The Functional Consequences of Polymorphisms in the Human PON1 Gene

  • C.E. Furlong
  • R.J Richter
  • W.-F. Li
  • V.H. Brophy
  • C. Carlson
  • M. Rieder
  • D. Nickerson
  • L.G. Costa
  • J. Ranchalis
  • A.J. Lusis
  • D.M. Shih
  • A. Tward
  • G.P. Jarvik

Abstract

Early research on population distributions of plasma PON1 paraoxonase activity indicated a polymorphic distribution with high, intermediate and low metabolizers. Cloning and characterization of the cDNA encoding human PON1 and follow-on experiments demonstrated that the molecular basis of the activity polymorphism (PM) was a Q192R PM with PON1R192 specifying high paraoxonase activity. Further research demonstrated that the PON1192 polymorphism had little effect on the catalytic efficiencies of hydrolysis of phenylacetate and diazoxon (DZO), but did affect the efficiencies of hydrolysis of chlorpyrifos oxon (CPO), soman and sarin, with PON1R192 having a higher efficiency of CPO hydrolysis and PON1Q192 having higher rates of hydrolysis of soman and sarin. Plots of rates of DZO hydrolysis (at a salt concentration that differentially inhibited PON1R192) vs. paraoxon hydrolysis clearly separated the three PON1192 phenotypes (QQ, QR, RR) and also showed a wide range of activity among individuals with the same PON1192 genotype. The term PON1 status was introduced to include both PON1192 functional genotype and plasma PON1 level,both important in determining risk for either exposure to specific organophosphorus compounds (OPs) or disease. Characterization of 5 promoter-region polymorphisms by several groups indicated that an Sp1 binding site was responsible for significant(~30%) variation in plasma PON1 levels. Re-sequencing of the PON1 genes of 47 individuals (24 African-American/23 European) revealed an additional 180 polymorphisms in 27 kb of the PON1 genomic DNA including 8 more 5' regulatory region PMs, 1 coding region polymorphism (W194X), 162 additional intronic PMs and 9 additional 3' UTR PMs. The generation of PON1 null mice and “PON1 humanized mice” expressing either tgHuPON1R192 or tgHuPON1Q192 at the same levels on the PON1−/− background allowed for a functional analysis of the Q192R PM under physiological conditions. Toxicology experiments with the PON1 humanized mice and the PON1 null mice injected with purified human PON1192 alloforms clearly demonstrated that the catalytic efficiency of substrate hydrolysis is important in determining whether PON1 is able to protect against a given OP exposure. HuPON1R192 protects well against CPO and DZO exposure, but HuPON1Q192 does not protect well against CPO exposure and neither protects against PO exposure. Studies on PON1 status and carotid artery disease show that low PON1 levels are a risk factor. The effects of PON1192 alloforms on rates of hydrolysis of quorum sensing factors are not yet known. Taken together, these data along with those of the leading researchers in the PON1 field indicate that it is important to measure PON1 levels/activities in any epidemiological study. SNP analysis alone is inadequate for epidemiological studies, due to the wide variability of PON1 levels within the three PON1192 genotypes Q/Q, Q/R R/R). Even the most comprehensive PON1 SNP analyses are unable to accurately predict PON1 levels. PON1 activity or level accurately predicts CHD risk, while genotype does not

Keywords

paraoxonase PON1 organophosphate organophosphorus compounds (OPs) chlorpyrifos chlorpyrifos oxon diazinon diazoxon arylesterase regulation of gene expression nerve agents single nucleotide polymorphisms (SNPs) PON1 expression Sp1 transcription factor quorum sensing quorum sensing factor carotid artery disease (CAAD) coronary heart disease (CHD) 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Reference

