due to alcohol dehydrogenase locus. Natural selection hypothesis is discussed in explaining these findings.

Evidence of Positive Selection on a Class I ADH Locus

The alcohol dehydrogenase (ADH) family of enzymes catalyzes the reversible oxidation of alcohol to acetaldehyde. Seven ADH genes exist in a segment of ~370 kb on 4q21. Products of the three class I ADH genes that share 95% sequence identity are believed to play the major role in the first step of ethanol metabolism. Because the common belief that selection has operated at the ADH1B*47His allele in East Asian populations lacks direct biological or statistical evidence, we used genomic data to test the hypothesis. Data consisted of 54 single-nucleotide polymorphisms (SNPs) across the ADH clusters in a global sampling of 42 populations. Both the F(st) statistic and the long-range haplotype (LRH) test provided positive evidence of selection in several East Asian populations. The ADH1B Arg47His functional polymorphism has the highest F(st) of the 54 SNPs in the ADH cluster, and it is significantly above the mean F(st) of 382 presumably neutral sites tested on the same 42 population samples. The LRH test that uses cores including that site and extending on both sides also gives significant evidence of positive selection in some East Asian populations for a specific haplotype carrying the ADH1B*47His allele. Interestingly, this haplotype is present at a high frequency in only some East Asian populations, whereas the specific allele also exists in other East Asian populations and in the Near East and Europe but does not show evidence of selection with use of the LRH test. Although the ADH1B*47His allele conveys a well-confirmed protection against alcoholism, that modern phenotypic manifestation does not easily translate into a positive selective force, and the nature of that selective force, in the past and/or currently, remains speculative.

Distinct electrophoretic polymorphism pattern at alcohol dehydrogenase (Adh) locus of Drosophila melanogaster natural populations from Turkey

Small number of Drosophila melanogaster populations from two distinct geographical regions, Central Anatolia and Black Sea, of Turkey were studied. Populations sampled were electrophoresed for a single locus, alcohol dehydrogenase (Adh) to assess population differentiation. Both the magnitude of genetic differentiation levels and the population structure based on hierarchical F-statistics allow populations to be grouped on two genetically divergent area, Central Anatolian and Black Sea. One ecological correlate, average yearly maximum rainfal. Ryear, seems to track this Adh genetic variation pattern. The study also shows that a typical pattern of geographical Adh polymorphism can emerge with a handfull of populations sampled across a relatively small distance.

Distant HNF1 Site as a Master Control for the Human Class I Alcohol Dehydrogenase Gene Expression

Gene duplication and divergence have contributed to the biochemical diversity of the alcohol dehydrogenase (ADH) family. Class I ADH is the major enzyme that catalyzes alcohol to acetaldehyde in the liver. To investigate the mechanism(s) controlling tissue-specific and temporal regulation of the three human class I ADH genes (ADH1A, ADH1B, and ADH1C), we compared genomic sequences for the human and mouse ADH loci and analyzed human ADH gene expression in BAC transgenic mice carrying different lengths of the upstream sequences of the class I ADH. A conserved noncoding sequence, located between the class I and class IV ADH (ADH7) genes, was found to be essential for directing class I ADH gene expression in fetal and adult livers. Within this region, a 275-bp fragment displaying liver-specific DNase I hypersensitivity was bound by HNF1. The HNF1-containing upstream sequence enhanced all three class I ADH promoters in an orientation-dependent manner, and the transcriptional activation depended on binding to the HNF1 site. Deletion of the conserved HNF1 site in the BAC led to the shutdown of human class I ADH gene expression in the transgenic livers, leaving ADH1C gene expression in the stomach unchanged. Moreover, interaction between the upstream element and the class I ADH gene promoters was demonstrated by chromosome conformation capture, suggesting a DNA looping mechanism is involved in gene activation. Taken together, our data indicate that HNF1 binding, at approximately 51 kb upstream, plays a master role in controlling human class I ADH gene expression and may govern alcohol metabolism in the liver.

