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Planta

, Volume 249, Issue 2, pp 417–429 | Cite as

The interaction between AtMT2b and AtVDAC3 affects the mitochondrial membrane potential and reactive oxygen species generation under NaCl stress in Arabidopsis

  • Min Zhang
  • Shenkui Liu
  • Tetsuo Takano
  • Xinxin ZhangEmail author
Original Article
  • 165 Downloads

Abstract

Main conclusion

AtMT2b interacts with AtVDAC3 in mitochondria in Arabidopsis. The overexpression of the AtMT2b and AtVDAC3 T-DNA insertion mutant confers tolerance to NaCl stress in Arabidopsis. Both AtMT2b and AtVDAC3 are involved in the regulation of the mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) under NaCl stress.

Metallothioneins (MTs) are small, cysteine rich, metal-binding proteins that perform multiple functions, such as heavy metal detoxification and reactive oxygen species (ROS) scavenging. MTs have been reported to be involved in mitochondrial function in mammals. However, whether a direct relationship exists between MTs and mitochondrial proteins remains unclear. In the present study, we used yeast two-hybrid and bimolecular fluorescence complementation assays to demonstrate that AtMT2b, which is a type 2 MT in Arabidopsis, interacts with the outer mitochondrial membrane voltage-dependent anion channel AtVDAC3. AtMT2b bound AtVDAC3, leading to its co-localization in mitochondria. AtMT2b transgenic seedlings exhibited increased tolerance to salt stress, and the atvdac3 mutant showed a similar phenotype. The mitochondrial membrane potential (MMP) was maintained, and ROS generation was reduced following AtMT2b overexpression and AtVDAC3 knockout under NaCl stress. Both AtMT2b and AtVDAC3 were shown to be involved in MMP regulation and ROS production under NaCl stress but showed opposite effects. We conclude that AtMT2b might negatively interact with AtVDAC3 in mitochondria, and both proteins are involved in the regulation of MMP and ROS under NaCl stress.

Keywords

Gene expression Metallothionein Mitochondria Protein interaction Salt tolerance Voltage-dependent anion channel 

Abbreviations

BiFC

Bimolecular fluorescence complementation

MMP

Mitochondrial membrane potential

MTs

Metallothioneins

OMM

Outer mitochondrial membrane

ROS

Reactive oxygen species

VDACs

Voltage-dependent anion channels

Y2H

Yeast two-hybrid

Notes

Acknowledgements

This work was supported by funding from the Changjiang Scholars and Innovative Research Team in University (PCSIRT) (IRT_17R99) to Shenkui Liu and a grant from the Fundamental Research Funds for the Central Universities (2572014DA06) to Xinxin Zhang. We thank professor Lixin Li for providing the BY2 cells.

Compliance with ethical standards

Conflict of interest

The authors have no conflicts of interest to declare.

Supplementary material

425_2018_3010_MOESM1_ESM.docx (3.3 mb)
Supplementary material 1 (DOCX 3404 kb)

