Advertisement

Molecular Biology Reports

, Volume 38, Issue 4, pp 2679–2684 | Cite as

Genotype and allele frequencies of divalent metal transporter 1 polymorphism in Turkish population

  • Zeliha KayaaltiEmail author
  • Miyase Odabaşi
  • Tülin Söylemezoğlu
Article

Abstract

Divalent metal transporter 1 (DMT1 also known as DCT1, NRAMP2 or SCL11A2) is a membrane-bound divalent metal transporter which is conserved from prokaryotes to higher eukaryotes. It has been postulated to play important roles in intestinal iron absorption at the brush border of duodenal enterocytes, erythroid iron utilization, hepatic iron accumulation, placental iron transfer, and other processes. DMT1 gene which contains at least four isoforms (1A/+IRE, 1A/−IRE, 2/+IRE and 2/−IRE) is located on chromosome 12q13 in human. DMT1 mediates the transport of a wide range of metals, including the essential metals Fe2+, Zn2+, Mn2+, Cu2+, Co2+, Ni2+ and toxic metals such as Cd2+ and Pb2+. The intention of this study is to determine that IVS4+44C/A single nucleotide polymorphism in DMT1 gene of Turkish population. For this purpose blood samples from 192 female and 192 male volunteers were analyzed. DMT1 gene was amplified with the polymerase chain reaction-restriction fragment length polymorphism technique and 351 bp oligonucleotide was produced. The amplified oligonucleotides were cut with MnlI restriction enzyme according to their polymorphic characteristics. Digested and undigested products were separated on a 2% agarose gel electrophoresis, visualized by ethidium bromide staining under an ultraviolet illuminator. The genotype frequencies of DMT1 IVS4+44C/A polymorphism were determined as 47.9% for CC, 40.1% for AC and 12.0% for AA genotypes. The frequency of the C allele was found to as 68.0% and of the A allele as 32.0%. The genotype frequencies were consistent with Hardy–Weinberg equilibrium (χ2 = 2.394; Exact P = 0.128).

Keywords

Single nucleotide polymorphism Divalent metal transporter 1 Turkish population Restriction enzyme 

