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. 1993 Nov 1;90(21):10350–10354. doi: 10.1073/pnas.90.21.10350

An ethoxyquin-inducible aldehyde reductase from rat liver that metabolizes aflatoxin B1 defines a subfamily of aldo-keto reductases.

E M Ellis 1, D J Judah 1, G E Neal 1, J D Hayes 1
PMCID: PMC47772  PMID: 8234296

Abstract

Protection of liver against the toxic and carcinogenic effects of aflatoxin B1 (AFB1) can be achieved through the induction of detoxification enzymes by chemoprotectors such as the phenolic antioxidant ethoxyquin. We have cloned and sequenced a cDNA encoding an aldehyde reductase (AFB1-AR), which is expressed in rat liver in response to dietary ethoxyquin. Expression of the cDNA in Escherichia coli and purification of the recombinant enzyme reveals that the protein exhibits aldehyde reductase activity and is capable of converting the protein-binding dialdehyde form of AFB1-dihydrodiol to the nonbinding dialcohol metabolite. We show that the mRNA encoding this enzyme is markedly elevated in the liver of rats fed an ethoxyquin-containing diet, correlating with acquisition of resistance to AFB1. AFB1-AR represents the only carcinogen-metabolizing aldehyde reductase identified to date that is induced by a chemoprotector. Alignment of the amino acid sequence of AFB1-AR with other known and putative aldehyde reductases shows that it defines a subfamily within the aldo-keto reductase superfamily.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adams M. D., Dubnick M., Kerlavage A. R., Moreno R., Kelley J. M., Utterback T. R., Nagle J. W., Fields C., Venter J. C. Sequence identification of 2,375 human brain genes. Nature. 1992 Feb 13;355(6361):632–634. doi: 10.1038/355632a0. [DOI] [PubMed] [Google Scholar]
  2. Benson A. M., Batzinger R. P., Ou S. Y., Bueding E., Cha Y. N., Talalay P. Elevation of hepatic glutathione S-transferase activities and protection against mutagenic metabolites of benzo(a)pyrene by dietary antioxidants. Cancer Res. 1978 Dec;38(12):4486–4495. [PubMed] [Google Scholar]
  3. Bock K. W., Kahl R., Lilienblum W. Induction of rat hepatic UDP-glucuronosyltransferases by dietary ethoxyquin. Naunyn Schmiedebergs Arch Pharmacol. 1980 Jan;310(3):249–252. doi: 10.1007/BF00499919. [DOI] [PubMed] [Google Scholar]
  4. Bohren K. M., Bullock B., Wermuth B., Gabbay K. H. The aldo-keto reductase superfamily. cDNAs and deduced amino acid sequences of human aldehyde and aldose reductases. J Biol Chem. 1989 Jun 5;264(16):9547–9551. [PubMed] [Google Scholar]
  5. Buller A. L., Sharp R. B., Penning T. M. Characterization of dihydrodiol dehydrogenase in rat H-4IIe hepatoma cells. Cancer Res. 1989 Dec 15;49(24 Pt 1):6976–6980. [PubMed] [Google Scholar]
  6. Burbach K. M., Poland A., Bradfield C. A. Cloning of the Ah-receptor cDNA reveals a distinctive ligand-activated transcription factor. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):8185–8189. doi: 10.1073/pnas.89.17.8185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cabral J. R., Neal G. E. The inhibitory effects of ethoxyquin on the carcinogenic action of aflatoxin B1 in rats. Cancer Lett. 1983 Jun;19(2):125–132. doi: 10.1016/0304-3835(83)90146-5. [DOI] [PubMed] [Google Scholar]
  8. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  9. Ciaccio P. J., Stuart J. E., Tew K. D. Overproduction of a 37.5-kDa cytosolic protein structurally related to prostaglandin F synthase in ethacrynic acid-resistant human colon cells. Mol Pharmacol. 1993 Jun;43(6):845–853. [PubMed] [Google Scholar]
  10. Curran T., Franza B. R., Jr Fos and Jun: the AP-1 connection. Cell. 1988 Nov 4;55(3):395–397. doi: 10.1016/0092-8674(88)90024-4. [DOI] [PubMed] [Google Scholar]
  11. Dalrymple B. P., Peters J. M., Vuocolo T. Characterisation of genes encoding two novel members of the aldo-keto reductase superfamily. Biochem Int. 1992 Dec;28(4):651–657. [PubMed] [Google Scholar]
  12. Denison M. S., Fisher J. M., Whitlock J. P., Jr The DNA recognition site for the dioxin-Ah receptor complex. Nucleotide sequence and functional analysis. J Biol Chem. 1988 Nov 25;263(33):17221–17224. [PubMed] [Google Scholar]
