Isolation and characterization of BrMDR1 a novel MDR-type ATP-binding cassette (ABC) transporter in Brassica rapa L.

  • Lee, Sun-Yong (Department of Horticulture, Hankyong National University) ;
  • Jung, Yu-Jin (Department of Horticulture, Hankyong National University) ;
  • Kang, Kwon-Kyoo (Department of Horticulture, Hankyong National University)
  • Published : 2009.06.30

Abstract

A cDNA clone encoding a MDR-like ABC transporter protein was isolated from Brassica rapa seedlings, through rapid amplification of cDNA ends (RACE). This gene (named as Brmdr 1; GenBank accession no.: DQ296184 ) had a total length of 4222 bp with an open reading frame of 3900 bp, and encoded a predicted polypeptide of 1300 amino acids with a molecular weight of 143.1 kDa. The BrMDR1 protein shared 71.0, 62.5, 60.0 and 58.2% identity with other MDR proteins isolated from Arabidopsis thaliana (AAN28720), Coptis japonica (CjMDR), Gossypium hirsutum (GhMDR) and Triticum aestivum (TaMDR) at amino acid level, respectively. Southern blot analysis showed that Brmdr1 was a low-copy gene. Expression pattern analysis revealed that Brmdr1 constitutively expressed in the root, stem petals and stamens, but with lower expression in leaves and open flowers. The domains analysis showed that BrMDR1 protein possessed two transmembrane domains (TMDs) and two nucleotide binding domains (NBDs) arranging in "TMD1-NBD1-TMD2-NBD2" direction, which is consistent with other MDR transporters. Within NBDs three characteristic motifs common to all ABC transporters, "Walker A", "Walker B" and C motif, were found. These results indicate that BrMDR1 is a MDR-like ABC transporter protein that may be involved in the transport and accumulation of secondary metabolites.