  1. Adkins, S., Gan, K.N., Mody, M., La Du, B.N., 1993, Molecular basis for the polymorphic forms of human serum paraoxonase/arylesterase: Glutamine or arginine at position 191 for the respective A or B allozymes. Am. J Hum. Gene. 52:598–608Google Scholar
  2. Blatter Garin, M.-C., James, R.W., Dussoix, P., et al., 1997, Paraoxonase polymorphism Met-Leu54 is associated with modified serum concentrations of the enzyme. A possible link between the paraoxonase gene and increased risk of cardiovascular disease in diabetes. J. Clin. Invest. 99:62–66CrossRefGoogle Scholar
  3. Brophy, V.H., Hastings, M.D., Clendenning, J.B., Richter, R.J., Jarvik, G.P., Furlong, C.E., 2001a, Polymorphisms in the human paraoxonase (PON1) promoter. Pharmacogenetics 11:77–84CrossRefGoogle Scholar
  4. Brophy, V.H., Jampsa, R.L., Clendenning, J.B., Jarvik, G.P., Furlong, C.E., (2001b), Promoter polymorphisms affect paraoxonase (PON1) expression. Am. J. Hum. Genet. 68:1428–1436CrossRefGoogle Scholar
  5. Chambers, J.E., Ma, R., Boone, J.S., Chambers, H.W., 1994, Role of detoxication pathways in acute toxicity of phosphorothioate insecticides in the rat. Life Sci. 54:1357–1364CrossRefPubMedGoogle Scholar
  6. Costa, L.G., Vitalone, A., Cole, T.B., Furlong, C.E., 2005, Modulation of paraoxonase (PON1) activity. Biochem Pharmacol., 15,69(4):541–550CrossRefGoogle Scholar
  7. Deakin, S.P., James, R.W., 2004, Genetic and environmental factors modulating serum concentrations and activities of the antioxidant enzyme paraoxonase-1. Clin Sci (Lond). 107(5):435–447CrossRefGoogle Scholar
  8. Deakin, S., Leviev, I., Brulhart-Meynet, M.C., James, R.W., 2003, Paraoxonase-1 promoter haplotypes and serum paraoxonase: a predominant role for polymorphic position - 107, implicating the Sp1 transcription factor. Biochem, J. 372(Pt 2):643–649CrossRefGoogle Scholar
  9. Davies, H., Richter, R.J., Keifer, M., Broomfield, C., Sowalla, J., Furlong, C.E., 1996, The effect of the human serum paraoxonase polymorphism is reversed with diazoxon, soman and sarin. Nature Genet. 14:334–336CrossRefPubMedGoogle Scholar
  10. Furlong, C., Holland, N., Richter, R., Bradman, A., Ho, A., Eskenazi, B., 2006, PON1 status of farmworker mothers and children as a predictor of organophosphate sensitivity. Pharmacogenetics and Genomics. 16:183–190PubMedGoogle Scholar
  11. Hantusch, B., Kalt, R., Krieger, S., Puri, C., Kerjaschki, D., 2007, Sp1/Sp3 and DNA-methylation contribute to basal transcriptional activation of human podoplanin in MG63 versus Saos-2 osteoblastic cells. BMC Mol Biol., 8:20CrossRefPubMedGoogle Scholar
  12. Harel, M., Aharoni, A., Gaidukov, L., Brumshtein, B., Khersonsky, O., Meged, R., Dvir, H., Ravelli, R.B.G., McCarthy, A., Toker, L., Silman, I., Sussman, J.L., Tawfik, D.S., 2004, Structure and evolution of the serum paraoxonase family of detoxifying and anti-atherosclerotic enzymes. Nature Structural & Molecular Biology 11:412–419CrossRefGoogle Scholar
  13. Hassett, C., Richter, R.J., Humbert, R., Chapline, C., Crabb, J.W., Omiecinski, C.J., Furlong, C.E., 1991, Characterization of cDNA clones encoding rabbit and human serum paraoxonase: the mature protein retains its signal sequence. Biochemistry 30:10141–10149CrossRefPubMedGoogle Scholar
  14. Huff, R., Abou-Donia, M.B., 1995, In vitro effect of chlorpyrifos oxon on muscarinic receptors and adenylate cyclase. Neurotoxicology 16:281–290PubMedGoogle Scholar
  15. Humbert, R., Adler, D.A., Disteche, C.M., Hassett, C., Omiecinski, C.J., Furlong, C.E., 1993, The molecular basis of the human serum paraoxonase activity polymorphism. Nat. Genet., 3:73–76CrossRefPubMedGoogle Scholar
  16. Jarvik, G.P., Jampsa, R., Richter, R.J., Carlson, C., Rieder, M., Nickerson, D., Furlong, C.E., 2003, Novel Paraoxonase (PON1) nonsense and missense mutations predicted by functional genomic assay of PON1 status. Pharmacogenetics 13:291–295CrossRefPubMedGoogle Scholar
  17. Jokanovic, M., 2001, Biotransformation of organophosphorus compounds. Toxicology 166:139–160CrossRefPubMedGoogle Scholar
  18. La Du, BN., 2003, Future studies of low-activity PON1 phenotype subjects may reveal how PON1 protects against cardiovascular disease. Arterioscler. Thromb. Vasc. Biol., 23(8):1317–1318CrossRefGoogle Scholar
  19. Lawlor, D.A., Day, I.N., Gaunt, T.R., Hinks, L.J., Briggs, P.J., Kiessling, M., Timpson, N., Smith, G.D., Ebrahim, S., 2004, The association of the PON1 Q192R polymorphism with coronary heart disease: findings from the British Women’s Heart and Health cohort study and a meta-analysis. BMC Genet., 5:17CrossRefPubMedGoogle Scholar
  20. Leviev, I., Deakin, S., James, R.W., 2001, Decreased stability of the M54 isoform of paraoxonase as a contributory factor to variations in human serum paraoxonase concentrations. J. Lipid Res. 42:528–535PubMedGoogle Scholar
