The macrophage-specific membrane protein Nramp controlling natural resistance to infections in mice has homologues expressed in the root system of plants Research article Received: 20 June 1995 Accepted: 13 September 1995 DOI:
Cite this article as: Belouchi, A., Cellier, M., Kwan, T. et al. Plant Mol Biol (1995) 29: 1181. doi:10.1007/BF00020461 Abstract
In mice, natural resistance or susceptibility to infection with
Mycobacteria, Salmonella, and Leishmania is controlled by a gene named Bcg. Bcg regulates the capacity of macrophages to limit intracellular replication of the ingested parasites, and is believed to regulate a key bactericidal mechanism of this cell. Recently, we have cloned the Bcg gene and shown that it encodes a novel macrophage-specific membrane protein designated Nramp. A routine search of the public databases for sequences homologous to Nramp identified 3 expressed sequence tags (EST) that show strong similarities to the mammalian protein.
We report the identification and cloning of a full-length cDNA clone corresponding to a plant homologue (
OsNramp1) of mammalian Nramp. Predicted amino acid sequence analysis of the plant protein indicates a remarkable degree of similarity (60% homology) with its mammalian counterpart, including identical number, position, and composition of transmembrane domains, glycosylation signals, and consensus transport motif, suggesting an identical overall secondary structure and membrane organization for the two proteins. This high degree of structural similarity indicates that the two proteins may be functionally related, possibly through a common mechanism of transport. RNA hybridization studies and RT-PCR analyses indicate that OsNramp1 mRNA is expressed primarily in roots and only at very low levels in leaves/stem. DNA hybridization studies indicate that OsNramp1 is not a single gene, but rather forms part of a novel gene family which has several members in all plants tested including cereals such as rice, wheat, and corn, and also in common weed species. The striking degree of conservation between the macrophage-specific mammalian Nramp and its OsNramp1 plant homologue is discussed with respect to possible implications in the metabolism of nitrate in both organisms. Key words gene family rice root-specific gene expression membrane protein Crna nitrate transport References
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ: Basic local alignment search tool. J Mol Biol 215: 403–410 (1990).
Anderson MP, Gregory RJ, Thompson S, Souza DW, Paul S, Mulligan RC, Smith AE, Welch MJ: Demonstration that CFTR is a Cl
channel by alteration of its anion selectivity. Science 253: 202–205 (1991).
Aslam M, Travis RL, Huffaker RC: Comparative kinetics and reciprocal inhibition of nitrate and nitrite uptake in roots of uninduced and induced barley seedlings. Plant Physiol 99: 1124–1133 (1992).
Bairoch A: The PROSITE dictionary of sites and patterns in proteins: its current status. Nucl Acids Res 21: 3097–4103 (1993).
Barton CH, White JK, Roach TIA, Blackwell JM: NH
-terminal sequence of a macrophage-expressed natural resistance-associated macrophage protein (
) encodes a proline/serine-rich putative
homology 3-binding domain. J Exp Med 179: 1683–1687 (1994).
Cellier M, Govoni G, Vidal SM, Kwan T, Groulx N, Liu J, Sanchez F, Skamene E, Schurr E, Gros P: Human natural resistance-associated macrophage protein: cDNA cloning, chromosomal mapping, genomic organization, and tissue-specific expression. J Exp Med 180: 1741–1752 (1994).
Chirgwin JM, Prybyla AA, MacDonald RJ, Rutter WJ: Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18: 5294–5299 (1979).
Crawford NM, Arst Jr HN: The molecular genetics of nitrate assimilation in fungi and plants. Annu Rev Genet 27: 115–146 (1993).
Dassa E, Hofnung M: Sequence of gene MalG in E. Coli K12: Homologies between integral membrane components from binding protein-dependent transport systems. EMBO J 4: 2287–2293 (1985).
Devereux J: The GCG sequence analysis software package, version 7.0. Genetics Computer Group, University Reseaarch Park, Madison, WI (1991).
Doddema H, Telkamp GP: Uptake of nitrate by mutants of
, disturbed in uptake or reduction of nitrate II kinetics. Physiol Plant 45: 332–338 (1979).
Eisenberg D, Schwarz E, Komaromy M, Wall R: Analysis of membrane and surface protein sequences with the hydrophobic moment plot. J Mol Biol 179: 125–142 (1984).
Epstein DJ, Vekemans M, Gros P:
, a mutation affecting development of the mouse neural tube, shows a deletion within the paired homeodomain of Pax-3. Cell 67: 767–774 (1991).
Feinberg AP, Vogelstein BA: technique for radiolabelling DNA restriction endonucleases fragments to high specific activity. Anal Biochem 132: 6–13 (1984).
Glass AD, Shaff JE, Kochian LV: Studies of the uptake of nitrate in barley. IV. Electrophysiology. Plant Physiol 99: 456–463 (1992).
Goto Y, Buschman E, Skamene E: Regulation of host resistance to
Mycobacterium intracellulare in vivo
gene. Immunogenetics 30: 218–221 (1989).
Green S, Nacy CA, Meltzer M: Cytokine-induced synthesis of nitrogen oxides in macrophages: a protective host response to
and other intracellular pathogens. J Leuk Biol 50: 93–103 (1991).
Gros P, Skamene E, Forget A: Genetic control of natural resistance to
BCG. J Immunol 127: 2417–2421 (1981).
Gros P, Skamene E, Forget A: Cellular mechanisms of genetically-controlled host resistance to
(BCG). J Immunol 131: 1966–1973 (1983).