References

  1. Al Bitar F, Roosens N, Smeyers M, Vauterin M, Van Boxtel J, Jacobs M, Homble F (2003) Sequence analysis, transcriptional and posttranscriptional regulation of the rice vdac family. Biochim Biophys Acta 1625:43–51CrossRefGoogle Scholar
  2. Batandier C, Leverve X, Fontaine E (2004) Opening of the mitochondrial permeability transition pore induces reactive oxygen species production at the level of the respiratory chain complex I. J Biol Chem 279:17197–17204CrossRefGoogle Scholar
  3. Cai L, Wang Y, Zhou G, Chen T, Song Y, Li X, Kang YJ (2006) Attenuation by metallothionein of early cardiac cell death via suppression of mitochondrial oxidative stress results in a prevention of diabetic cardiomyopathy. J Am Coll Cardiol 48:1688–1697CrossRefGoogle Scholar
  4. Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743CrossRefGoogle Scholar
  5. Costello LC, Guan Z, Franklin RB, Feng P (2004) Metallothionein can function as a chaperone for zinc uptake transport into prostate and liver mitochondria. J Inorg Biochem 98:664–666CrossRefGoogle Scholar
  6. Desai MK, Mishra RN, Verma D, Nair S, Sopory SK, Reddy MK (2006) Structural and functional analysis of a salt stress inducible gene encoding voltage dependent anion channel (VDAC) from pearl millet (Pennisetum glaucum). Plant Physiol Biochem 44:483–493CrossRefGoogle Scholar
  7. Dong F, Li Q, Sreejayan N, Nunn JM, Ren J (2007) Metallothionein prevents high-fat diet induced cardiac contractile dysfunction: role of peroxisome proliferator activated receptor coactivator 1 and mitochondrial biogenesis. Diabetes 56:2201–2212CrossRefGoogle Scholar
  8. Feng W, Cai J, Pierce WM, Franklin RB, Maret W, Benz FW, Kang YJ (2005) Metallothionein transfers zinc to mitochondrial aconitase through a direct interaction in mouse hearts. Biochem Biophys Res Commun 332:853–858CrossRefGoogle Scholar
  9. Fu Z, Guo J, Jing L, Li R, Zhang T, Peng S (2010) Enhanced toxicity and ROS generation by doxorubicin in primary cultures of cardiomyocytes from neonatal metallothionein-I/II null mice. Toxicol In Vitro 24:1584–1591CrossRefGoogle Scholar
  10. Futakawa N, Kondoh M, Ueda S, Higashimoto M, Takiguchi M, Suzuki S, Sato M (2006) Involvement of oxidative stress in the synthesis of metallothionein induced by mitochondrial inhibitors. Biol Pharm Bull 29:2016–2020CrossRefGoogle Scholar
  11. Godbole A, Mitra R, Dubey AK, Reddy PS, Mathew MK (2011) Bacterial expression, purification and characterization of a rice voltage-dependent, anion-selective channel isoform, OsVDAC4. J Membr Biol 244:67–80CrossRefGoogle Scholar
  12. Godbole A, Dubey AK, Reddy PS, Udayakumar M, Mathew MK (2013) Mitochondrial VDAC and hexokinase together modulate plant programmed cell death. Protoplasma 250:875–884CrossRefGoogle Scholar
  13. Han D, Antunes F, Canali R, Rettori D, Cadenas E (2002) Voltage-dependent anion channels control the release of the superoxide anion from mitochondria to cytosol. J Biol Chem 278:5557–5563CrossRefGoogle Scholar
  14. Heazlewood JL, Pan X, Chen Z, Yang X, Liu G (2014) Arabidopsis voltage-dependent anion channel 1 (AtVDAC1) is required for female development and maintenance of mitochondrial functions related to energy-transaction. PLoS One 9(9):e106941CrossRefGoogle Scholar
  15. Kondoh M, Inoue Y, Atagi S, Futakawa N, Higashimoto M, Sato M (2001) Specific induction of metallothionein synthesis by mitochondrial oxidative stress. Life Sci 69:2137–2146CrossRefGoogle Scholar
  16. Kusano T, Tateda C, Berberich T, Takahashi Y (2009) Voltage-dependent anion channels: their roles in plant defense and cell death. Plant Cell Rep 28:1301–1308CrossRefGoogle Scholar
