Advertisement

Planta

, Volume 226, Issue 4, pp 1053–1058 | Cite as

Spinach SoHXK1 is a mitochondria-associated hexokinase

  • Hila Damari-Weissler
  • Alexandra Ginzburg
  • David Gidoni
  • Anahit Mett
  • Inga Krassovskaya
  • Andreas P. M. Weber
  • Eddy Belausov
  • David GranotEmail author
Rapid Communication

Abstract

Hexokinase, a hexose-phosphorylating enzyme, has emerged as a central enzyme in sugar-sensing processes. A few HXK isozymes have been identified in various plant species. These isozymes have been classified into two major groups; plastidic (type A) isozymes located in the plastid stroma and those containing a membrane anchor domain (type B) located mainly adjacent to the mitochondria, but also found in the nucleus. Of all the hexokinases that have been characterized to date, the only exception to this rule is a spinach type B HXK (SoHXK1) that, by means of subcellular fractionation, has been localized to the outer membrane of plastids. However, SoHXK1 has a membrane anchor domain that is almost identical to that of the other type B HXKs. To determine the localization of SoHXK1 enzyme by other means, we expressed SoHXK1::GFP fusion protein in tobacco and Arabidopsis protoplasts and compared its localization with that of the Arabidopsis AtHXK1::GFP fusion protein that shares a similar N-terminal membrane anchor domain. SoHXK1::GFP is localized adjacent to the mitochondria, similar to AtHXK1::GFP and all other previously examined type B HXKs. Proteomic analysis had previously identified AtHXK1 on the outside of the mitochondrial membrane. We, therefore, suggest that SoHXK1 enzyme is located adjacent to the mitochondria like the other type B HXKs that share the same N-terminal membrane anchor domain.

Keywords

GFP Hexokinase Hexose phosphorylation Intracellular localization 

Abbreviations

GFP

Green fluorescent protein

HXK

Hexokinase

Notes

Acknowledgments

This research was supported by research grant No. 890/06 from The Israel Science Foundation, research grants No. IS-3397-06 and No. CA-9100-06 from BARD, the United States—Israel Binational Agricultural and Development Fund, and a grant from the US Department of Energy to A.P.M.W. (DE-FG02-04ER15562).