Genomic organization and functional characterization of the alcohol dehydrogenase locus of Ceratitis capitata (Medfly)

Approximately 30 kb of genomic DNA enclosing the Adh locus from the medfly, Ceratitis capitata have been cloned and about 15 kb has been structurally and functionally characterized. The locus consists of two genes, Adh-1 and Adh-2, separated by an intergenic region, which is polymorphic in size ranging from approximately 6.4 kb to 8.1 kb. Both genes consist of three exons and two introns. The introns are below 200 bp in size, except the 1st intron of Adh-1, which is unexpectedly long, variable in size and contains a deleted mariner-like element (postdoc). The two genes are transcribed in different orientations. The Adh-2 gene shows the typical pattern of transcription seen in the homologous genes of Drosophilidae presenting high levels of expression in the fat body, gut and ovaries. The Adh-1 gene is only expressed in the body muscle tissues of embryos, larvae and adult flies, raising the question of what its biological function may be. A DNA fragment containing bases -102 to -1666 relative to the first base of the initiating ATG of Adh-1 is sufficient to drive the expression of a reporter gene in body muscles of Drosophila melanogaster embryos, larvae and adult flies. The study provides further insights into the evolution of the Adh genes of higher diptera.

An independent non-linear latitudinal cline for the sn-glycerol-3-phosphate (α-Gpdh) polymorphism of Drosophila melanogaster from eastern Australia

Latitudinal variation of the polymorphic sn-glycerol-3-phosphate (alpha-Gpdh) locus in Drosophila melanogaster has been characterized on several continents; however, apparent clinal patterns are potentially confounded by linkage with an inversion, close associations with other genetic markers that vary clinally, and a tandem alpha-Gpdh pseudogene. Here we compare clinal patterns in alpha-Gpdh with those of other linked markers by testing field flies from eastern Australian locations collected in two separate years. The alpha-Gpdh variation exhibited a consistent non-linear cline reflecting an increase in the alpha-GpdhF allele at extreme latitudes. This pattern was not influenced by the In(2L)t inversion wherein this locus is located, nor was it influenced by the presence of the alpha-Gpdh pseudogene, whose presence was ubiquitous and highly variable among populations. The alpha-Gpdh pattern was also independent of a cline in allozyme frequencies at the alcohol dehydrogenase (Adh) locus, and two length polymorphisms in the Adh gene. These results suggest clinal selection at the alpha-Gpdh locus that is partially or wholly unrelated to linear climatic gradients along the eastern coast of Australia.

Activity variation in alcohol dehydrogenase paralogs is associated with adaptation to cactus host use in cactophilicDrosophila

Drosophila mojavensis and Drosophila arizonae are species of cactophilic flies that share a recent duplication of the alcohol dehydrogenase (Adh) locus. One paralog (Adh-2) is expressed in adult tissues and the other (Adh-1) in larvae and ovaries. Enzyme activity measurements of the ADH-2 amino acid polymorphism in D. mojavensis suggest that the Fast allozyme allele has a higher activity on 2-propanol than 1-propanol. The Fast allele was found at highest frequency in populations that utilize hosts with high proportions of 2-propanol, while the Slow allele is most frequent in populations that utilize hosts with high proportions of 1-propanol. This suggests that selection for ADH-2 allozyme alleles with higher activity on the most abundant alcohols is occurring in each D. mojavensis population. In the other paralog, ADH-1, significant differences between D. mojavensis and D. arizonae are associated with a previously shown pattern of adaptive protein evolution in D. mojavensis. Examination of protein sequences showed that a large number of amino acid fixations between the paralogs have occurred in catalytic residues. These changes are potentially responsible for the significant difference in substrate specificity between the paralogs. Both functional and sequence variation within and between paralogs suggests that Adh has played an important role in the adaptation of D. mojavensis and D. arizonae to their cactophilic life.

Variation at isozyme loci in culturedin vitro sugar beet regenerants of gynogenetic origin

Assessment of alcohol dehydrogenase, isocitrate dehydrogenase and 6-phosphogluconate dehydrogenase genes expression has been conducted in culturedin vitro sugar beet vegetative shoots of gynogenetic origin characterized by different level of mixoploidy (x+xx). It was shown that in all 12 originally haploid lines under investigation at alcohol dehydrogenase (ADH1) locus only homozygous phenotypes were observed. At the same time, some of cultured gynogenetic lines demonstrated polymorphism at isocitrate dehydrogenase (IDH1) and 6-phosphogluconate dehydrogenase (PGD1) loci. The conclusion has been done that such a variation in mixoploid tissues of regenerants is associated with spontaneous polyploidization caused by their prolonged culturing underin vitro conditions. Instability of Idh1 and Pgd1 loci, encoding IDH1 and PGD1, together with stability of Adh1 locus, encoding ADH1, testifies that change in ploidy level observed in somatic cells of cultured sugar beet regenerants is unlikely the only factor determining different gene expression duringin vitro culturing.