  17. Lacomme C, Roby D (1999) Identification of new early markers of the hypersensitive response in Arabidopsis thaliana 1. FEBS Lett 459:149–153CrossRefGoogle Scholar
  18. Lee SM, Hoang MHT, Han HJ, Kim HS, Lee K, Kim KE, Kim DH, Lee SY, Chung WS (2009) Pathogen inducible voltage-dependent anion channel (AtVDAC) isoforms are localized to mitochondria membrane in Arabidopsis. Mol Cells 27:321–327CrossRefGoogle Scholar
  19. Li Z-Y, Xu Z-S, He G-Y, Yang G-X, Chen M, Li L-C, Ma Y (2013) The voltage-dependent anion channel 1 (AtVDAC1) negatively regulates plant cold responses during germination and seedling development in Arabidopsis and interacts with calcium sensor CBL1. Int J Mol Sci 14:701–713CrossRefGoogle Scholar
  20. Lindeque JZ, Levanets O, Louw R, van der Westhuizen FH (2010) The involvement of metallothioneins in mitochondrial function and disease. Curr Protein Pept Sci 11:292–309CrossRefGoogle Scholar
  21. Martel C, Allouche M, Esposti DD, Fanelli E, Boursier C, Henry C, Chopineau J, Calamita G, Kroemer G, Lemoine A, Brenner C (2013) Glycogen synthase kinase 3-mediated voltage-dependent anion channel phosphorylation controls outer mitochondrial membrane permeability during lipid accumulation. Hepatology 57:93–102CrossRefGoogle Scholar
  22. Miyayama T, Arai Y, Suzuki N, Hirano S (2013) Mitochondrial electron transport is inhibited by disappearance of metallothionein in human bronchial epithelial cells following exposure to silver nitrate. Toxicology 305:20–29CrossRefGoogle Scholar
  23. Moltó E, Bonzón-Kulichenko E, Gallardo N, Andrés A (2007) MTPA: a crustacean metallothionein that affects hepatopancreatic mitochondrial functions. Arch Biochem Biophys 467:31–40CrossRefGoogle Scholar
  24. Robert N, d’Erfurth I, Marmagne A, Erhardt M, Allot M, Boivin K, Gissot L, Monachello D, Michaud M, Duchêne A-M, Barbier-Brygoo H, Maréchal-Drouard L, Ephritikhine G, Filleur S (2012) Voltage-dependent-anion-channels (VDACs) in Arabidopsis have a dual localization in the cell but show a distinct role in mitochondria. Plant Mol Biol 78:431–446CrossRefGoogle Scholar
  25. Rostovtseva TK, Bezrukov SM (1998) ATP transport through a single mitochondrial channel, VDAC, studied by current fluctuation analysis. Biophys J 74:2365–2373CrossRefGoogle Scholar
  26. Rostovtseva TK, Bezrukov SM (2008) VDAC regulation: role of cytosolic proteins and mitochondrial lipids. J Bioenerg Biomembr 40:163–170CrossRefGoogle Scholar
  27. Rostovtseva T, Colombini M (1997) VDAC channels mediate and gate the flow of ATP: implications for the regulation of mitochondrial function. Biophys J 72:1954–1962CrossRefGoogle Scholar
  28. Sampson MJ, Lovell RS, Craigen WJ (1997) The murine voltage-dependent anion channel gene family: conserved structure and function. J Biol Chem 272:18966–18973CrossRefGoogle Scholar
  29. Sanchez Ferrer A, Santema JS, Hilhorst R, Visser AJ (1990) Fluorescence detection of enzymatically formed hydrogen peroxide in aqueous solution and in reversed micelles. Anal Biochem 187:129–132CrossRefGoogle Scholar
  30. Simpkins C, Balderman S, Mensah E (1998a) Mitochondrial oxygen consumption is synergistically inhibited by metallothionein and calcium. J Surg Res 80:16–21CrossRefGoogle Scholar
  31. Simpkins C, Lloyd T, Li S, Balderman S (1998b) Metallothionein-induced increase in mitochondrial inner membrane permeability. J Surg Res 75:30–34CrossRefGoogle Scholar
  32. Takahashi Y, Tateda C (2013) The functions of voltage-dependent anion channels in plants. Apoptosis 18:917–924CrossRefGoogle Scholar
  33. Tan W, Colombini M (2007) VDAC closure increases calcium ion flux. BBA Biomembr 1768:2510–2515CrossRefGoogle Scholar
