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Hexokinases of Tobacco Leaves: Changes in the Cytosolic and Non-Cytosolic Isozyme Complexes Induced by Tobacco Mosaic Virus Infection

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Biologia Plantarum

Abstract

Changes in the cytosotic (soluble) and the non-cytosolic (particulate) isozyme composition of hexokinases and in their properties were studied by ion exchange chromatography on DEAE cellulose after the subcellular fractionation both in the healthy and the tobacco mosaic virus (TMV) infected tobacco leaves. Three main isozyme complexes were obtained: one particulate fraction (the particulate hexokinase phosphorylating both glucose and fructose, EC 2.7.1.1), and two soluble fractions (the soluble hexokinase phosphorylating both the glucose and the fructose, and the soluble fructokinase, which phosphorylates primarily fructose, EC 2.7.1.4). The total fructokinase activities were nearly twice higher than the total glucokinase activities (188.6 % of glucokinase activity in healthy plants and 181.3 % in infected plants). The total particulate glucokinase activity was increased to 120.6 % and the fructokinase to 118.9 % in TMV infected tissue when compared with healthy control. The similar pattern of activity was observed for soluble hexokinase isozymes - the sum of soluble glucokinase activity was increased to 175.4 % and of fructokinase activity to 131.2 % in TMV infected tissue. The isozymes isolated both from the healthy control and TMV-infected leaves had the similar elution profiles, displayed Michaelis-Menten kinetics, showed the identical profiles of pH optima and were Mg2+ dependent with the highest enzyme activity at equimolar Mg2+ and ATP concentration.

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References

  • Baldus, B., Kelly, G. J., Latzko, E.: Hexokinases of spinach leaves.-Phytochemistry 20: 1811–1814, 1981.

    Article  CAS  Google Scholar 

  • Clark, M.F., Adams, A.N.: Characteristics of the microplate method of enzyme-linked immunosorbent assay for the detection of plant viruses.-J. gen. Virol. 34: 473–483, 1977.

    Google Scholar 

  • Dai, N., Schaffer, A.A., Petreikov, M., Granot, D.: Arabidopsis thaliana hexokinase cDNA isolated by complementation of yeast cells.-Plant Physiol. 108: 879–880, 1995.

    Article  PubMed  CAS  Google Scholar 

  • Dai, N., Schaffer, A.A., Petreikov, M., Shahak, Y., Giller, Y., Ratner, K., Levine, A., Granot, D.: Overexpression of Arabidopsis hexokinase in tomato plants inhibits growth, reduces photosynthesis and induces rapid senescence.-Plant Cell 11: 1253–1266, 1999.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Galina, A., Da Silva, W.S.: Hexokinase activity alters sugar-nucleotide formation in maize root homogenates.-Phytochemistry 53: 29–37, 2000.

    Article  PubMed  CAS  Google Scholar 

  • 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: 105–112, 1995.

    PubMed  CAS  Google Scholar 

  • Gibson, S.I.: Plant sugar-response pathways. Part of a complex regulatory web.-Plant Physiol. 124: 1532–1539, 2000.

    Article  PubMed  CAS  Google Scholar 

  • Gooding, G.V., Hebert, T.T.: A simple technique for purification of tobacco mosaic virus in large quantities.-Phytopathology 57: 1285, 1967.

    PubMed  Google Scholar 

  • Graham, I.A., Denby, K.J., Leaver, C.J.: Carbon catabolite repression regulates glyoxylate cycle gene expression in cucumber.-Plant Cell 6: 761–772, 1994.

    Article  PubMed  CAS  Google Scholar 

  • Jang, J.-C., Leon, P., Zhou, L., Sheen, J.: Hexokinase as a sugar sensor in higher plants.-Plant Cell 9: 5–19, 1997.

    Article  PubMed  CAS  Google Scholar 

  • Jang, J.-C., Sheen, J.: Sugar sensing in higher plants.-Plant Cell 6: 1665–1679, 1994.

    Article  PubMed  CAS  Google Scholar 

  • Kapur, S.P., Gumpf, D.J., Weathers, L.G.: Some effects of metabolic changes induced in “Etrog” citron by three isolates of exocortis virus.-Phytopathology 64: 196–201, 1974.

    Article  CAS  Google Scholar 

  • Koch, K.E.: Carbohydrate-modulated gene expression in plants.-Annu. Rev. Plant Physiol. Plant mol. Biol. 47: 509–540, 1996.

    Article  PubMed  CAS  Google Scholar 

  • Kruger, N.J.: Carbohydrate synthesis and degradation. In: Dennis, D.T., Turpin, D.M. (ed.): Plant Physiology, Biochemistry and Molecular Biology. Pp. 268–292. Longman, Harlow 1990.

    Google Scholar 

  • Makovcová, O., Šindelár, L.: The effect of 2,4–dichlorophenoxyacetic acid on the metabolic utilization of free carbohydrates in cucumber mosaic virus infected cucumber plants.-Biol. Plant. 23: 465–468, 1981.

    Google Scholar 

  • Makovcová, O., Šindelár, L., HanuŠová, M.: [Sucrose metabolism in the leaves of cucumber infected with cucumber mosaic virus, as related to yields.]-Ochrana Rost. (Prague) 16: 263–269, 1980. [In Czech.]

    Google Scholar 

  • Mančal, P.: Metody Enzymové Imunoanalyzy. [Methods of Enzyme Immunoassay.]-USOL, Prague 1987. [In Czech.]