Tree scanning: a method for using haplotype trees in phenotype/genotype association studies.

We use evolutionary trees of haplotypes to study phenotypic associations by exhaustively examining all possible biallelic partitions of the tree, a technique we call tree scanning. If the first scan detects significant associations, additional rounds of tree scanning are used to partition the tree into three or more allelic classes. Two worked examples are presented. The first is a reanalysis of associations between haplotypes at the Alcohol Dehydrogenase locus in Drosophila melanogaster that was previously analyzed using a nested clade analysis, a more complicated technique for using haplotype trees to detect phenotypic associations. Tree scanning and the nested clade analysis yield the same inferences when permutation testing is used with both approaches. The second example is an analysis of associations between variation in various lipid traits and genetic variation at the Apolipoprotein E (APOE) gene in three human populations. Tree scanning successfully identified phenotypic associations expected from previous analyses. Tree scanning for the most part detected more associations and provided a better biological interpretative framework than single SNP analyses. We also show how prior information can be incorporated into the tree scan by starting with the traditional three electrophoretic alleles at APOE. Tree scanning detected genetically determined phenotypic heterogeneity within all three electrophoretic allelic classes. Overall, tree scanning is a simple, powerful, and flexible method for using haplotype trees to detect phenotype/genotype associations at candidate loci.

Nucleotide polymorphism in the Adh1 locus region of the wild rice Oryza rufipogon.

Nucleotide variation in the alcohol dehydrogenase (Adh1) locus region of the wild rice Oryza rufipogon and its related species was analysed to clarify the maintenance mechanism of DNA variation in Oryza species. The estimated nucleotide diversity in the Adh1 locus region of O. rufipogon was 0.002, which was one of the lowest values detected in nuclear loci of plant species investigated so far. Tests of neutrality detected significantly negative deviation from the neutral mutation model for the coding region, especially for replacement sites. When each of the ADH1 domains was considered, significance was detected only for the catalytic domain 1. These results suggest purifying selection in the Adh1 coding region. In the phylogenetic tree of Oryza species based on Adh1 variation, cultivated rice O. sativa subspp. japonica and indica were included in the cluster of O. rufipogon. The genetic distance of the Adh1 region between O. rufipogon and O. sativa was as low as the nucleotide diversity of O. rufipogon. These results imply that O. rufipogon and O. sativa cannot be classified based on the nucleotide variation of Adh1. No replacement divergence between O. rufipogon and the other three A-genome species (O. glumaepatula, O. barthii and O. meridionalis) were detected, indicating that ADH1 is conserved in the A-genome species. On the other hand, between O. rufipogon and the E-genome species O. australiensis, replacement changes were detected only in the catalytic domain 1. The difference in replacement substitutions between the A- and E-genome species may be related to adaptive changes in the ADH1 domains, reflecting environmental differences where the species encounter anaerobic stress.

Application of Alcohol Dehydrogenase Loci in Testing F<sub>1</sub> Hybridity of Tomato

Expression of alcohol dehydrogenase loci was used to estimate hybridity of Lycopersicon esculentum Mill. in 1012 seeds. The banding patterns were obtained by means of vertical block electrophoresis in polyacrylamide gels. It was established that qualitative variation of locus Adh-2 can be applied to prove hybridity of F1 tomato seeds. This genetic marker is indicative for hybrids with fertile maternal line or with position male sterility line and not only for maternal line with pollen male sterility.