Keywords

References

  1. Bouwmeester, H.J., R. Matusova, S. Zhongkui and M.H. Beale. 2003. Secondary metabolite signalling in host-parasitic plant interactions. Curr Opin Plant Biol, 6: 358-364 https://doi.org/10.1016/S1369-5266(03)00065-7
  2. Geisler, M., J.J. Blakeslee, R. Bouchard, O.R. Lee, V. Vincenzetti, A. Bandyopadhyay, B. Titapiwatanakun, W.A. Peer, A. Bailly, E.L. Richards, K.F. Ejendal, A.P. Smith, C. Baroux, U. Grossniklaus, A. Muller, C.A. Hrycyna, R. Dudler, A.S. Murphy and E. Martinoia, 2005. Cellular efflux of auxin catalyzed by the Arabidopsis MDR/PGP transporter AtPGP1. Plant J. 44: 179-194 https://doi.org/10.1111/j.1365-313X.2005.02519.x
  3. Geisler, M., H.U. Kolukisaoglu, R. Bouchard, K. Billion, J. Berger, B. Saal, N. Frangne, Z. Koncz-Kalman, C. Koncz, R. Dudler, J.J. Blakeslee, A.S. Murphy, E. Martinoia and B. Schulz. 2003. TWISTED DWARF1, a unique plasma membrane-anchored immunophilin-like protein, interacts with Arabidopsis multidrug resistance-like transporters AtPGP1 and AtPGP19. Mol Biol Cell 14 : 4238-4249 https://doi.org/10.1091/mbc.E02-10-0698
  4. Geourjon, C. and G. Deleage. 1995. SOPMA: Significant improvements in protein secondary structure prediction by consensus prediction from multiple alignments. Comput Appl Biosci 11: 681-684
  5. Harborne, J.B. 1990. Role of secondary metabolites in chemical defence mechanisms in plants. Ciba Found Symp 154: 126-134
  6. Hughes, E.H. and J.V. Shanks. 2002. Metabolic engineering of plants for alkaloid production. Metab Eng 4 : 41-48 https://doi.org/10.1006/mben.2001.0205
  7. Jacobs, D.I., M. Gaspari, J. van der Greef, R. van der Heijden and R. Verpoorte. 2005. Proteome analysis of the medicinal plant Catharanthus roseus. Planta 221: 690-704 https://doi.org/10.1007/s00425-004-1474-4
  8. Jasinski, M., E. Ducos, E. Martinoia and M.Boutry. 2003. The ATP-binding cassette transporters: Structure, function, and gene family comparison between rice and Arabidopsis. Plant Physiol 131:1169-1177 https://doi.org/10.1104/pp.102.014720
  9. Jordan, MA, D. Thrower and L. Wilsen. 1991. Mechanism of inhibition of cell proliferation by Vinca alkaloids. Cancer Res 51: 2212-2222
  10. Kumar, S.K., K. Tamura, I.B. Jakobsen and M. Nei. 2001. MEGA2: Molecular evolutionary genetics analysis software. Bioinformatics 17: 1244-1245 https://doi.org/10.1093/bioinformatics/17.12.1244
  11. Loo, T.W. and D.M. Clarke. 2005. Recent progress in understanding the mechanism of P-glycoprotein-mediated drug efflux. J Membr Biol 206: 173-185 https://doi.org/10.1007/s00232-005-0792-1
  12. Majse N., E.S. Ida, G.H. Bjarne, G.F. Fernando, H.N.E. Hussam, H.H.N. Morten, B.J. Niels, L. Jing and A.H. Barbara. 2006. Cytochromes P450 in the biosynthesis of glucosinolates and indole alkaloids. Phytochemistry 5: 331-346 https://doi.org/10.1007/s11101-006-9004-6
  13. Morant, M., S. Bak, B.L. Moller. And D. Werck-Reichhart. 2003. Plant cytochrommes 450: Tools for pharmacology, plant protection and phytoremediation. Curr Opin Biotechnol 14: 151-162 https://doi.org/10.1016/S0958-1669(03)00024-7
  14. Oksman-Caldentey, K. M. and D. Inze. 2004. Plant cell factories in the post-genomic era: New ways to produce designer secondary metabolites. Trends Plant Sci 9: 433-440 https://doi.org/10.1016/j.tplants.2004.07.006
  15. Saitou, N. and M. Nei. 1987. The neighbor-joining method: A new method for reconstructing phylogenetic trees. Mol Biol Evol 4: 406-425
  16. Sambrook, J., E.F. Fritsch.and T. Maniatis. 1989. Molecular Cloning: A Laboratory Manua Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
  17. Santelia, D., V. Vincenzetti, E. Azzarello, L. Bovet, Y. Fukao, P. Duchtig, S. Mancuso, E. Martinoia and M. Geisler. 2005. MDR-like ABC transporter AtPGP4 is involved in auxinmediated lateral root and root hair development. FEBS Lett 579:5399-5406 https://doi.org/10.1016/j.febslet.2005.08.061
  18. Sato, H., Y. Kobayashi, H. Fukui and M. Tabata. 1990. Specific differences in tolerance to exogenous berberine among plant cell cultures. Plant Cell Rep 9:133-136 https://doi.org/10.1007/BF00232088
  19. Schulz, B. and H.U. Kolukisaoglu. 2006. Genomics of plant ABC transporters: The alphabet of photosynthetic life forms or just holes in membranes? FEBS Lett 580:1010-1016 https://doi.org/10.1016/j.febslet.2006.01.002
  20. Shitan, N., I. Bazin, K. Dan, K. Obata, K. Kigawa, K. Ueda, F. Sato, C. Forestier and K. Yazaki. 2003. Involvement of CjMDR1, a plant MDR-type ABC protein, in alkaloid transport in Coptis japonica. Proc Natl Acad Sci USA 100: 751-756 https://doi.org/10.1073/pnas.0134257100
  21. Terasaka, K., J.J. Blakeslee, B. Titapiwatanakun, W.A. Peer, A. Bandyopadhyay, S.N. Makam, O.R. Lee, E.L. Richards, A.S. Murphy, F. Sato and K. Yazaki. 2005. PGP4, an ATP binding cassette P-glycoprotein, catalyzes auxin transport in Arabidopsis thaliana roots. Plant Cell 17:2922-2939 https://doi.org/10.1105/tpc.105.035816
  22. Theodoulou, F. L. 2000. Plant ABC transporters. Biochim Biophys Acta 1465: 79-103 https://doi.org/10.1016/S0005-2736(00)00132-2
  23. Windsor, B., S.J. Roux and A. Lloyd. 2003. Multiherbicide tolerance conferred by AtPgp1 and apyrase overexpression in Arabidopsis thaliana. Nat Biotechnol 21: 428-433 https://doi.org/10.1038/nbt809
  24. Yazaki, K. 2005 Transporters of secondary metabolites. Curr Opin Plant Biol 8: 301-307 https://doi.org/10.1016/j.pbi.2005.03.011
  25. Yazaki, K. 2006. ABC transporters involved in the transport of plant secondary metabolites. FEBS Lett 580: 1183-1191 https://doi.org/10.1016/j.febslet.2005.12.009
  26. Zhao, J., L.C. Davis and R. Verpoorte. 2005. Elicitor signal transduction leading to production of plant secondary metabolites. Biotechnol Adv 23: 283-333 https://doi.org/10.1016/j.biotechadv.2005.01.003