  21. Leviev, I., James, R.W., 2000, Promoter polymorphisms of the human paraoxonase PON1 gene and serum paraoxonase activities and concentrations. Arterioscler. Thromb. Vasc. Biol. 20:516–552PubMedGoogle Scholar
  22. Leviev, I., Negro, F., James, R.W., 1997, Two alleles of the human paraoxonase gene produce different amounts of mRNA: an explanation for differences in serum concentrations of paraoxonase associated with the (Leu-Met54) polymorphism. Arterioscler. Thromb. Vasc. Biol. 17:3935–3939Google Scholar
  23. Li, H.-L., Liu, D.-P., Liang, C.-H., 2003, Paraoxonase gene polymprphisms, oxidative stress and diseases. J. Mol. Med, 81:766–779CrossRefPubMedGoogle Scholar
  24. Li, W.-F., Costa, L.G., Richter, R.J., Hagen, T., Shih, D.M., Tward, A., Lusis A.J., Furlong, C.E., 2000, Catalytic efficiency determines the in vivo efficacy of PON1 for detoxifying organophosphates. Pharmacogenetics, 10:767–780CrossRefPubMedGoogle Scholar
  25. Ling, J.Q., Hoffman, A.R., 2007, Epigenetics of long-range chromatin interactions. Pediatr Res. 2007 Mar 15; [Epub ahead of print]Google Scholar
  26. Mackness, B., Davie, G.K., Turkie, W., Lee, E., Roberts, D.H., Hill, E., Roberts, C., Durrington, P.N., Mackness, M.I., 2001, Paraoxonase status in coronary heart disease: are activity and concentration more important than genotype?. Arterioscler. Thromb. Vasc. Biol., 21(9):1451–1457CrossRefPubMedGoogle Scholar
  27. Mackness, B., Durrington, P.N., Mackness, M., 2002, PON1 in other diseases. In Paraoxonase (PON1) in Health and Disease: Basic and Clinical Aspects (L.G. Costa and C.E. Furlong, eds.), Kluwer Academic Publishers, Boston, USA, pp. 185–195Google Scholar
  28. Mackness, M., Arrol, S., Durrington, P.N., 1991, Paraoxonase prevents accumulation of lipoperoxides in low-density lipoprotein. FEBS Lett., 286:152–154CrossRefPubMedGoogle Scholar
  29. Pond, A.L., Chambers, H.W., Chambers, J.E., 1995, Organophosphate detoxication potential of various rat tissues via A-esterase and aliesterase activities. Toxicol. Lett. 70:245–252CrossRefGoogle Scholar
  30. Roest, M., van Himbergen, T.M., Barendrecht, A.B., Peeters, P.H., van der Schouw, Y.T., Voorbij, H.A., 2007, Genetic and environmental determinants of the PON-1 phenotype. Eur, J Clin, Invest., 37(3):187–196Google Scholar
  31. Saikawa, Y., Price, K., Hance, K.W., Chen, T.Y., Elwood, P.C., 1995, Structural and functional analysis of the human KB cell folate receptor gene P4 promoter: cooperation of 3 clustered Sp1-binding sites with the initiator region for basal promoter activity. Biochemistry 34:995–9961CrossRefGoogle Scholar
  32. Shih, D.M., Gu, L., Xia, Y.-R., Navab, M., Li, W.-F., Hama, S., Castellani, L.W., Furlong, C.E., Costa, L.G., Fogelman, A.M., Lusis, A.J., 1998, Mice lacking serum paraoxonase are susceptible to organophosphate toxicity and atherosclerosis. Nature 394:284–287.CrossRefPubMedGoogle Scholar
  33. Suehiro, T., Nakamura, T., Inoue, M., Shiinoki, T., Ikeda, Y., Kumon, Y., Shindo, M., et al., 2000, A polymorphism upstream from the human paraoxonase (PON1) gene and its association with PON1 expression. Atherosclerosis 150:295–298CrossRefPubMedGoogle Scholar
  34. Usmani, K.A., Cho, T.M., Rose, R.L., Hodgson, E., 2006, Inhibition of the human liver microsomal and human cytochrome P450 1A2 and 3A4 metabolism of estradiol by deployment-related and other chemicals. Drug Metab. Dispos., 34(9):1606–1614CrossRefPubMedGoogle Scholar
  35. Usmani, K.A., Rose, R.L., Hodgson, E., 2003, Inhibition and activation of the human liver microsomal and human cytochrome P450 3A4 metabolism of testosterone by deployment-related chemicals. Drug Metab. Dispos., 31(4):384–91.CrossRefPubMedGoogle Scholar
  36. Wheeler, J.G., Keavney, B.D., Watkins, H., Collins, R., Danesh, J., 2004, Four paraoxonase gene polymorphisms in 11212 cases of coronary heart disease and 12786 controls: meta-analysis of 43 studies. Lancet, 363(9410):689–695CrossRefPubMedGoogle Scholar
  37. Winnier, D.A, Rainwater, D.L., Cole, S.A., Dyer, T.D., Blangero, J., Maccluer, J.W., Mahaney, M.C., 2006, Multiple QTLs influence variation in paraoxonase 1 activity in Mexican. Americans. Hum. Biol., 78(3):341–352CrossRefGoogle Scholar
  38. Winnier, D.A., Rainwater, D.L., Cole, S.A., Williams, J.T., Dyer, T.D., Blangero, J., MacCluer, J.W., Mahaney, M.C., 2007, Sex-specific QTL effects on variation in paraoxonase 1 (PON1) activity in Mexican Americans. Genet. Epidemiol, 31(1):66–74CrossRefPubMedGoogle Scholar
  39. Yuknavage, K.L., Fenske, R.A., Kalman, D.A., Keifer, M.C., Furlong, C.E., 1997, Simulated dermal contamination with capillary samples and field cholinesterase biomonitoring. J. Toxicol. and Env. Health 51:35–55Google Scholar
  40. Zech, R., Zurcher, K., 1974, Organophosphate splitting serum enzymes in different mammals. Comp. Biochem. Physiol. 48B:427–433Google Scholar