  13. Ding V. D., Pickett C. B. Transcriptional regulation of rat liver glutathione S-transferase genes by phenobarbital and 3-methylcholanthrene. Arch Biochem Biophys. 1985 Aug 1;240(2):553–559. doi: 10.1016/0003-9861(85)90062-1. [DOI] [PubMed] [Google Scholar]
  14. Fitzgibbon J. E., Braymer H. D. Cloning of a gene from Pseudomonas sp. strain PG2982 conferring increased glyphosate resistance. Appl Environ Microbiol. 1990 Nov;56(11):3382–3388. doi: 10.1128/aem.56.11.3382-3388.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Friling R. S., Bensimon A., Tichauer Y., Daniel V. Xenobiotic-inducible expression of murine glutathione S-transferase Ya subunit gene is controlled by an electrophile-responsive element. Proc Natl Acad Sci U S A. 1990 Aug;87(16):6258–6262. doi: 10.1073/pnas.87.16.6258. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fujisawa-Sehara A., Sogawa K., Yamane M., Fujii-Kuriyama Y. Characterization of xenobiotic responsive elements upstream from the drug-metabolizing cytochrome P-450c gene: a similarity to glucocorticoid regulatory elements. Nucleic Acids Res. 1987 May 26;15(10):4179–4191. doi: 10.1093/nar/15.10.4179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hayes J. D., Judah D. J., McLellan L. I., Kerr L. A., Peacock S. D., Neal G. E. Ethoxyquin-induced resistance to aflatoxin B1 in the rat is associated with the expression of a novel alpha-class glutathione S-transferase subunit, Yc2, which possesses high catalytic activity for aflatoxin B1-8,9-epoxide. Biochem J. 1991 Oct 15;279(Pt 2):385–398. doi: 10.1042/bj2790385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hayes J. D., Judah D. J., McLellan L. I., Neal G. E. Contribution of the glutathione S-transferases to the mechanisms of resistance to aflatoxin B1. Pharmacol Ther. 1991;50(3):443–472. doi: 10.1016/0163-7258(91)90053-o. [DOI] [PubMed] [Google Scholar]
  19. Hayes J. D., Judah D. J., Neal G. E. Resistance to aflatoxin B1 is associated with the expression of a novel aldo-keto reductase which has catalytic activity towards a cytotoxic aldehyde-containing metabolite of the toxin. Cancer Res. 1993 Sep 1;53(17):3887–3894. [PubMed] [Google Scholar]
  20. Imaida K., Fukushima S., Shirai T., Ohtani M., Nakanishi K., Ito N. Promoting activities of butylated hydroxyanisole and butylated hydroxytoluene on 2-stage urinary bladder carcinogenesis and inhibition of gamma-glutamyl transpeptidase-positive foci development in the liver of rats. Carcinogenesis. 1983;4(7):895–899. doi: 10.1093/carcin/4.7.895. [DOI] [PubMed] [Google Scholar]
  21. Inoue S., Sharma R. C., Schimke R. T., Simoni R. D. Cellular detoxification of tripeptidyl aldehydes by an aldo-keto reductase. J Biol Chem. 1993 Mar 15;268(8):5894–5898. [PubMed] [Google Scholar]
  22. Judah D. J., Hayes J. D., Yang J. C., Lian L. Y., Roberts G. C., Farmer P. B., Lamb J. H., Neal G. E. A novel aldehyde reductase with activity towards a metabolite of aflatoxin B1 is expressed in rat liver during carcinogenesis and following the administration of an anti-oxidant. Biochem J. 1993 May 15;292(Pt 1):13–18. doi: 10.1042/bj2920013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kahl R. Enhancement of epoxide hydratase activity in rat lung, kidney and liver by dietary antioxidants. Cancer Lett. 1980 Feb;8(4):323–328. doi: 10.1016/0304-3835(80)90148-2. [DOI] [PubMed] [Google Scholar]
  24. Kensler T. W., Egner P. A., Dolan P. M., Groopman J. D., Roebuck B. D. Mechanism of protection against aflatoxin tumorigenicity in rats fed 5-(2-pyrazinyl)-4-methyl-1,2-dithiol-3-thione (oltipraz) and related 1,2-dithiol-3-thiones and 1,2-dithiol-3-ones. Cancer Res. 1987 Aug 15;47(16):4271–4277. [PubMed] [Google Scholar]
  25. Kim H. L., Anderson A. C., Herrig B. W., Jones L. P., Calhoun M. C. Protective effects of antioxidants on bitterweed (Hymenoxys odorata DC) toxicity in sheep. Am J Vet Res. 1982 Nov;43(11):1945–1950. [PubMed] [Google Scholar]
  26. Miranda C. L., Carpenter H. M., Cheeke P. R., Buhler D. R. Effect of ethoxyquin on the toxicity of the pyrrolizidine alkaloid monocrotaline and on hepatic drug metabolism in mice. Chem Biol Interact. 1981 Oct;37(1-2):95–107. doi: 10.1016/0009-2797(81)90168-x. [DOI] [PubMed] [Google Scholar]
