Abstract
The production of poly (A)+ RNA was determined over the cell-cycle in Euglena cultures division-synchronized by 14 h light periods alternating with 10 h dark periods. The amount of poly (A)+ RNA per culture aliquot was constant from the beginning until near the end of the light phase of the cell-cycle when it increased in a sharp burst after the time of DNA replication. Poly (A)+ RNA from all stages of the cell-cycle promoted protein synthesis with equal efficiency in a mRNA-dependent reticulocyte lysate translation system. Malate synthase mRNA was detected using malate synthase antibody to precipitate nascent malate synthase protein in the translation system, binding the IgG-antigen complex to Staphylococcus aureus, concentrating, eluting the IgG-antigen complex and analysing the products by sodium dodecyl sulfatepolyacrylamide gel electrophoresis. Malate synthase mRNA was absent from phototrophic cells and was only produced in darkened cultures following the addition of acetate.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- TMV:
-
tobacco mosaic virus
- SMISH buffer:
-
10 mM Tris-HCl pH 7.2, containing 25 mM KCl, 10 mM NaCl, 1 mM MgCl2, and 0.2 mM dithiothreitol
- SDS:
-
Sodium dodecyl sulfate
- TCA:
-
trichloroacetic acid
References
Codd, G. A.: Carbon dioxide metabolism in synchronously dividing cultures of Euglena gracilis. Ph. D. thesis. University of Bradford, 1970
Codd, G. A., Merrett, M. J.: Photosynthetic products of division synchronized cultures of Euglena. Plant Physiol. 47, 635–639 (1971)
Collins, N., Merrett, M. J.: Microbody-marker enzymes during transition from phototrophic to organotrophic growth in Euglena. Plant Physiol. 55, 1018–1022 (1075)
Cramer, M., Myers, J.: Growth and photosynthetic characteristics of Euglena gracilis. Arch. Mikrobiol. 17, 384–402 (1952)
Craig, R. K., Brown, P. A., Harrison, O. S., McIlrevy, D., Campbell, P.: Guinea-Pig Milk-Protein Synthesis. Isolation and characterization of messenger ribonucleic acids from lactating mammary gland and identification of caseins and pre-α-lactalbumin as translation products in heterologous cell-free systems. Biochem. J. 160, 57–74 (1976)
Davis, B., Merrett, M. J.: The effect of light on the synthesis of mitochondrial enzymes in division-synchronized Euglena cultures. Plant Physiol. 53, 575–580 (1974)
Fellig, J., Wiley, C. E.: Ribonuclease of Euglena gracilis. Science 132, 1835–1836 (1960)
Forde, B. G., John, P. C. L.: Stepwise accumulation of autoregulated enzyme activities during the cell cycle of the eucaryotic Chlorella. Exp. Cell. Res. 79, 127–135 (1973)
Gooding, G. V., Herbert, T. T.: A simple technique for purification of tobacco mosaic virus in large quantities. Phytopathology 57, 1285 (1967)
Hunt, T., Jackson, R. J.: The rabbit reticulocyte lysate as a system for studying m RNA. In: Modern trends in human leukaemia (R. Netts, R. C. Gallo, S. Spiegelman, F. Stohlman, eds.), pp. 300–307. München: J. F. Lehmanns 1974
Kessler, S. W.: Rapid isolation of antigens from cells with a Straphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J. Immunol. 115, 1617–1624 (1975)
Mans, R. J., Novelli, G. D.: Measurement of the incorporation of radioactive amino-acids into protein by a filter-paper disk method. Arch. Biochem. Biophys. 94, 48–53 (1961)
Parish, J. H., Kirby, K. S.: Reagents which reduce interactions between ribosomal RNA and rapidly labelled RNA from rat liver. Biochim. Biophys. Acta 129, 554–562 (1966)
Pelham, H. R. B., Jackson, R. J.: An efficient mRNA-dependent translation system from reticulocyte lysates. Eur. Biochem. 67, 247–256 (1976)
Perry, R. P., Le Torre, J., Kelley, D. E., Greenberg, J. R.: On the lability of poly (A) sequences during extraction of messenger RNA from polyribosomes. Biochim. Biophys. Acta 262, 220–226 (1972)
Rogg, H., Wehrli, W., Staehlin, M.: Isolation of mammalian transfer RNA. Biochim. Biophys. Acta 195, 13–15 (1969)
Sagher, D., Edelman, M., Jakob, K. M.: Poly (A)+ associated RNA in plants. Biochim. Biophys. Acta 349, 32–38 (1974)
Scragg, A. H., John, P. C. L., Thurston, C. F.: Post-transcription control of isocitrate lyase induction in the eukaryotic alga. Chlorella fusca. Nature 257, 498–501 (1975)
Thurston, C. F.: Control of isocitrate lyase synthesis in Chlorella fusca var. vacuolata. The basal activity of the enzyme and kinetics of induction. Biochem. J. 164, 147–151 (1977)
Verdier, G., Trabuchet, G., Heizmann, P., Nigon, P.: Effect l'éclairement sur les synthèses de RNA et de séquences polyadenyliques dans les cultures d'Euglena gracilis etiolées. Biochim. Biophys. Acta 312, 528–539 (1973)
Verdier, G.: Synthesis and translation site of light-induced mRNA's in etiolated Euglena gracilis. Biochim. Biophys. Acta 401, 91–98 (1975)
Woodcock, E., Merrett, M. J.: Purification and immunochemical characterization of malate synthase from Euglena gracilis. Biochem. J. 173, 95–101 (1978)
Woodward, J., Merrett, M. J.: Induction potential for glyoxylate cycle enzymes during the cell-cycle of Euglena gracilis. Eur. J. Biochem. 55, 555–559 (1975)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Woodcock, E., Merrett, M.J. Malate synthase messenger RNA in relation to enzyme derepression in Euglena gracilis . Arch. Microbiol. 124, 33–38 (1980). https://doi.org/10.1007/BF00407025
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00407025