Biologia plantarum 55:173-177, 2011 | DOI: 10.1007/s10535-011-0025-7

Subcellular localization of rice hexokinase (OsHXK) family members in the mesophyll protoplasts of tobacco

W. Cheng1, H. Zhang1, X. Zhou1, H. Liu1, Y. Liu1, J. Li1, S. Han1, Y. Wang1,*
1 Laboratory of Plant Development Physiology and Molecular Biology, College of Life Sciences, Beijing Normal University, Beijing, P.R. China

Hexokinase (HXK, EC 2.7.1.1) plays an important role in the metabolism and glucose signalling. To examine the characteristics of HXK gene family in rice, the subcellular localizations of ten hexokinases (OsHXK1 - OsHXK10) were determined using OsHXK::GFP fusion proteins in tobacco mesophyll protoplasts. As was previously demonstrated, OsHXK4 was detected in the chloroplast stroma, OsHXK5 and OsHXK6 in the mitochondria, and OsHXK7 and OsHXK10 in the cytoplasm. In the present study, OsHXKs were clearly divided into three types (A, B, C) based on their N-terminal sequences. The new type-C HXKs in plants, OsHXK1, OsHXK7 and OsHXK8, which lack the plastidic transit peptide and the membrane anchor domain, were detected not only in the cytoplasm but also in the nucleus. The type-B HXKs, OsHXK2, OsHXK3, OsHXK9 and OsHXK10, which contained a membrane anchor domain, were distinctly localized in the mitochondria. These results suggest that OsHXKs localized in different cell compartments may be involved in the glucose signalling-related gene expression during growth and development of rice.

Keywords: GFP; hexose; Nicotiana tabacum; Oryza sativa

Received: April 17, 2009; Accepted: December 20, 2009; Published: March 1, 2011Show citation

ACS AIP APA ASA Harvard Chicago IEEE ISO690 MLA NLM Turabian Vancouver
Cheng, W., Zhang, H., Zhou, X., Liu, H., Liu, Y., Li, J., Han, S., & Wang, Y. (2011). Subcellular localization of rice hexokinase (OsHXK) family members in the mesophyll protoplasts of tobacco. Biologia plantarum55(1), 173-177. doi: 10.1007/s10535-011-0025-7.
Download citation

