Abstract
The development of peroxisomal enzymes in cotyledons of cucumber seedlings is strongly dependent on light. In light-grown seedlings, activities of two peroxisomal enzymes, hydroxypyruvate reductase (HPR) and serine: glyoxylate aminotransferase (SGAT), were barely detectable until three days postimbibition, after which time both activities increased rapidly and linearly for at least three days. In the dark, the activities of these enzymes increased slightly over the same time period, but only to about 5% to 10% of 7-day light-induced levels. When 51/2-day dark-grown seedlings were transferred into white light, activities of HPR and SGAT began to increase after approximately 8 h. HPR protein was shown by an immunoprecipitation assay to increase concurrently with enzymatic activity in both light- and dark-grown cotyledons. Immunoblotting results suggested that the amounts of SGAT-A and SGAT-B, the two subunits of SGAT, also developed along with SGAT activity. The relative levels of translatable mRNAs encoding HPR, SGAT-A, and SGAT-B were also light-dependent, and increased with a developmental pattern similar to enzyme activity and protein levels in light- and dark-grown cotyledons. In 51/2-day dark-grown cotyledons that were transferred to the light, translatable mRNAs for SGAT-A and SGAT-B began to increase within 1 h of illumination and continued of increase rapidly and linearly for the next 24 h in the light to a new steady-state level that was 45 times that of dark controls. Translatable HPR mRNA exhibited a biphasic pattern of accumulation, with a three-fold increase during the first 6 h of illumination, followed by an additional six-fold increase between 8 and 24 h. The accumulation of translationally active mRNA for both enzymes preceded the accumulation of the corresponding protein and enzyme activity by about 8 h. Our data suggest that the rise in enzyme activity depends on an increase in translatable mRNA for these enzymes and is regulated at a pretranslational level, most likely involving transcription of new mRNA.
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References
Apel K: Phytochrome-induced appearance of mRNA activity for the apoprotein of the light-harvesting chlorophyll a/b protein of barley (Hordeum vulgare). Eur J Biochem 97: 183–188 (1979).
Becker WM, Leaver CJ, Weir EM, Riezman H: Regulation of glyoxysomal enzymes during germination of cucumber. 1. Developmental changes in cotyledonary protein, RNA, and enzyme activities during germination. Plant Physiol 62: 542–549 (1978).
Becker WM, Riezman H, Weir EM, Titus DE, Leaver CJ: In vitro synthesis and compartmentalization of glyoxysomal enzymes in cucumber. Ann NY Acad Sci 386: 329–348 (1982).
Berger SL, Birkenmeier CS: Inhibition of intractable nucleases with ribonucleoside-vanadyl complexes: Isolation of messenger ribonucleic acid from resting lymphocytes. Biochemistry 18: 5143–5149 (1979).
Burnette WN: “Western Blotting”: electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radiolabeled protein A. Anal Biochem 112: 195–203 (1981).
Colbert JT, Hershey HP, Quail PH: Autoregulatory control of translatable phytochrome mRNA levels. Proc Natl Acad Sci USA 80: 2248–2252 (1983).
Feierabend J, Beevers H: Developmental studies on microbodies in wheat leaves. I. Conditions influencing enzyme development. Plant Physiol 49: 28–32 (1972).
Feierabend J: Developmental studies on microbodies in wheat leaves. III. On the photocontrol of microbody development. Planta 123: 63–77 (1975).
Gerhardt B: Studies on formation of glycolate oxidase in developing cotyledons of Helianthus annuus L. and Sinapis alba L. Z Pflanzenphysiol 74: 14–21 (1974).
Hoagland DR, Snyder WC: Nutrition of strawberry plant under controlled conditions. (a) Effects of deficiencies of boron and certain other elements. (b) Susceptibility to injury from sodium salts. Proc Amer Soc Hort Sci 30: 288–293 (1933).
Hondred D, Hunter JMcC, Keith R, Titus DE, Becker WM: Isolation of serine:glyoxylate aminotransferase from cucumber cotyledons. Plant Physiol 79: 95–102 (1985).
Hondred D, Titus DE, Wadle D-M, Becker WM: Regulation of peroxisomal enzymes in cotyledons of germinating cucumber seeds. Plant Physiol 75s: 143 (1984).
Hondred D, Titus DE, Wadle D-M, Becker WM: Light regulated appearance of peroxisomal enzymes in cucumbers. J Cell Biol 101: 79a (1985).
Hong UN, Schopfer P: Control by phytochrome of urate oxidase and allantoinase activities during peroxisome development in the cotyledons of mustard (Sinapis alba L.) seedlings. Planta 152: 325–335 (1981).
Huang AHC, Trelease RN, Moore TS: Plant Peroxisomes. Academic Press, New York (1983).