Binge Drinking in Jewish and Non-Jewish White College Students

Background In the United States, religious commitment, as measured by service attendance, has an inverse relationship with alcohol consumption, heavy use, and problem use. This association, however, has not been found consistently in Jewish Americans. The present study examined the relationship between religious variables and binge drinking in Jewish and non-Jewish white college students. In addition, the association among genetic, cultural, and religious variables and binge drinking was examined in the Jewish sample alone. Methods Participants were 132 Jewish and 147 non-Jewish white college students. All participants completed the Time-Line Follow-Back, had blood drawn for genotyping at the alcohol dehydrogenase locus ADH2, and reported their religious affiliation and the number of religious services attended in the past year. Jewish subjects also completed the Jewish Identity Scale. Results As hypothesized, more frequent religious service attendance related to lower rates of binge drinking in non-Jews but was not related to binge drinking in Jews. Within the Jewish sample, individuals who were religiously affiliated had approximately one third the risk of binge drinking as those who were secularly affiliated, but identification with Jewish culture was not related to binge drinking. In the total sample, individuals who possessed a variant alcohol dehydrogenase allele ADH2*2 were approximately half as likely to binge drink as those who did not possess this allele. Conclusions These results are consistent with previous studies that find an inverse relationship between religious service attendance and heavy alcohol use in Christian but not Jewish college students. Findings within the Jewish sample support theories that suggest religious, not just cultural, Jewish affiliation relates to lower levels of alcohol behavior. More research is needed to identify additional factors, including other religious, cultural, genetic, and biological influences, that protect Jewish Americans from heavy drinking.

Binge Drinking in Jewish and Non‐Jewish White College Students

Background In the United States, religious commitment, as measured by service attendance, has an inverse relationship with alcohol consumption, heavy use, and problem use. This association, however, has not been found consistently in Jewish Americans. The present study examined the relationship between religious variables and binge drinking in Jewish and non‐Jewish white college students. In addition, the association among genetic, cultural, and religious variables and binge drinking was examined in the Jewish sample alone.Methods Participants were 132 Jewish and 147 non‐Jewish white college students. All participants completed the Time‐Line Follow‐Back, had blood drawn for genotyping at the alcohol dehydrogenase locus ADH2, and reported their religious affiliation and the number of religious services attended in the past year. Jewish subjects also completed the Jewish Identity Scale.Results As hypothesized, more frequent religious service attendance related to lower rates of binge drinking in non‐Jews but was not related to binge drinking in Jews. Within the Jewish sample, individuals who were religiously affiliated had approximately one third the risk of binge drinking as those who were secularly affiliated, but identification with Jewish culture was not related to binge drinking. In the total sample, individuals who possessed a variant alcohol dehydrogenase allele ADH2*2 were approximately half as likely to binge drink as those who did not possess this allele.Conclusions These results are consistent with previous studies that find an inverse relationship between religious service attendance and heavy alcohol use in Christian but not Jewish college students. Findings within the Jewish sample support theories that suggest religious, not just cultural, Jewish affiliation relates to lower levels of alcohol behavior. More research is needed to identify additional factors, including other religious, cultural, genetic, and biological influences, that protect Jewish Americans from heavy drinking.

The influence of linkage and inbreeding on patterns of nucleotide sequence diversity at duplicate alcohol dehydrogenase loci in wild barley (Hordeum vulgare ssp. spontaneum).

Patterns of nucleotide sequence diversity are analyzed for three duplicate alcohol dehydrogenase loci (adh1-adh3) within a species-wide sample of 25 accessions of wild barley (Hordeum vulgare ssp. spontaneum). The adh1 and adh2 loci are tightly linked (recombination fraction <0.01) while the adh3 locus is inherited independently. Wild barley is predominantly self-fertilizing (approximately 98%), and as a consequence, effective recombination is restricted by the extreme reduction in heterozygosity. Large reductions in effective recombination, in turn, widen the conditions for linkage to influence nucleotide sequence diversity through the action of selective sweeps or background selection. These considerations would appear to predict (1) homogeneity in patterns of nucleotide sequence diversity, especially between closely linked loci, and (2) extensive linkage disequilibrium relative to random-mating species. In contrast to these expectations, the wild barley data reveal heterogeneity in patterns of nucleotide sequence diversity and levels of linkage disequilibrium that are indistinguishable from those observed at adh1 in maize, an outbreeding grass species.

Minisatellite instability at the Adh locus reveals somatic polymorphism in amphioxus.

