, Volume 226, Issue 4, pp 1031–1039

AtMTM1, a novel mitochondrial protein, may be involved in activation of the manganese-containing superoxide dismutase in Arabidopsis

  • Zhao Su
  • Mao-Feng Chai
  • Ping-Li Lu
  • Rui An
  • Jia Chen
  • Xue-Chen Wang
Original Paper


Mtm1p is essential for the posttranslational activation of manganese-containing superoxide dismutase (SOD2) in Saccharomyces cerevisiae; however, whether the same holds true for Arabidopsis thaliana is unknown. In this study, by using the yeast mtm1 mutant complementation method, we identified a putative MTM gene (AtMTM1, At4g27940) that is necessary for SOD2 activation. Further, analysis of SOD activity revealed that an SOD2 defect is rescued in the yeast mutant Y07288 harboring the AtMTM1 gene. Related mRNA-level analysis showed the AtMTM1 gene is induced by paraquat but not by hydrogen peroxide, which indicates that this gene is related to the superoxide scavenger SOD. In addition, an AtMTM1::GFP fusion construct was transiently expressed in the protoplasts, and it was localized to the mitochondria. Furthermore, sequence deletion analysis of AtMTM1 revealed that the code region (amino acid (aa) 60–198) of Mtm1p plays an important role in localization of the protein to the mitochondria. Regulation of AtMTM1 gene expression was analyzed using a fusion construct of the 1,766 bp AtMTM1 promoter and the GUS (β-glucuronidase) reporter gene. The screen identified GUS reporter gene expression in the developing cotyledons, leaves, roots, stems, and flowers but not in the siliques. Our results suggest that AtMTM1 encodes a mitochondrial protein that may be playing an important role in activation of MnSOD1 in Arabidopsis.