Kagawa T, Lord JM, Beevers H: Development of enzymes in the cotyledons of watermelon seedlings. Plant Physiol 51: 66–71 (1973).
Kagawa T, Beevers H: The development of microbodies (glyoxysomes and leaf peroxisomes) in cotyledons of germinating seedlings. Plant Physiol 55: 258–264 (1975).
Kessler SW: Rapid isolation of antigens from cells with staphylococcal protein A-antibody adsorbant: Parameters of the interaction of antibody-antigen complexes with protein A. J Immunol 115: 1617–1624 (1975).
Knecht DA, Dimond RL: Lysosomal enzymes possess a common antigenic determinant in the cellular slime mold, Dictyostelium discoideum. J Biol Chem 256: 3564–3575 (1981).
Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685 (1970).
Lamb JE, Riezman H, Leaver CJ, Becker WM: Regulation of glyoxysomal enzymes during germination of cucumber. 2. Isolation and immunological detection of isocitrate lyase and catalase. Plant Physiol 62: 754–760 (1978).
Laskey RA: The use of intensifying screens or organic scintillators for visualizing radioactive molcules resolved by gel electrophoresis. Meth Enzym 65: 363–371 (1980).
Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning. A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).
Mohr H: Pattern specification and realization in photomorphogenesis. Encyclopedia of Plant Physiol (NS) 16A: 336–357 (1983).
Murray MG, Peters DL, Thompson WF: Ancient repeated sequences in the pea and mung bean genomes and implications for genome evolution. J Mol Evol 17: 31–42 (1981).
Nelson T, Harpster MH, Mayfield SP, Taylor WC: Light-regulated gene expression during maize leaf development. J Cell Biol 98: 558–564 (1984).
Pelham HRB, Jackson RJ: An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem 67: 247–256 (1976).
Rehfeld DW, Tolbert NE: Aminotransferases in peroxisomes from spinach leaves. J Biol Chem 247: 4803–4811 (1972).
Schnarrenberger C, Oeser A, Tolbert NE: Development of microbodies in sunflower cotyledons and castor bean endosperm during germination. Plant Physiol 48: 566–574 (1971).
Schopfer P, Bajracharya D, Bergfeld R, Falk H: Phytochrome-mediated transformation of glyoxysomes into peroxisomes in the cotyledons of mustard (Sinapis alba) seedlings. Planta 133: 73–80 (1976).
Smith SM, Leaver CJ: Clyoxysomal malate synthase of cucumber: Molecular cloning of a cDNA and regulation of enzyme synthesis during germination. Plant Physiol 81: 762–767 (1986).
Titus DE, Hondred D, Becker WM: Purification and characterization of hydroxypyruvate reductase from cucumber cotyledons. Plant Physiol 72: 402–408 (1983).
Tobin EM: White light effects on the mRNA for the light-harvesting chloropyll a/b-protein in Lemna gibba L. G-3. Plant Physiol 67: 1078–1083 (1981).
Tobin EM, Silverthorne J: Light regulation of gene expression in higher plants. Ann Rev Plant Physiol 36: 569–593 (1985).
Tobin EM, Suttie JL: Light effect on the synthesis of ribulose-1,5-bisphosphate carboxylase in Lemna gibba L. G-3. Plant Physiol 65: 641–647 (1980).
Tokuhisa JG, Daniels SM, Quail PH: Phytochrome in green tissue: Spectral and immunochemical evidence for two distinct molecular species of phytochrome in light-grown Avena sativa L. Planta 164: 321–332 (1985).
Trelease RN, Becker WM, Gruber PJ, Newcomb EH: Microbodies (glyoxysomes and peroxisomes) in cucumber cotyledons. Correlative biochemical and ultrastructural study in light- and dark-grown seedlings. Plant Physiol 48: 461–475 (1971).
Vincent A, Akhayat O, Goldenberg S, Scherrer K: Differential repression of specific mRNA in erythroblast cytoplasm: A possible role for free mRNPA proteins. EMBO J 2: 1869–1876 (1983).
Weir EM, Riezman H, Grienenberger JM, Becker WM, Leaver CJ: Regulation of glyoxysomal enzymes during germination of cucumber. 4. Temporal changes in translatable mRNAs for isocitrate lyase and malate synthase. Eur J Biochem 112: 469–477 (1980).
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Hondred, D., Wadle, DM., Titus, D.E. et al. Light-stimulated accumulation of the peroxisomal enzymes hydroxypyruvate reductase and serine:glyoxylate aminotransferase and their translatable mRNAs in cotyledons of cucumber seedlings. Plant Mol Biol 9, 259–275 (1987). https://doi.org/10.1007/BF00166462
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DOI: https://doi.org/10.1007/BF00166462