Amphioxus (subphylum Cephalochordata) is the closest living relative to vertebrates and widely used for phylogenetic analyses of vertebrate gene evolution. Amphioxus genes are highly polymorphic, but the origin and nature of this variability is unknown. We have analyzed the alcohol dehydrogenase locus (Adh3) in two amphioxus species (Branchiostoma lanceolatum and Branchiostoma floridae) and found that genetic variation is related to repetitive DNA sequences, mainly minisatellites. Small pool-PCR assays indicated that allelic variants are generated by minisatellite instability. We conclude that the generation of new forms was not preferentially linked to germline processes but rather to somatic events leading to mosaic adult animals. Furthermore, most Adh minisatellites belong to a novel class, which we have named mirages. Their distinctive feature is that the repeat subunit spans the exon-intron boundaries and generates potential duplications of the splice sites. However, splicing may not be compromised as no aberrant mRNA variants were detected.

Genetic variability among Turkish pop, flint and dent corn (Zea mays L. spp. mays) races: Morphological and agronomic traits

32 Turkish corn accessions representing different climatic, geographic and topographic areas in Turkey were identified. Genetic variation and its pattern in three corn races (pop, flint and dent) included 32 accessions were investigated using 25 morphological and agronomic traits. Vegetative, ear and kernel related traits were significantly varied both among races and accessions within races. Variance component due to races and accessions within race were statistically significant, but the variance components due to accessions were generally made up large component of total variance and ranged from 17.17% in kernel length to 82.75% in tassel length. There were also strong and mostly positive correlations between studied agronomic traits as well as between agronomic traits and isoenzyme loci. These correlations were explained by the traditional farming practices and enzyme specificity with flint accessions since the enzyme systems such as Alcohol dehydrogenase (locus 1), Malate dehydrogenase enzyme (loci 2–3) and 6-phospho-gluconate dehydrogenase (locus-1) are generally considered to be the characteristics of flint corn race. Canonical discriminant function analysis with twenty-five morphological and agronomic traits revealed that first two canonical discriminant variables explained 68% of total variation among accessions. Two Canonical variables, in which kernel related traits were heavily weighted, correctly classify the three corn races – indicating that Turkish corn accessions maintained their racial characteristics with agronomic and morphological traits.

Polysubstance Abuse–Vulnerability Genes: Genome Scans for Association, Using 1,004 Subjects and 1,494 Single-Nucleotide Polymorphisms

Strong genetic contributions to drug abuse vulnerability are well documented, but few chromosomal locations for human drug-abuse vulnerability alleles have been confirmed. We now identify chromosomal markers whose alleles distinguish drug abusers from control individuals in each of two samples, on the basis of pooled-sample microarray and association analyses. Reproducibly positive chromosomal regions defined by these markers in conjunction with previous results were especially unlikely to have been identified by chance. Positive markers identify the alcohol dehydrogenase (ADH) locus, flank the brain-derived neurotropic factor (BDNF) locus, and mark seven other regions previously linked to vulnerability to nicotine or alcohol abuse. These data support polygenic contributions of common allelic variants to polysubstance abuse vulnerability.

Characterization of Two Alcohol Dehydrogenase (Adh) Loci from the Olive Fruit Fly, Bactrocera (Dacus) oleae and Implications for Adh Duplication in Dipteran Insects

We report the cloning and structural characterization of two Adh loci of the olive fruit fly, Bactrocera oleae. Each of the two genes, named Adh1 and Adh2, consists of three exons and two introns for a total length of 1981 and 988 nucleotides, respectively. Their deduced amino acid sequences of 257 and 258 residues exhibit a 77% identity and display the characteristics of the insect ADH enzymes, which belong to the short-chain dehydrogenases/reductases family. The Adh genes of B. oleae are compared to the two genes of the Mediterranean fly, Ceratitis capitata, the only other species of the Tephritidae family in which the Adh genes have been studied. On the basis of amino acid divergence the four genes form two clusters each containing one gene from each species, as expected if there was one duplication event before speciation. On the basis of nucleotide sequence the four sequences form two clusters each containing the two sequences from the same species, as expected if there was a separate duplication event in each species. To help decide between the two alternatives, we compared at both the amino acid and DNA level the Adh genes of five Drosophila species that are known to carry two such genes and observed that, with only one exception at the amino acid level, conspecific loci cluster together. We conclude that the information we have at present does not allow a firm choice between the hypothesis of a single duplication event that occurred before the split of Bactrocera and Ceratitis from their common ancestor and the hypothesis of two independent duplication events, one in each of the two genera.