AtMTM1 Mitochondria Superoxide dismutase Activation Manganese 



Copper chaperone for superoxide dismutase


Green fluorescence protein




Manganese trafficking factor for mitochondrial SOD2


Nitroblue tetrazolium


Natural resistance associated macrophage protein


Reactive oxygen species


Superoxide dismutase


Yeast extract and peptone-based medium with 2% galactose


  1. Archibald F (2003) Oxygen toxicity and the health and survival of eukaryote cells: a new piece is added to the puzzle. Proc Natl Acad Sci USA 100:10141–10143PubMedCrossRefGoogle Scholar
  2. Aschner M, Erikson KM, Dorman DC (2005) Manganese dosimetry: species differences and implications for neurotoxicity. Crit Rev Toxicol 35:1–32PubMedCrossRefGoogle Scholar
  3. Barondeau DP, Kassmann CJ, Bruns CK, Tainer JA, Getzoff ED (2004) Nickel superoxide dismutase structure and mechanism. Biochemistry 43:8038–8047PubMedCrossRefGoogle Scholar
  4. Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem 44:276–287PubMedCrossRefGoogle Scholar
  5. Charrier B, Champion A, Henry Y, Kreis M (2002) Expression profiling of the whole Arabidopsis shaggy-like kinase multigene family by real-time reverse transcriptase-polymerase chain reaction. Plant Physiol 130:577–590PubMedCrossRefGoogle Scholar
  6. Chen XZ, Peng JB, Cohen A, Nelson H, Nelson N, Hediger MA (1999) Yeast SMF1 mediates H(+)-coupled iron uptake with concomitant uncoupled cation currents. J Biol Chem 274:35089–35094PubMedCrossRefGoogle Scholar
  7. Chu CC, Lee WC, Guo WY, Pan SM, Chen LJ, Li HM, Jinn TL (2005) A copper chaperone for superoxide dismutase that confers three types of copper/zinc superoxide dismutase activity in Arabidopsis. Plant Physiol 139:425–436PubMedCrossRefGoogle Scholar
  8. Cohen A, Nelson H, Nelson N (2000) The family of SMF metal ion transporters in yeast cells. J Biol Chem 275:33388–33394PubMedCrossRefGoogle Scholar
  9. Culotta VC, Yang M, Hall MD (2005) Manganese transport and trafficking: lessons learned from Saccharomyces cerevisiae. Eukaryot Cell 4:1159–1165PubMedCrossRefGoogle Scholar
  10. Curie C, Alonso JM, Le Jean M, Ecker JR, Briat JF (2000) Involvement of NRAMP1 from Arabidopsis thaliana in iron transport. Biochem J 347(Pt 3):749–755PubMedCrossRefGoogle Scholar
  11. Gratao PL, Polle A, Lea PJ, Azevedo RA (2005) Making the life of heavy metal-stressed plants a little easier. Funct Plant Biol 32:481–494CrossRefGoogle Scholar
  12. 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
  13. Jackson C, Dench J, Moore AL, Halliwell B, Foyer CH, Hall DO (1978) Subcellular localisation and identification of superoxide dismutase in the leaves of higher plants. Eur J Biochem 91:339–344PubMedCrossRefGoogle Scholar
  14. Jefferson RA (1989) The GUS reporter gene system. Nature 342:837–838PubMedCrossRefGoogle Scholar
  15. Kaiser BN, Moreau S, Castelli J, Thomson R, Lambert A, Bogliolo S, Puppo A, Day DA (2003) The soybean NRAMP homologue, GmDMT1, is a symbiotic divalent metal transporter capable of ferrous iron transport. Plant J 35:295–304PubMedCrossRefGoogle Scholar
  16. Lah MS, Dixon MM, Pattridge KA, Stallings WC, Fee JA, Ludwig ML (1995) Structure-function in Escherichia coli iron superoxide dismutase: comparisons with the manganese enzyme from Thermus thermophilus. Biochemistry 34:1646–1660PubMedCrossRefGoogle Scholar
  17. Leclere V, Boiron P, Blondeau R (1999) News & notes: diversity of superoxide-dismutases among clinical and soil isolates of Streptomyces species. Curr Microbiol 39:365–368PubMedCrossRefGoogle Scholar
  18. Leon S, Touraine B, Briat JF, Lobreaux S (2002) The AtNFS2 gene from Arabidopsis thaliana encodes a NifS-like plastidial cysteine desulphurase. Biochem J 366:557–564PubMedCrossRefGoogle Scholar
  19. Luk E, Carroll M, Baker M, Culotta VC (2003) Manganese activation of superoxide dismutase 2 in Saccharomyces cerevisiae requires MTM1, a member of the mitochondrial carrier family. Proc Natl Acad Sci USA 100:10353–10357PubMedCrossRefGoogle Scholar
  20. Luk E, Yang M, Jensen LT, Bourbonnais Y, Culotta VC (2005) Manganese activation of superoxide dismutase 2 in the mitochondria of Saccharomyces cerevisiae. J Biol Chem 280:22715–22720PubMedCrossRefGoogle Scholar
  21. Luk EE, Culotta VC (2001) Manganese superoxide dismutase in Saccharomyces cerevisiae acquires its metal co-factor through a pathway involving the Nramp metal transporter, Smf2p. J Biol Chem 276:47556–47562PubMedCrossRefGoogle Scholar
  22. McCord JM, Fridovich I (1969) Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein). J Biol Chem 244:6049–6055PubMedGoogle Scholar
  23. Millar AH, Heazlewood JL (2003) Genomic and proteomic analysis of mitochondrial carrier proteins in Arabidopsis. Plant Physiol 131:443–453PubMedCrossRefGoogle Scholar
  24. Okado-Matsumoto A, Fridovich I (2001) Subcellular distribution of superoxide dismutases (SOD) in rat liver: Cu, Zn-SOD in mitochondria. J Biol Chem 276:38388–38393PubMedCrossRefGoogle Scholar
  25. Outten CE, Culotta VC (2003) A novel NADH kinase is the mitochondrial source of NADPH in Saccharomyces cerevisiae. Embo J 22:2015–2024PubMedCrossRefGoogle Scholar
  26. Portnoy ME, Liu XF, Culotta VC (2000) Saccharomyces cerevisiae expresses three functionally distinct homologues of the nramp family of metal transporters. Mol Cell Biol 20:7893–7902PubMedCrossRefGoogle Scholar
  27. Ravindranath SD, Fridovich I (1975) Isolation and characterization of a manganese-containing superoxide dismutase from yeast. J Biol Chem 250:6107–6112PubMedGoogle Scholar
  28. Robinson AJ, Kunji ER (2006) Mitochondrial carriers in the cytoplasmic state have a common substrate binding site. Proc Natl Acad Sci USA 103:2617–2622PubMedCrossRefGoogle Scholar
  29. Thomine S, Wang R, Ward JM, Crawford NM, Schroeder JI (2000) Cadmium and iron transport by members of a plant metal transporter family in Arabidopsis with homology to Nramp genes. Proc Natl Acad Sci USA 97:4991–4996PubMedCrossRefGoogle Scholar
  30. Yang M, Cobine PA, Molik S, Naranuntarat A, Lill R, Winge DR, Culotta VC (2006) The effects of mitochondrial iron homeostasis on cofactor specificity of superoxide dismutase 2. Embo J 25:1775–1783PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Zhao Su
    • 1
  • Mao-Feng Chai
    • 1
  • Ping-Li Lu
    • 1
  • Rui An
    • 1
  • Jia Chen
    • 1
  • Xue-Chen Wang
    • 1
  1. 1.State Key Laboratory of Plant Physiology and Biochemistry, College of Biological SciencesChina Agricultural UniversityBeijingChina

Personalised recommendations