Mammalian Genome

, Volume 6, Issue 4, pp 224–230 | Cite as

Cloning and characterization of a second human NRAMP gene on chromosome 12q13

  • S. Vidal
  • A. -M. Belouchi
  • M. Cellier
  • B. Beatty
  • P. Gros
Original Contributions


The mouse Nramp1 gene was initially identified by positional cloning as a candidate for the host resistance locus Bcg on mouse Chromosome (Chr) 1. Subsequent studies have shown that Nramp is a small gene family of at least three members in the mouse genome. We report the isolation and characterization of a cDNA clone corresponding to the second member of the human NRAMP family, NRAMP2. Predicted amino acid sequence analysis of the NRAMP2 polypeptide identifies a polytopic membrane protein highly homologous to NRAMP1, with 66% identical residues (80% overall homology), resulting in identical predicted secondary structures for the two proteins. Sequence conservation is particularly high in the predicted transmembrane domains (90%), suggesting that these regions play a key role in the structural and functional aspects common to both proteins. As opposed to its NRAMP1 counterpart, whose expression is restricted to phagocytic cells, Northern blotting analyses indicate that NRAMP2 mRNA transcripts are expressed at low levels in all tissues analyzed. Fluorescence in situ hybridization has identified 12q13 as the chromosomal location for human NRAMP2.