    Google Scholar 

  • Miernyk, J.A., Dennis, D.T.: Mitochondrial, plastid, and cytosolic isozyme complexes of hexokinase from developing endosperm of Ricinus communis.-Arch. Biochem. Biophys. 226: 458–468, 1983.

    Article  PubMed  CAS  Google Scholar 

  • Renz, A., Merlo, L., Stitt, M.: Partial purification from potato tubers of three fructokinases and three hexokinases which show differing organ and developmental specificity.-Planta 190: 156–165, 1993.

    CAS  Google Scholar 

  • Roland, F., Winderickx, J., Thevelein, J.M.: Glucose-sensing mechanisms in eukaryotic cells.-Trends Biochem. Sci. 26: 310–317, 2001.

    Article  Google Scholar 

  • Schnarrenberger, C.: Characterization and compartmentation, in green leaves, of hexokinases with different specifities for glucose, fructose, and mannose and for nucleoside triphosphates.-Planta 181: 249–255, 1990.

    Article  CAS  Google Scholar 

  • Sheen, J., Zhou, L., Jang, J.-C.: Sugars as signaling molecules.-Curr. Opinion Plant Biol. 2: 410–418, 1999.

    Article  CAS  Google Scholar 

  • Šindelár, L., Makovcová, O.: Beziehungen zwischen der Phosphatasen-Aktivität und dem Gehalt der freien Zucker bei durch die Strichelkrankheit der Kartoffeln infiziertem N. tabacum cv. “Samsun”.-Biol. Plant. 16: 376–381, 1974.

    Article  Google Scholar 

  • Šindelár, L., Šindelárová, M., Burketová, L.: Changes in glucose, fructose and saccharose metabolism in tobacco plants infected with potato virus Y.-Biol. Plant. 42: 431–439, 1999.

    Article  Google Scholar 

  • Šindelárová, M., Šindelár, L.: Hexokinases of tobacco leaves: Subcellular localization and characterization.-Biol. Plant. 30: 275–284, 1988.

    Google Scholar 

  • Šindelárová, M., Šindelár, L.: Hexokinases of tobacco leaves: Influence of plant age on particulate and soluble isozyme composition.-Biol. Plant. 40: 469–474, 1997/1998.

    Article  Google Scholar 

  • Šindelárová, M., Šindelár, L., Burketová, L.: Dynamic changes in the activities of glucose-6–phosphate dehydrogenase, ribulose bisphosphate carboxylase, and ribonuclease in tobacco leaves, leaf discs, and mesophyll protoplasts in relation to TMV multiplication.-Physiol. mol. Plant Pathol. 51: 99–109, 1997.

    Article  Google Scholar 

  • Smeekens, S.: Sugar-induced signal transduction in plants.-Annu. Rev. Plant Physiol. Plant mol. Biol. 51: 49–81, 2000.

    Article  PubMed  CAS  Google Scholar 

  • Smith, S.B., Taylor, M.A., Burch, L.R., Davies, H.V.: Primary structure and characterization of a cDNA clone of fructokinase from potato (Solanum tuberosum L. cv. Record).-Plant Physiol. 102: 95–106, 1993.

    Article  Google Scholar 

  • Stitt, M., Bulpin, P.V., apRees, T.: Pathway of starch breakdown in photosynthetic tissues of Pisum sativum.-Biochim. biophys. Acta 544: 200–214, 1978.

    PubMed  CAS  Google Scholar 

  • Taylor, C.B.: Sweet sensations.-Plant Cell 9: 1–4, 1997.

    Article  CAS  Google Scholar 

  • Técsi, L.I., Maule, A.J., Smith, A.M., Leegood, R.C.: Metabolic alterations in cotyledons of Cucurbita pepo infected by cucumber mosaic virus.-J. exp. Bot. 45: 1541–1551, 1994.

    Google Scholar 

  • Turner, J.F., Turner, D.H.: The regulation of glycolysis and the pentose phosphate pathway.-In: Davies, D.D. (ed.): The Biochemistry of Plants. Vol. 2. Pp. 279–316. Academic Press, New York-London-Toronto-Sydney-San Francisco 1980.

    Google Scholar 

  • Umemura, T., Perata, P., Futsuhara, Y., Yamaguchi, J.: Sugar sensing and a-amylase gene repression in rice embryos.-Planta 204: 420–428, 1998.

    Article  PubMed  CAS  Google Scholar 

  • Wiese, A., Groner, F., Sonnewald, U., Deppner, H., Lerchi, J., Hebbeker, U., Flügge, U.-I., Weber, A.: Spinach hexokinase I is located in the outer envelope membrane of plastids.-Fed. eur. biochem. Soc. Lett. 461: 13–18, 1999.

    CAS  Google Scholar 

  • Wolffgang, H., Keck, A.: Untersuchungen über den Stoffwechsel viruskranker Pflanzen. I. Die Phosphatase-Aktivität in Nicotiana tabacum L. var. Samsun nach Infektion mit TMV.-Phytopathol. Z. 34: 57–65, 1959.

    Google Scholar 

  • Xiao, W., Sheen, J., Jang, J.-C.: The role of hexokinase in plant sugar signal transduction and growth and development.-Plant mol. Biol. 44: 451–461, 2000.

    Article  PubMed  CAS  Google Scholar 

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Šindelář, L., Šindelářová, M. Hexokinases of Tobacco Leaves: Changes in the Cytosolic and Non-Cytosolic Isozyme Complexes Induced by Tobacco Mosaic Virus Infection. Biologia Plantarum 47, 413–419 (2003). https://doi.org/10.1023/B:BIOP.0000023885.99970.30

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  • DOI: https://doi.org/10.1023/B:BIOP.0000023885.99970.30

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