  34. Tang W, Shaikh ZA (2001) Renal cortical mitochondrial dysfunction upon cadmium metallothionein administration to sprague-dawley rats. J Toxicol Environ Health Part A 63:221–235CrossRefGoogle Scholar
  35. Tateda C, Yamashita K, Takahashi F, Kusano T, Takahashi Y (2008) Plant voltage-dependent anion channels are involved in host defense against Pseudomonas cichorii and in Bax-induced cell death. Plant Cell Rep 28:41–51CrossRefGoogle Scholar
  36. Tateda C, Watanabe K, Kusano T, Takahashi Y (2011) Molecular and genetic characterization of the gene family encoding the voltage-dependent anion channel in Arabidopsis. J Exp Bot 62:4773–4785CrossRefGoogle Scholar
  37. Tsugama D, Liu S, Takano T (2012) A putative myristoylated 2C-type protein phosphatase, PP2C74, interacts with SnRK1 in Arabidopsis. FEBS Lett 586:693–698CrossRefGoogle Scholar
  38. Wandrey M, Trevaskis B, Brewin N, Udvardi MK (2004) Molecular and cell biology of a family of voltage-dependent anion channel porins in Lotus japonicus. Plant Physiol 134:182–193CrossRefGoogle Scholar
  39. Wang GW, Klein JB, Kang YJ (2001) Metallothionein inhibits doxorubicin-induced mitochondrial cytochrome c release and caspase-3 activation in cardiomyocytes. J Pharmacol Exp Ther 298:461–468Google Scholar
  40. Xue T, Li X, Zhu W, Wu C, Yang G, Zheng C (2009) Cotton metallothionein GhMT3a, a reactive oxygen species scavenger, increased tolerance against abiotic stress in transgenic tobacco and yeast. J Exp Bot 60:339–349CrossRefGoogle Scholar
  41. Yan J, He H, Tong S, Zhang W, Li X, Yang Y (2009) Voltage-dependent anion channel 2 of Arabidopsis thaliana (AtVDAC2) is involved in ABA-mediated early seedling development. Int J Mol Sci 10:2476–2486CrossRefGoogle Scholar
  42. Yang XY, Chen ZW, Xu T, Qu Z, Pan XD, Qin XH, Ren DT, Liu GQ (2011) Arabidopsis kinesin KP1 specifically interacts with VDAC3, a mitochondrial protein, and regulates respiration during seed germination at low temperature. Plant Cell 23:1093–1106CrossRefGoogle Scholar
  43. Ye B, Maret W, Vallee BL (2001) Zinc metallothionein imported into liver mitochondria modulates respiration. Proc Natl Acad Sci USA 98:2317–2322CrossRefGoogle Scholar
  44. Yi J, Moon S, Lee YS, Zhu L, Liang W, Zhang D, Jung KH, An G (2016) Defective tapetum cell death 1 (DTC1) regulates ROS levels by binding to metallothionein during tapetum degeneration. Plant Physiol 170:1611–1623Google Scholar
  45. Yoo SD, Cho YH, Sheen J (2007) Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis. Nat Protoc 2:1565–1572CrossRefGoogle Scholar
  46. Zalewska M, Trefon J, Milnerowicz H (2014) The role of metallothionein interactions with other proteins. Proteomics 14:1343–1356CrossRefGoogle Scholar
  47. Zhang M, Takano T, Liu S, Zhang X (2015) Arabidopsis mitochondrial voltage-dependent anion channel 3 (AtVDAC3) protein interacts with thioredoxin m2. FEBS Lett 589:1207–1213CrossRefGoogle Scholar
  48. Zheng Y, Shi Y, Tian C, Jiang C, Jin H, Chen J, Almasan A, Tang H, Chen Q (2004) Essential role of the voltage-dependent anion channel (VDAC) in mitochondrial permeability transition pore opening and cytochrome c release induced by arsenic trioxide. Oncogene 23:1239–1247CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Key Laboratory of Saline-alkali Vegetation Ecology Restoration (SAVER), Ministry of Education, Alkali Soil Natural Environmental Science Center (ASNESC)Northeast Forestry UniversityHarbinChina
  2. 2.School of MedicineHe UniversityShenyangChina
  3. 3.State Key Laboratory of Subtropical SilvicultureZhejiang A & F UniversityHangzhouChina
  4. 4.Asian Natural Environment Science Center (ANESC)The University of TokyoTokyoJapan

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