  27. Molowa D. T., Wrighton S. A., Blanke R. V., Guzelian P. S. Characterization of a unique aldo-keto reductase responsible for the reduction of chlordecone in the liver of the gerbil and man. J Toxicol Environ Health. 1986;17(4):375–384. doi: 10.1080/15287398609530832. [DOI] [PubMed] [Google Scholar]
  28. Oliver S. G., van der Aart Q. J., Agostoni-Carbone M. L., Aigle M., Alberghina L., Alexandraki D., Antoine G., Anwar R., Ballesta J. P., Benit P. The complete DNA sequence of yeast chromosome III. Nature. 1992 May 7;357(6373):38–46. doi: 10.1038/357038a0. [DOI] [PubMed] [Google Scholar]
  29. Parke D. V., Rahim A., Walker R. Reversibility of hepatic changes caused by ethoxyquin. Biochem Pharmacol. 1974 Jul 1;23(13):1871–1876. doi: 10.1016/0006-2952(74)90196-8. [DOI] [PubMed] [Google Scholar]
  30. Pawlowski J. E., Huizinga M., Penning T. M. Cloning and sequencing of the cDNA for rat liver 3 alpha-hydroxysteroid/dihydrodiol dehydrogenase. J Biol Chem. 1991 May 15;266(14):8820–8825. [PubMed] [Google Scholar]
  31. Prestera T., Holtzclaw W. D., Zhang Y., Talalay P. Chemical and molecular regulation of enzymes that detoxify carcinogens. Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):2965–2969. doi: 10.1073/pnas.90.7.2965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Prochaska H. J., De Long M. J., Talalay P. On the mechanisms of induction of cancer-protective enzymes: a unifying proposal. Proc Natl Acad Sci U S A. 1985 Dec;82(23):8232–8236. doi: 10.1073/pnas.82.23.8232. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rao M. S., Lalwani N. D., Watanabe T. K., Reddy J. K. Inhibitory effect of antioxidants ethoxyquin and 2(3)-tert-butyl-4-hydroxyanisole on hepatic tumorigenesis in rats fed ciprofibrate, a peroxisome proliferator. Cancer Res. 1984 Mar;44(3):1072–1076. [PubMed] [Google Scholar]
  34. Rondeau J. M., Tête-Favier F., Podjarny A., Reymann J. M., Barth P., Biellmann J. F., Moras D. Novel NADPH-binding domain revealed by the crystal structure of aldose reductase. Nature. 1992 Jan 30;355(6359):469–472. doi: 10.1038/355469a0. [DOI] [PubMed] [Google Scholar]
  35. Rushmore T. H., Pickett C. B. Transcriptional regulation of the rat glutathione S-transferase Ya subunit gene. Characterization of a xenobiotic-responsive element controlling inducible expression by phenolic antioxidants. J Biol Chem. 1990 Aug 25;265(24):14648–14653. [PubMed] [Google Scholar]
  36. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sparnins V. L., Chuan J., Wattenberg L. W. Enhancement of glutathione S-transferase activity of the esophagus by phenols, lactones, and benzyl isothiocyanate. Cancer Res. 1982 Apr;42(4):1205–1207. [PubMed] [Google Scholar]
  38. Talalay P., Benson A. M. Elevation of quinone reductase activity by anticarcinogenic antioxidants. Adv Enzyme Regul. 1982;20:287–300. doi: 10.1016/0065-2571(82)90021-8. [DOI] [PubMed] [Google Scholar]
  39. Watanabe K., Fujii Y., Nakayama K., Ohkubo H., Kuramitsu S., Kagamiyama H., Nakanishi S., Hayaishi O. Structural similarity of bovine lung prostaglandin F synthase to lens epsilon-crystallin of the European common frog. Proc Natl Acad Sci U S A. 1988 Jan;85(1):11–15. doi: 10.1073/pnas.85.1.11. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wattenberg L. W. Chemoprevention of cancer. Cancer Res. 1985 Jan;45(1):1–8. [PubMed] [Google Scholar]
  41. Wattenberg L. W. Inhibition of carcinogenic and toxic effects of polycyclic hydrocarbons by phenolic antioxidants and ethoxyquin. J Natl Cancer Inst. 1972 May;48(5):1425–1430. [PubMed] [Google Scholar]
  42. Wilson D. K., Bohren K. M., Gabbay K. H., Quiocho F. A. An unlikely sugar substrate site in the 1.65 A structure of the human aldose reductase holoenzyme implicated in diabetic complications. Science. 1992 Jul 3;257(5066):81–84. doi: 10.1126/science.1621098. [DOI] [PubMed] [Google Scholar]
  43. Winters C. J., Molowa D. T., Guzelian P. S. Isolation and characterization of cloned cDNAs encoding human liver chlordecone reductase. Biochemistry. 1990 Jan 30;29(4):1080–1087. doi: 10.1021/bi00456a034. [DOI] [PubMed] [Google Scholar]

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