Abstract
A full-length cDNA clone encoding microbody NAD+-dependent malate dehydrogenase (MDH) of cucumber has been isolated. The deduced amino acid sequence is 97% identical to glyoxysomal MDH (gMDH) of watermelon, including the amino terminal putative transit peptide. The cucumber genome contains only a single copy of this gene. Expression of this mdh gene increases dramatically in cotyledons during the few days immediately following seed imbibition, in parallel with genes encoding isocitrate lyase (ICL) and malate synthase (MS), two glyoxylate cycle enzymes. The level of MDH, ICL and MS mRNAs then declines, but then MDH mRNA increases again together with that of peroxisomal NAD+-dependent hydroxypyruvate reductase (HPR). The mdh gene is also expressed during cotyledon senescence, together with hpr, icl and ms genes. These results indicate that a single gene encodes MDH which functions in both glyoxysomes and peroxisomes. In contrast to icl and ms genes, expression of the mdh gene is not activated by incubating detached green cotyledons in the dark, nor is it affected by exogenous sucrose in the incubation medium. The function of this microbody MDH and the regulation of its synthesis are discussed.
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References
Alani AA, Luster DG, Donaldson RP: Development of endoplasmic reticulum and glyoxysomal membrane redox activities during castor bean germination. Plant Physiol 94: 1842–1848 (1990).
ap Rees T: Carbon metabolism in mitochondria. In: Dennis DT, Turpin DH (eds) Plant Physiology, Biochemistry and Molecular Biology, pp. 106–123. Longman Scientific and Technical, Harlow (1990).
De Bellis L, Picciarelli P, Pistelli L, Alpi A: Localisation of glyoxylate-cycle marker enzymes in peroxisomes of senescent leaves and green cotyledons. Planta 180: 435–439 (1990).
De Bellis L, Nishimura M: Development of enzymes of the glyoxylate cycle during senescence of pumpkin cotyledons. Plant Cell Physiol 32: 555–561 (1991).
Douce R, Neuburger M: Metabolite exchange between the mitochondrion and the cytosol. In: Dennis DT, Turpin DH (eds) Plant Physiology, Biochemistry and Molecular Biology, pp. 173–190. Longman Scientific and Technical, Harlow (1990).
Draper J, Scott R, Armitage P, Walden R (eds): Plant Genetic Transformation and Gene Expression: A Laboratory Manual. Blackwell Scientific Publications, Oxford (1988).
Feinberg AP, Vogelstein B: A technique for radiolabelling DNA fragments to high specific activity. Analyt Biochem (Addendum) 137: 266–267 (1984).
Gietl C: Glyoxysomal malate dehydrogenase from watermelon is synthesised with an amino-terminal transit peptide. Proc Natl Acad Sci USA 87: 5773–5777 (1990).
Gietl C, Lehnerer M, Olsen O: Mitochondrial malate dehydrogenase from watermelon: sequence of cDNA clones and primary structure of the higher-plant precursor protein. Plant Mol Biol 14: 1019–1030 (1990).
Gietl C: Partitioning of malate dehydrogenase isoenzymes into glyoxysomes, mitochondria and chloroplasts. Plant Physiol 100: 557–559 (1992).
Graham IA, Smith LM, Brown JWS, Leaver CJ, Smith SM: The malate synthase gene of cucumber. Plant Mol Biol 13: 673–684 (1989).
Graham IA, Smith LM, Leaver CJ, Smith SM: Developmental regulation of expression of the malate synthase gene in transgenic plants. Plant Mol Biol 15: 539–549 (1990).
Graham IA, Leaver CJ, Smith SM: Induction of malate synthase gene expression in senescent and detached organs of cucumber. Plant Cell 4: 349–357 (1992).
Greenler JM, Sloan JS, Schwarzt BW, Becker WM: Isolation, characterisation, and sequence analysis of a full-length cDNA clone encoding NADH-dependent hydroxypyruvate reductase from cucumber. Plant Mol Biol 13: 139–150 (1989).
Halliwell B: Chloroplast Metabolism. clarendon Press, Oxford (1984).
Kim D-J, Smith SM: Molecular cloning of cucumber phosphoenolpyruvate carboxykinase and developmental regulation of gene expression. Plant Mol Biol 26: 423–434 (1994).
Kudielka RA, Theimer RR: Repression of glyoxysomal enzyme activities in anise (Pimpinella anisum L.) suspension cultures. Plant Sci Lett 31: 245–252 (1983).
Lado P, Schwendimann M, Marre E: Repression of isocitrate lyase synthesis in seeds germinated in the presence of glucose. Biochim Biophys Acta 157: 140–148 (1968).
Liu KDF, Huang AHC: Developmental studies of NAD-malate dehydrogenase isozymes in the cotyledon of cucumber seedlings grown in darkness and in light. Planta 131: 279–284 (1976).
Longo CP, Longo GP: The development of glyoxysomes in peanut cotyledons and maize scutella. Plant Physiol 45: 249–254 (1970).
McLaughlin JC: The regulation and synthesis of malate synthase and isocitrate lyase in senescent and detached cotyledons of cucumber (Cucumis sativus). Ph.D. thesis, University of Edinburgh (1994).
McLaughlin JC, Smith SM: Metabolic regulation of glyoxylate cycle enzyme synthesis in detached cucumber cotyledons and protoplasts. Planta, in press.
Mettler IJ, Beevers H: Oxidation of NADH in glyoxysomes by a malate-aspartate shuttle. Plant Physiol 66: 555–560 (1980).
Ogren WL: Photorespiration: pathways, regulation and modification. Annu Rev Plant Physiol 35: 415–442 (1984).
Pistelli L, De Bellis L, Alpi A: Peroxisomal enzyme activities in attached senescing leaves. Planta 184: 151–153 (1991).
Preisig-Müller R, Kindl H: Thiolase mRNA translated in vitro yields a peptide with a putative N-terminal presequence. Plant Mol Biol 22: 59–66 (1993)
Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Press, Cold Spring Harbor, NY (1989).
Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).
Sloan JS, Schwarz BW, Becker WM: Promoter analysis of a light-regulated gene encoding hydroxypyruvate reductase, an enzyme of the photorespiratory glycolate pathway. Plant J 3: 867–874 (1993).
Vincentini F, Matile P: Gerontosomes, a multifunctional type of peroxisome in senescent leaves. J Plant Physiol 142: 50–56 (1993).
Wanner LF, Keller F, Matile P: Metabolism of radiolabelled galactolipids in senescent barley leaves. Plant Sci 78: 199–206 (1991).
Weir EM, Reizman H, Grienbenberger JM, Becker WM, Leaver CJ: Regulation of glyoxysomal enzymes during germination of cucumber: temporal changes in translatable mRNAs for isocitrate lyase and malate synthase. Eur J Biochem 112: 469–477 (1980).
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Kim, DJ., Smith, S.M. Expression of a single gene encoding microbody NAD-malate dehydrogenase during glyoxysome and peroxisome development in cucumber. Plant Mol Biol 26, 1833–1841 (1994). https://doi.org/10.1007/BF00019496
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DOI: https://doi.org/10.1007/BF00019496