Gruenheid S, Cellier M, Vidal SM, Gros P: Identification and characterization of a second mouse
gene. Genomics 25: 514–525 (1995).
Hole D, Emran AM, Fares Y, Drew MC: Induction of nitrate transport in maize roots, and kinetics of influx, measured with nitrogen-13. Plant Physiol 93: 642–647 (1990).
Jacobsen KS, Breivold E, Hornes E: Purification of mRNA directly from crude plant tissues in 15 minutes using magnetic oligo dT microspheres. Nucl Acids Res 18: 3669 (1990).
Kerppola RE, Ames GF-L: Topology of the hydrophobic membrane-bound components of the histidine periplasmic permease. Comparison with other members of the family. J Biol Chem 267: 2329–2336 (1992).
Kieferle S, Fong P, Bens M, Vandewalle A, Jentsch TJ: Two highly homologous members of the CIC chloride channel family in both rat and human kidney. Proc Natl Acad Sci USA 91: 6943–6947 (1994).
Kyte J, Doolittle RF: A simple method for displaying the hydropathy of a protein. J Mol Biol 157: 105–132 (1982).
Lissner CR, Swanson RN, O'Brien AD: Genetic control of the innate resistance of mice to
: expression of the
gene in peritoneal and splenic macrophages isolated
. J Immunol 131: 3006–3013 (1983).
Malo D, Schurr E, Epstein DJ, Vekemans M, Skamene E, Gros P: The host resistance locus
is tightly linked to a group of cytoskeleton-associated protein genes that include villin and desmin. Genomics 10: 356–364 (1991).
Malo D, Vidal SM, Hu J, Skamene E, Gros P: High resolution linkage map in the vicinity of the host resistance locus
. Genomics 16: 655–663 (1993).
Malo D, Vidal SM, Lieman JH, Ward DC, Gros P: Physical delineation of the minimal chromosomal segment encompassing the murine host resistance locus
. Genomics 17: 667–675 (1993).
Malo D, Vogan K, Vidal SM, Hu J, Cellier M, Schurr E, Fuks A, Morgan K, Gros P: Haplotype mapping and sequence analysis of the mouse
gene predict susceptibility to infection with intracellular parasites. Genomics 23: 51–61 (1994).
Mangalan H: Striding the turf of the gang of four. Trends Genet 18: 187–188 (1993).
Nathan C: Nitric oxide as a secretory product of mammalian cells. FASEB J 6: 3051–3061 (1992).
Nathan CF, Hibbs JB: Role of nitric oxyde synthesis in macrophage antimicrobial activity. Curr Opin Immunol 3: 65–70 (1991).
Noonan KE, Rominson IB: mRNA phenotyping by enzymatic amplification of randomly primed cDNA. Nucl Acids Res 16: 10366 (1988).
Ruiz-Cristin J, Briskin DP: Characterization of a H
symport associated with plasma membrane vesicles of maize roots using
as a radiotracer analog. Arch Biochem Biophys 285: 74–82 (1991).
Sahin-Toth M, Dunten RL, Gonzales A, Kaback HR: Functional interactions between putative intramembrane charged residues in the lactose permease of
. Proc Natl Acad Sci USA 89: 10547–10551 (1992).
Sambrook J, Fritsch EF, Maniatis T: ‘Molecular Cloning: A Laboratory Manual’, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).
Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain termination inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).
Saurin W, Koster W, Dassa E: Bacterial binding protein-dependent permeases: characterization of distinctive signatures for functionally related integral cytoplasmic membrane proteins. Mol Microbiol 12: 993–1004 (1994).
Siddiqi MY, Glass ADM, Ruth TJ, Fernando M: Studies on the regulation of nitrate influx by barley using
. Plant Physiol 90: 806–813 (1989).
Siddiqi MY, Glass ADM, Ruth TJ, Rufty TW: Studies of the uptake of nitrate in barley. Plant Physiol 93: 1426–1432 (1990).
Skamene E, Pietrangeli CE: Genetics of the immune response to infectious pathogens. Curr Onin Immunol 3: 511–517 (1991).
Stach JL, Gros P, Forget A, Skamene E: Phenotypic expression of genetically controlled natural resistance to
(BCG) J Immunol 132: 888–892 (1984).
Stuehr DJ, Nathan CF: Nitric oxide. A macrophage product responsible for cytostasis and respiratory inhibition in tumor target cells. J Exp Med 169: 1543–1555 (1989).
Tsay Y-F, Schroeder JI, Feldmann KA, Crawford NM: The herbicide sensitivity gene
encodes a nitrate-inducible nitrate transporter. Cell 72: 705–713 (1993).
Unkles SE, Hawker KL, Grieve C, Campbell EI, Van Montague P, Kinghorn JR:
encodes a nitrate transporter in
. Proc Natl Acad Sci USA 88: 204–208 (1991).
Vidal SM, Malo D, Vogan K, Skamene E, Gros P: Natural resistance to infection with intracellular parasites: Isolation of a candidate for
. Cell 73: 469–485 (1993).
Vidal SM, Tremblay M, Govoni G, Gauthier S, Sebastiani G, Malo D, Olivier M, Skamene E, Jothy S, Gros P: The
Ity/Lsh/Bcg locus: natural resistance to infection with intracellular parasites is abrogated by disruption of the Nramp1 gene. J Exp Med, in press (1995). Copyright information
© Kluwer Academic Publishers 1995