Abbreviations

AREs

Antioxidant responsive elements

Dntp

Dinucleotide triphosphate

PCR

Polymerase chain reaction

RFLP

Restriction fragment length polymorphism

SNP

Single nucleotide polymorphism

DMT1

Divalent metal transporter 1

DCT1

Divalent cation transporter 1

NRAMP2

Natural resistance-associated macrophage protein 2

SCL11A2

Solute carrier family 11

+IRE

With iron responsive element

−IRE

Without iron responsive element

IVS4+44C/A

Intervening sequence 4+44

References

  1. 1.
    Andrews NC, Levy JE (1998) Iron is hot: an update on the pathophysiology of hemochromatosis. Blood 92:1845–1851PubMedGoogle Scholar
  2. 2.
    Li W, Hellsten A, Nyhalah JD, Yuan XM (2004) Enhanced expression of natural resistance-associated macrophage protein 1 in atherosclerotic lesions may be associated with oxidized lipid-induced apoptosis. Ann N Y Acad Sci 1030:202–207PubMedCrossRefGoogle Scholar
  3. 3.
    Bartzokis G, Sultzer D, Mintz J, Holt LE, Marx P, Phelan CK, Marder SR (1994) In vivo evaluation of brain iron in Alzheimer’s disease and normal subjects using MRI. Biol Psychiatry 35:480–487PubMedCrossRefGoogle Scholar
  4. 4.
    Bartzokis G, Cummings J, Perlman S, Hance DB, Mintz J (1999) Increased basal ganglia iron levels in Huntington disease. Arch Neurol 56:569–574PubMedCrossRefGoogle Scholar
  5. 5.
    Dexter DT, Wells FR, Lees AJ, Agid F, Agid Y, Jenner P, Marsden CD (1989) Increased nigral iron content and alterations in other metal ions occurring in brain in Parkinson’s disease. J Neurochem 52:1830–1836PubMedCrossRefGoogle Scholar
  6. 6.
    Nelson N (1999) Metal ion transporters and homeostasis. EMBO J 8:4361–4371CrossRefGoogle Scholar
  7. 7.
    Fleming MD, Romano MA, Su MA, Garrick LM, Garrick MD, Andrews NC (1998) Nramp2 is mutated in the anemic Belgrade (b) rat: evidence of a role for Nramp2 in endosomal iron transport. Proc Natl Acad Sci USA 95:1148–1153PubMedCrossRefGoogle Scholar
  8. 8.
    Andrews NC (1999) The iron transporter DMT1. Int J Biochem Cell Biol 31:991–994PubMedCrossRefGoogle Scholar
  9. 9.
    Wessling-Resnick M (2000) Iron transport. Annu Rev Nutr 20:129–151PubMedCrossRefGoogle Scholar
  10. 10.
    Garrick MD, Singleton ST, Vargas F, Kuo HC, Zhao L, Knöpfel M, Davidson T, Costa M, Paradkar P, Roth JA, Garrick LM (2006) DMT1: which metals does it transport? Biol Res 39:79–85PubMedCrossRefGoogle Scholar
  11. 11.
    Cellier M, Prive G, Belouchi A, Kwan T, Rodrigues V, Chia W, Gros P (1995) Nramp defines a family of membrane proteins. Proc Natl Acad Sci USA 92:10089–10093PubMedCrossRefGoogle Scholar
  12. 12.
    Hubert N, Hentze MW (2002) Previously uncharacterized isoforms of divalent metal transporter (DMT)-1: implications for regulation and cellular function. Proc Natl Acad Sci USA 99:12345–12350PubMedCrossRefGoogle Scholar
  13. 13.
    Gunshin H, Mackenzie B, Berger UV, Gunshin Y, Romero MF, Boron WF, Nussberger S, Gollan JL, Hediger MA (1997) Cloning and characterization of a mammalian proton-coupled metal-ion transporter. Nature 388:482–488PubMedCrossRefGoogle Scholar
  14. 14.
    Lee PL, Gelbart T, West C, Halloran C, Beutler E (1998) The human Nramp2 gene: characterization of the gene structure, alternative splicing, promoter region and polymorphisms. Blood Cells Mol Dis 24:199–215PubMedCrossRefGoogle Scholar
  15. 15.
    Canonne-Hergaux F, Gruenheid S, Ponka P, Gros P (1999) Cellular and subcellular localization of the Nramp2 iron transporter in the intestinal brush border and regulation by dietary iron. Blood 93:4406–4417PubMedGoogle Scholar
  16. 16.
    Fairweather-Tait SJ, Harvey L, Heath ALM, Soe M (2007) Effect of SNPs on iron metabolism. Genes Nutr 2:15–19PubMedCrossRefGoogle Scholar
  17. 17.
    Mims MP, Guan Y, Pospisilova D, Priwitzerova M, Indrak K, Ponka P, Divoky V, Prchal JT (2005) Identification of a human mutation of DMT1 in a patient with microcytic anemia and iron overload. Blood 105:1337–1342PubMedCrossRefGoogle Scholar
  18. 18.
    Howitt J, Putz U, Lackovic J, Doan A, Dorstyn L, Cheng H, Yang B, Chan-Ling T, Silke J, Kumar S, Tan SS (2009) Divalent metal transporter 1 (DMT1) regulation by Ndfip1 prevents metal toxicity in human neurons. Proc Natl Acad Sci USA 106:15489–15494PubMedCrossRefGoogle Scholar
  19. 19.
    Iacoviello L, Ciccarone E, Donati MB (2002) The genetics of macrovascular disease in diabetes. Br J Diabetes Vasc Dis 2:364–368CrossRefGoogle Scholar
  20. 20.
    Kelleher T, Ryan E, Barrett S, O’Keane C, Crowe J (2004) DMT1 genetic variability is not responsible for phenotype variability in hereditary hemochromatosis. Blood Cells Mol Dis 33:35–39PubMedCrossRefGoogle Scholar
  21. 21.
    Henle ES, Linn S (1997) Formation, prevention, and repair of DNA damage by iron/hydrogen peroxide. J Biol Chem 272:19095–19098PubMedCrossRefGoogle Scholar
  22. 22.
    O’Riordan DK, Debnam ES, Sharp PA, Simpson RJ, Taylor EM, Srai SK (1997) Mechanisms involved in increased iron uptake across rat duodenal brush-border membrane during hypoxia. J Physiol 500:379–384PubMedGoogle Scholar
  23. 23.
    Gunshin H, Fujiwara Y, Custodio AO, Direnzo C, Robine S, Andrews NC (2005) Slc11a2 is required for intestinal iron absorption and erythropoiesis but dispensable in placenta and liver. J Clin Invest 115:1258–1266PubMedGoogle Scholar
  24. 24.
    Priwitzerova M, Nie G, Sheftel AD, Pospisilova D, Divoky V, Ponka P (2005) Functional consequences of the human DMT1 (SLC11A2) mutation on protein expression and iron uptake. Blood 106:3985–3987PubMedCrossRefGoogle Scholar
  25. 25.
    Iolascon A, d’Apolito M, Servedio V, Cimmino F, Piga A, Camaschella C (2006) Microcytic anemia and hepatic iron overload in a child with compound heterozygous mutations in DMT1 (SCL11A2). Blood 107:349–354PubMedCrossRefGoogle Scholar
  26. 26.
    Beaumont C, Delaunay J, Hetet G, Grandchamp B, de Montalembert M, Tchernia G (2006) Two new human DMT1 gene mutations in a patient with microcytic anemia, low ferritinemia, and liver iron overload. Blood 107:4168–4170PubMedCrossRefGoogle Scholar
  27. 27.
    Fleming MD, Trenor CC 3rd, Su MA, Foernzler D, Beier DR, Dietrich WF, Andrews NC (1997) Microcytic anaemia mice have a mutation in Nramp2, a candidate iron transporter gene. Nat Genet 16:383–386PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Zeliha Kayaalti
    • 1
    Email author
  • Miyase Odabaşi
    • 1
  • Tülin Söylemezoğlu
    • 1
  1. 1.Institute of Forensic MedicineAnkara UniversityAnkaraTurkey

Personalised recommendations