References

  1. Bauer M, Dietrich C, Nowak K, Sierralta WD, Papenbrock J (2004) Intracellular localization of Arabidopsis sulfurtransferases. Plant Physiol 135:916–926PubMedCrossRefGoogle Scholar
  2. Cho JI, Ryoo N, Ko S, Lee SK, Lee J, Jung KH, Lee YH, Bhoo SH, Winderickx J, An G, Hahn TR, Jeon JS (2006a) Structure, expression, and functional analysis of the hexokinase gene family in rice (Oryza sativa L.). Planta 224:598–611PubMedCrossRefGoogle Scholar
  3. Cho YH, Yoo SD, Sheen J (2006b) Regulatory functions of nuclear hexokinase1 complex in glucose signaling. Cell 127:579–589PubMedCrossRefGoogle Scholar
  4. Cosio E, Bustamante E (1984) Subcellular localization of hexokinase in pea leaves. Evidence for the predominance of a mitochondrially bound form. J Biol Chem 259:7688–7692PubMedGoogle Scholar
  5. Curtis MD, Grossniklaus U (2003) A gateway cloning vector set for high-throughput functional analysis of genes in planta. Plant Physiol 133:462–469PubMedCrossRefGoogle Scholar
  6. Dai N, Schaffer A, Petreikov M, Granot D (1995) Cloning of Arabidopsis thaliana hexokinase cDNA by complementation of yeast cells. Plant Physiol 108:879–880PubMedCrossRefGoogle Scholar
  7. Dai N, Kandel M, Petreikov M, Levine I, Ricard B, Rothan C, Schaffer AA, Granot D (2002) The tomato hexokinase LeHXK1: cloning, mapping, expression pattern and phylogenetic relationships. Plant Sci 163:581–590CrossRefGoogle Scholar
  8. Damari-Weissler H, Kandel-Kfir M, Gidoni D, Mett A, Belausov E, Granot D (2006) Evidence for intracellular spatial separation of hexokinases and fructokinases in tomato plants. Planta 224:1495–1502PubMedCrossRefGoogle Scholar
  9. da-Silva WS, Rezende GL, Galina A (2001) Subcellular distribution and kinetic properties of cytosolic and non-cytosolic hexokinases in maize seedling roots: implications for hexose phosphorylation. J Exp Bot 52:1191–1201PubMedCrossRefGoogle Scholar
  10. Deuschle K, Chaudhuri B, Okumoto S, Lager I, Lalonde S, Frommer WB (2006) Rapid metabolism of glucose detected with FRET glucose nanosensors in epidermal cells and intact roots of Arabidopsis RNA-silencing mutants. Plant Cell 18:2314–2325PubMedCrossRefGoogle Scholar
  11. Draper J, Scott R, Hamil J (1988) Transformation of dicotiledonous plant cells using the Ti plasmid of A. tumefaciens and the Ri plasmid of A. rhizogenes. Blackwell Scientific Publications, OxfordGoogle Scholar
  12. Dry IB, Nash D, Wiskish TJ (1983) The mitochondrial localization of hexokinase in pea leaves. Planta 158:152–156CrossRefGoogle Scholar
  13. Emanuelsson O, Nielsen H, Brunak S, von Heijne G (2000) Predicting subcellular localization of proteins based on their N-terminal amino acid sequence. J Mol Biol 300:1005–1016PubMedCrossRefGoogle Scholar
  14. Ferro M, Salvi D, Brugiere S, Miras S, Kowalski S, Louwagie M, Garin J, Joyard J, Rolland N (2003) Proteomics of the chloroplast envelope membranes from Arabidopsis thaliana. Mol Cell Proteomics 2:325–345PubMedGoogle Scholar
  15. Fromm M, Taylor LP, Walbot V (1985) Expression of genes transferred into monocot and dicot plant cells by electroporation. Proc Natl Acad Sci USA 82:5824–5828PubMedCrossRefGoogle Scholar
  16. Galina A, Reis M, Albuquerque MC, Puyou AG, Puyou MT, de Meis L (1995) Different properties of the mitochondrial and cytosolic hexokinases in maize roots. Biochem J 309:105–112PubMedGoogle Scholar
  17. German MA, Asher I, Petreikov M, Dai N, Schaffer AA, Granot D (2004) Cloning, expression and characterization of LeFRK3, the fourth tomato (Lycopersicon esculentum Mill.) gene encoding fructokinase. Plant Sci 166:285–291CrossRefGoogle Scholar
  18. Giege P, Heazlewood JL, Roessner-Tunali U, Millar AH, Fernie AR, Leaver CJ, Sweetlove LJ (2003) Enzymes of glycolysis are functionally associated with the mitochondrion in Arabidopsis cells. Plant Cell 15:2140–2151PubMedCrossRefGoogle Scholar
  19. Giese JO, Herbers K, Hoffmann M, Klosgen RB, Sonnewald U (2005) Isolation and functional characterization of a novel plastidic hexokinase from Nicotiana tabacum. FEBS Lett 579:827–831PubMedCrossRefGoogle Scholar
  20. Gleave AP (1992) A versatile binary vector system with a T-DNA organisational structure conducive to efficient integration of cloned DNA into the plant genome. Plant Mol Biol 20:1203–1207PubMedCrossRefGoogle Scholar
  21. Granot D (2007) Role of tomato hexose kinases. Funct Plant Biol 34 (in press)Google Scholar