Northern Blotting Positional Cloning Northern Blotting Analysis Mouse Genome Resistance Locus 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Abel, L., Demenais, F. (1988). Detection of major genes for susceptibility to leprosy and its subtypes in a Caribbean island: Desirade island. Am. J. Hum. Genet. 42, 256–266.Google Scholar
  2. Bairoch, A. (1991). PROSITE: a dictionary of sites and patterns in proteins. Nucleic Acids Res. 19, 2241–2245.Google Scholar
  3. Bairoch, A. (1993). The PROSITE dictionary of sites and patterns in proteins: its current status. Nucleic Acids Res. 21, 3097–4103.Google Scholar
  4. Boyle, A.L., Feltquite, D.M., Dracopoli, N.C., Housman, D.E., Ward, D.C. (1992). Rapid physical mapping of cloned DNA on banded mouse chromosomes by fluorescence in situ hybridization. Genomics 12, 106–115.Google Scholar
  5. Bradley, D.J. (1977). Genetic control of Leishmania populations within the host. II. Genetic control of acute susceptibility of mice to L. donovani infection. Clin. Exp. Immunol. 30, 130–140.Google Scholar
  6. Bumstead, N., Barrow, P.A. (1987). Genetics of resistance to Salmonella typhimurium in newly hatched chicks. Br. Poult. Sci. 29, 521–530.Google Scholar
  7. Butler, A., Wey, A., Baker, K., and Salkoff, L. (1989). A family of putative potassium channel genes in drosophila. Science 243, 943–947.Google Scholar
  8. Cellier, M., Govoni, G., Vidal, S., Groulx, N., Liu, J., Sanchez, F., Skamene, E., Schurr, E., Gros, P. (1994). The human NRAMP gene: cDNA cloning, chromosomal mapping, genomic organization and tissue specific expression. J. Exp. Med. 180, 1741–1752.Google Scholar
  9. Dassa, E., Hofnung, M. (1985). Sequence of gene malG in E. coli K12: homologies between integral membrane components from binding protein-dependent transport systems. EMBO J. 4, 2287–2293.Google Scholar
  10. Devereux, J. (1991). The GCG sequence analysis softward package, version 7.0. Genetics Computer Group, Inc., University Research Park, Madison, Wisconsin, USA.Google Scholar
  11. Dosik, J.K., Barton, C.H., Holiday, D.L., Krall, M.M., Blackwell, J.M., Mock, B.A. (1994). An Nramp-related sequence maps to mouse Chromosome 17. Mamm. Genome 5, 458–460.Google Scholar
  12. Feinberg, A.P., Vogelstein, B. (1984). A technique for radiolabelling DNA restriction endonucleases fragments to high specific activity. Anal. Biochem. 132, 6–13.Google Scholar
  13. Govoni, G., Vidal, S., Cellier, M., Lepage, P., Malo, D., Gros, P. (1994). Structural organization and cell specific expression of the mouse Nramp1 gene. Genomics (in press).Google Scholar
  14. Gros, P., Skamene, E., Forget, A. (1981). Genetic control of natural resistance to Mycobacterium bovis BCG. J Immunol. 127, 2417–2421.Google Scholar
  15. Gros, P., Skamene, E., Forget, A. (1983). Cellular mechanisms of genetically-controlled host resistance to Mycobacterium bovis (BCG). J. Immunol. 131, 1966–1973.Google Scholar
  16. Gruenheid, S., Cellier, M., Vidal, S., Gros, P. (1994). Identification and characterization of a second mouse Nramp gene. Genomics, in press.Google Scholar
  17. Hart, C.E., Ou, C.H., Galpin, J.C., Moore, J., Bacheler, L.T., Wasmuth, J.J., Petteway, S.R. Jr., Schochetman, G. (1990). Human chromosome 12 is required for elevated HIV-1 expression in human-hamster hybrid cells. Science 246, 488–491.Google Scholar
  18. Heng, H., Tsui, L-C. (1993). Models of DAPI banding and simultaneous in situ hybridization. Chromosoma 102, 325–332.Google Scholar
  19. Kerppola, R.E., Ames, G.F.-L. (1992). Topology of the hydrophobic membrane-bound components of the histidine periplasmic permease. Comparison with other members of the family. J. Biol. Chem. 267, 2329.Google Scholar
  20. Kishimoto, A., Nishiyama, K., Nalanishi, H., Uratsuji, Y., Nomura, H., Takeyama, Y., Nishizuka, Y. (1985). Studies on the phosphorylation of myelin basic proteins by protein kinase C and adenosine 3′:5′-monophosphate-dependent protein kinase. J. Biol. Chem. 260, 12492–12499.Google Scholar
  21. Kyte, J., Doolittle, R.F. (1982). A simple method for displaying the hydropathy of a protein. J. Mol. Biol. 157, 105.Google Scholar
  22. Lichter, P., Tang, C.J.C., Call, K., Hermanson, G., Evans, G.A., Housman, D., Ward, D. (1990). High-resolution mapping of chromosome 11 by in situ hybridization with cosmid clones. Science 247, 64–69.Google Scholar
  23. Lurie, M.B. (1964). Resistance to Tuberculosis: Experimental Studies in Native and Acquired Defensive Mechanisms. (Cambridge, Mass.: Harvard University Press).Google Scholar