Copyright information

© Springer 2008

Authors and Affiliations

  • C.E. Furlong
    • 1
    • 2
  • R.J Richter
    • 1
    • 2
  • W.-F. Li
    • 6
  • V.H. Brophy
    • 1
    • 2
    • 7
  • C. Carlson
    • 2
    • 8
  • M. Rieder
    • 2
  • D. Nickerson
    • 2
  • L.G. Costa
    • 3
    • 4
  • J. Ranchalis
    • 1
    • 2
  • A.J. Lusis
    • 5
  • D.M. Shih
    • 5
  • A. Tward
    • 5
    • 9
  • G.P. Jarvik
    • 1
    • 2
  1. 1.Departments of Medicine (Div. Medical Genetics)USA
  2. 2.Genome SciencesUSA
  3. 3.Environmental and Occupational Health SciencesUniversity of WashingtonSeattleUSA
  4. 4.Dept. of Human Anatomy Pharmacology and Forensic MedicineUniversity of ParmaItaly
  5. 5.Department of MedicineUCLALos AngelesUSA
  6. 6.Division of Environmental Health and Occupational MedicineNational Health Research InstitutesZhunanTaiwan
  7. 7.Roche DiagnosticsAlameda
  8. 8.The Fred Hutchinson Cancer Research CenterSeattle
  9. 9.University of CaliforniaSan Francisco

Personalised recommendations