  22. Heazlewood JL, Tonti-Filippini JS, Gout AM, Day DA, Whelan J, Millar AH (2004) Experimental analysis of the Arabidopsis mitochondrial proteome highlights signaling and regulatory components, provides assessment of targeting prediction programs, and indicates plant-specific mitochondrial proteins. Plant Cell 16:241–256PubMedCrossRefGoogle Scholar
  23. Kanayama Y, Dai N, Granot D, Petreikov M, Schaffer A, Bennett AB (1997) Divergent fructokinase genes are differentially expressed in tomato. Plant Physiol 113:1379–1384PubMedCrossRefGoogle Scholar
  24. Kanayama Y, Granot D, Dai N, Petreikov M, Schaffer A, Powell A, Bennett AB (1998) Tomato fructokinases exhibit differential expression and substrate regulation. Plant Physiol 117:85–90PubMedCrossRefGoogle Scholar
  25. Kandel-Kfir M, Damari-Weissler H, German MA, Gidoni D, Mett A, Belausov E, Petreikov M, Adir N, Granot D (2006) Two newly identified membrane-associated and plastidic tomato HXKs: characteristics, predicted structure and intracellular localization. Planta 224:1341–1352PubMedCrossRefGoogle Scholar
  26. Kim M, Lim JH, Ahn CS, Park K, Kim GT, Kim WT, Pai HS (2006) Mitochondria-associated hexokinases play a role in the control of programmed cell death in Nicotiana benthamiana. Plant Cell 18:2341–2355PubMedCrossRefGoogle Scholar
  27. Menu T, Rothan C, Dai N, Petreikov M, Etienne C, Destrac-Irvine A, Schaffer A, Granot D, Ricard B (2001) Cloning and characterization of a cDNA encoding hexokinase from tomato. Plant Sci 160:209–218PubMedCrossRefGoogle Scholar
  28. Miernyk JA, Dennis DT (1983) Mitochondrial, plastid, and cytosolic isozymes of hexokinase from developing endosperm of Ricinus communis. Arch Biochem Biophys 226:458–468PubMedCrossRefGoogle Scholar
  29. Murayama S, Handa H (2007) Genes for alkaline/neutral invertase in rice: alkaline/neutral invertases are located in plant mitochondria and also in plastids. Planta 225:1193–1203 PubMedCrossRefGoogle Scholar
  30. Olsson T, Thelander M, Ronne H (2003) A novel type of chloroplast stromal hexokinase is the major glucose-phosphorylating enzyme in the moss Physcomitrella patens. J Biol Chem 278:44439–44447PubMedCrossRefGoogle Scholar
  31. Renz A, Stitt M (1993) Substrate specificity and product inhibition of different forms of fructokinases and hexokinases in developing potato tubers. Planta 190:166–175Google Scholar
  32. Rolland F, Sheen J (2005) Sugar sensing and signalling networks in plants. Biochem Soc Trans 33:269–271PubMedCrossRefGoogle Scholar
  33. Schnarrenberger C (1990) Characterization and compartmentation in green leaves, of hexokinases with different specificities for glucose, fructose, and mannose and for nucleoside triphosphates. Planta 181:249–255CrossRefGoogle Scholar
  34. Singh KK, Chen C, Epstein DK, Gibbs M (1993) Respiration of sugars in Spinach (Spinacia oleracea), Maize (Zea mays), and Chlamydomonas reinhardtii F-60 chloroplasts with emphasis on the hexose kinases. Plant Physiol 102:587–593PubMedGoogle Scholar
  35. Smith AM, Zeeman SC, Smith SM (2005) Starch degradation. Annu Rev Plant Biol 56:73–98PubMedCrossRefGoogle Scholar
  36. Subbaiah CC, Palaniappan A, Duncan K, Rhoads DM, Huber SC, Sachs MM (2006) Mitochondrial localization and putative signaling function of sucrose synthase in maize. J Biol Chem 281:15625–15635PubMedCrossRefGoogle Scholar
  37. Tanner GJ, Copeland L, Turner JF (1983) Subcellular localization of hexose kinases in pea stems: mitochondrial hexokinase Pisum sativum. Plant Physiol 72:659–663PubMedCrossRefGoogle Scholar
  38. Wiese A, Groner F, Sonnewald U, Deppner H, Lerchl J, Hebbeker U, Flugge U, Weber A (1999) Spinach hexokinase I is located in the outer envelope membrane of plastids. FEBS Lett 461:13–18PubMedCrossRefGoogle Scholar
  39. Yamamoto YT, Prata RTN, Williamson JD, Weddington M, Pharr DM (2000) Formation of a hexokinase complex is associated with changes in energy utilization in celery organs and cells. Plant Physiol 110:28–37CrossRefGoogle Scholar
  40. Zeeman SC, Smith SM, Smith AM (2007) The diurnal metabolism of leaf starch. Biochem J 401:13–28PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Hila Damari-Weissler
    • 1
  • Alexandra Ginzburg
    • 1
  • David Gidoni
    • 1
  • Anahit Mett
    • 1
  • Inga Krassovskaya
    • 2
  • Andreas P. M. Weber
    • 2
  • Eddy Belausov
    • 1
  • David Granot
    • 1
    Email author
  1. 1.Institute of Plant Sciences, Agricultural Research OrganizationThe Volcani CenterBet DaganIsrael
  2. 2.Department of Plant BiologyMichigan State UniversityEast LansingUSA

Personalised recommendations