  24. Lurie, M.B., Dannenberg, A.M. (1965). Macrophage function in infectious disease with inbred rabbits. Bacteriol. Rev. 29 466.Google Scholar
  25. Malo, D., Vidal, S., Skamene, E., Gros, P. (1993a). High resolution linkage map in the vicinity of the host resistance locus Beg on mouse chromosome 1. Genomics, 16, 655–663.Google Scholar
  26. Malo, D., Vidal, S.M., Leiman, J., Ward, D., Gros, P. (1993b). Physical delineation of the minimal chromosomal segment encompassing the host resistance locus Bcg. Genomics 17, 667–675.Google Scholar
  27. Malo, D., Vogan, K., Vidal, S., Hu, J., Cellier, M., Schurr, E., Fuks, A., Morgan, K., Gros, P. (1994). Haplotype mapping and sequence analysis of the mouse Nramp gene predicts susceptibility to infection with intracellular parasites. Genomics 23, 51–61.Google Scholar
  28. Nathan, C., Xie, Q.-W. (1994). Nitric oxide synthase: roles, tolls, and controls. Cell, 78, 915–918.Google Scholar
  29. Newstein, M., Stanbridge, E.J., Casey, G., Shank, P.R. (1990). Human chromosome 12 encodes a species-specific factor which increases human immunodeficiency virus type 1 tat-mediated trans activation in rodent cells. J. Virol. 64, 4565–4567.Google Scholar
  30. Ottenhoff, T.H., de Vries, R.R. (1987). HLA class II immune response and suppression genes in leprosy. Int. J. Lepr. 55, 521–534.Google Scholar
  31. Pinna, L.A. (1990). Casein kinase 2: an “eminence grise” in cellular regulation? Biochim. Biophys. Acta 1054, 267–284.Google Scholar
  32. Plant, J.E., Glynn, A. (1976). Genetics of resistance to infection with Salmonella typhimurium in mice. J. Infect. Dis. 133, 72–78.Google Scholar
  33. Sambrook, J., Fritsch, E.F., Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual, 2nd ed. (Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory Press).Google Scholar
  34. Sanger, F., Nicklen, S., Coulson, A.R. (1977). DNA sequencing with chain termination inhibitors. Proc. Natl. Acad. Sci. USA 74: 5463–5467.Google Scholar
  35. Saurin, W., Koster, W., Dassa, E. (1994). Bacterial binding protein-dependent permeases: characterization of distinctive signatures for functionally related integral cytoplasmic membrane proteins. Mol. Microbiol. 12, 993–1004.Google Scholar
  36. Schurr, E., Skamene, E., Morgan, K., Chu, M.-L., Gros, P. (1990). Mapping of Col3a1 and Col6a3 to proximal murine chromosome 1 identifies conserved linkage of structural protein genes between murine chromosome 1 and human chromosome 2q. Genomics 8, 477–486.Google Scholar
  37. Schurr, E., Moorage, K., Gros, P., Skamene, E. (1991). Genetics of leprosy. Am. J. Trop. Med. Hyg. 44, 4–11.Google Scholar
  38. Shields, E.D., Russell, D.A., Pericak-Vance, M.A. (1987). Genetic epidemiology of the susceptibility to leprosy. J. Clin. Invest. 79, 1139–1143.Google Scholar
  39. Skamene, E., Gros, P., Forget, A., Kongshavn, P.A.L., St.-Charles, C., Taylor, B.A. (1982). Genetic regulation of resistance to intracellular pathogens. Nature 297, 506–509.Google Scholar
  40. Stead, W.W. (1992). Genetics and resistance to tuberculosis. Could resistance be enhanced by genetic engineering? Ann. Intern. Med. 116, 937–941.Google Scholar
  41. Unkles, S.E., Hawker, K.L., Grieve, C., Campbell, E.I., Montague, P., Kinghorn, J.R. (1991). crnA encodes a nitrate transporter in Aspergillus nidulans. Proc. Natl. Acad. Sci. USA 88, 204–208.Google Scholar
  42. Vidal, S., Malo, D., Vogan, K., Skamene, E., Gros, P. (1993). Natural resistance to infection with intracellular parasites: isolation of a candidate for Bcg. Cell 73, 469–486.Google Scholar
  43. von Heijne, G. (1986). The distribution of positively charged residuces in bacterial inner membrane proteins correlates with the trans-membrane topology. EMBO J. 5, 3021–3027.Google Scholar
  44. Woodgett, J.R., Gould, K.L., Hunter, T. (1986). Substrate specificity of protein kinase C. Use of synthetic peptides corresponding to physiological sites as probes for substrate recognition requirements. Eur. J. Biochem. 161, 177–184.Google Scholar

Copyright information

© Springer-Verlag New York Inc 1995

Authors and Affiliations

  • S. Vidal
    • 1
    • 2
  • A. -M. Belouchi
    • 1
  • M. Cellier
    • 1
  • B. Beatty
    • 3
  • P. Gros
    • 1
    • 2
  1. 1.Department of BiochemistryMcGill UniversityMontréalCanada
  2. for the Study of Host ResistanceMcGill UniversityMontréalCanada
  3. 3.CGAT Fish Mapping Resource Center, Department of PathologyThe Hospital for Sick ChildrenTorontoCanada

Personalised recommendations