References

  1. Balasubramanian, R., Karve, A., Kandasamy, M., Meagher, R.B., Moore, B.: A role for F-actin in hexokinase-mediated glucose signalling. - Plant Physiol. 145: 1423-1434, 2007. Go to original source...
  2. Cárdenas, M.L., Cornish-Bowden, A., Ureta, T.: Evolution and regulatory role of the hexokinases. - Biochim. biophys. Acta 1401: 242-264, 1998. Go to original source...
  3. Cho, J.I., Ryoo, N., Eom, J.S., Lee, D.W., Kim, H.B., Jeong, S.W., Lee, Y.H., Kwon, Y.K., Cho, M.H., Bhoo, S.H., Hahn, T.R., Park, Y.I., Hwang, I., Sheen, J., Jeon, J.S.: Role of the rice hexokinases OsHXK5 and OsHXK6 as glucose sensors. - Plant Physiol. 149: 745-759, 2009. Go to original source...
  4. Cho, J.I., Ryoo, N., Ko, S., Lee, S.K., Lee, J., Jung, K.H., Lee, Y.H., Bhoo, S.H., Winderickx, J., An, G., Hahn, T.R., Jeon, J.S.: Structure, expression, and functional analysis of the hexokinase gene family in rice (Oryza sativa L.). - Planta 224: 598-611, 2006a. Go to original source...
  5. Cho, Y.H., Yoo, S.D., Sheen, J.: Regulatory functions of nuclear hexokinase1 complex in glucose signaling. - Cell 127: 579-589, 2006b. Go to original source...
  6. Claeyssen, E., Rivoal, J.: Isozymes of plant hexokinase: occurrence, properties and functions. - Phytochemistry 68: 709-731, 2007. Go to original source...
  7. Da Silva, W.S., Rezende, G.L., Galina, A.: Subcellular distribution and kinetic properties of cytosolic and non-cytosolic hexokinases in maize seedling roots: implications for hexose phosphorylation. - J. exp. Bot. 52: 1191-1201, 2001. Go to original source...
  8. Damari-Weissler, H., Kandel-Kfir, M., Gidoni, D., Mett, A., Belausov, E., Granot, D.: Evidence for intracellular spatial separation of hexokinases and fructokinases in tomato plants. - Planta 224: 1495-1502, 2006. Go to original source...
  9. Damari-Weissler, H., Ginzburg, A., Gidoni, D., Mett, A., Krassovskaya, I., Weber, A.P., Belausov, E., Granot, D.: Spinach SoHXK1 is a mitochondria-associated hexokinase. - Planta 226: 1053-1058, 2007. Go to original source...
  10. Emanuelsson, O., Nielsen, H., Brunak, S., Von Heijine, G.: Predicting subcellular localization of proteins based on their N-terminal amino acid sequence. - J. mol. Biol. 300: 1005-1016, 2000. Go to original source...
  11. Galina, A., Reis, M., Albuquerque, M.C., Puyou, A.G., Puyou, M.T., de Meis, L.: Different properties of the mitochondrial and cytosolic hexokinases in maize roots. - Biochem. J. 309(Pt 1):105-112, 1995. Go to original source...
  12. Giege, P., Heazlewood, J.L., Roessner-Tunali, U., Millar, A.H., Fernie, A.R., Leaver, C.J., Sweetlove, L.J.: Enzymes of glycolysis are functionally associated with the mitochondrion in Arabidopsis cells. - Plant Cell 15: 2140-2151, 2003. Go to original source...
  13. Giese, J.O., Herbers, K., Hoffmann, M., Klosgen, R.B., Sonnewald, U.: Isolation and functional characterization of a novel plastidic hexokinase from Nicotiana tabacum. - FEBS Lett. 579: 827-831, 2005. Go to original source...
  14. Higuchi, R.: Recombinant PCR. - In: Innis, M.A., Gelfand, D.H., Sninsky, J.J., White, T.J. (ed.): PCR Protocols: A Guide to Methods and Applications. Pp. 177-182. Academic Press, New York 1990. Go to original source...
  15. Kandel-Kfir, M., Damari-Weissler, H., German, M.A., Gidoni, D., Mett, A., Belausov, E., Petreikov, M., Adir, N., Granot, D.: Two newly identified membrane-associated and plastidic tomato HXKs: characteristics, predicted structure and intracellular localization. - Planta 224: 1341-1352, 2006. Go to original source...
  16. Karve, A., Rauh, B.L., Xia, X., Kandasamy, M., Meagher, R.B., Sheen, J., Moore, B.D.: Expression and evolutionary features of the hexokinase gene family in Arabidopsis. - Planta 228: 411-425, 2008. Go to original source...
  17. Kim, M., Lim, J.H., Ahn, C.S., Park, K., Kim, G.T., Kim, W.T., Pai, H.S.: Mitochondria-associated hexokinases play a role in the control of programmed cell death in Nicotiana benthamiana. - Plant Cell 18: 2341-2355, 2006. Go to original source...
  18. Koch, K.: Sucrose metabolism: regulatory mechanisms and pivotal roles in sugar sensing and plant development. - Curr.t Opin. Plant Biol. 7: 235-246, 2004. Go to original source...
  19. Moore, B., Zhou, L., Rolland, F., Hall, Q., Cheng, W.H., Liu, Y.X., Hwang, I., Jones, T., Sheen, J.: Role of the Arabidopsis glucose sensor HXK1 in nutrient, light, and hormonal signalling. - Science 300: 332-336, 2003. Go to original source...
  20. Nielsen, H., Engelbrecht, J., Brunak, S., von Heijne, G.: Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites. - Protein Eng. 10: 1-6, 1997. Go to original source...
  21. Olsson, T., Thelander, M., Ronne, H.: A novel type of chloroplast stromal hexokinase is the major glucose-phosphorylating enzyme in the moss Physcomitrella patens. - J. biol. Chem. 278: 44439-44447, 2003. Go to original source...
  22. Randez-Gil, F., Herrero, P., Sanz, P., Prieto, J.A., Moreno, F.: Hexokinase PII has a double cytosolic-nuclear localisation in Saccharomyces cerevisiae. - FEBS Lett. 425: 475-478, 1998. Go to original source...
  23. Rezende, G.L., Logullo, C., Meyer, L., Machado, L.B., Oliveira-Carvalho, A.L., Zingali, R.B., Cifuentes, D., Galina, A.: Partial purification of tightly bound mitochondrial hexokinase from maize (Zea mays L.) root membranes. - Braz. J. Med. biol. Res. 39: 1159-1169, 2006. Go to original source...
  24. Rodríguez, A., De La Cera, T., Herrero, P., Moreno, F.: The hexokinase 2 protein regulates the expression of the GLK1, HXK1 and HXK2 genes of Saccharomyces cerevisiae. - Biochem. J. 355: 625-631, 2001. Go to original source...
  25. Šindelář, L., Šindelářová, M.: Hexokinases of tobacco leaves: changes in the cytosolic and non-cytosolic isozyme complexes induced by tobacco mosaic virus infection. - Biol. Plant. 47: 413-419, 2003.
  26. Šindelářová, M., Šindelář, L.: Hexokinases of tobacco leaves: subcellular localization and characterization. - Biol. Plant. 30: 275-284, 1988.
  27. Singh, K.K., Chen, C., Epstein, D.K., Gibbs, M.: 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: 587-593, 1993. Go to original source...
  28. Xu, F.Q., Li, X.R., Ruan, Y.L.: RNAi-mediated suppression of hexokinase gene OsHXK10 in rice leads to non-dehiscent anther and reduction of pollen germination. - Plant Sci. 175: 674-684, 2008. Go to original source...
  29. Yoo, S.D., Cho, Y.H., Sheen, J.: Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis. - Nat. Protocol. 2: 1565-1572, 2007.