Abstract
The polysomal pattern of the dinoflagellate Gonyaulax polyedra, cultured under constant conditions, demonstrates a circadian rhythm. The relative amount of polysomes increases during the phase corresponding to the previous night period (=subjective night phase) when the rate of protein synthesis reaches its maximum (Cornelius et al., 1985, Planta 160, 365–370). Cell-free extracts were isolated at different circadian phase. The rate of protein synthesis in the extracts changed rhythmically in the same manner as the rate of protein synthesis in vivo. Substances in the postribosomal supernatants influenced the protein-synthesis rate of the cell-free system, depending on the phase when they were isolated: “night factors” stimulated protein synthesis in “day extracts” whereas “day factors” inhibited protein synthesis in “night extracts”. These effects were abolished by heating the postribosomal supernatant. In-vitro phosphorylation in parallel probes showed changes in the pattern of phosphorylated proteins. Phosphorylation of one of the proteins (95 kDa) was decreased after addition of “night factor(s)” and increased after addition of “day factor(s)”. Cyclic-AMP enhanced the rates of protein synthesis and phosphorylation in the day extracts.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- cAMP:
-
cyclic-AMP
- CT:
-
circadian time
- D (N):
-
subjective day (night)phase
References
Alton, T.H., Lodish, H.F. (1977) Translational control of protein synthesis during the early stages of differentiation of the slime mold Dictyostelium discoideum. Cell 12, 301–310
Cornelius, G., Schröder-Lorenz, A., Rensing, L. (1985) Circadian clock control of protein synthesis and degradation in Gonyaulax polyedra. Planta 160, 365–370
Cummings, F.W. (1975) A biochemical model of the circadian clock. J. Theor. Biol. 55, 455–470
Donner, B., Helmboldt-Caesar, U., Rensing, L. (1985) Circadian rhythm of total protein synthesis in the cytoplasm and chloroplasts of Gonyaulax polyedra. Chronobiol. Int. 2, 1–9
Duncan, R., McConkey, E.H. (1982) Rapid alterations in initiation rate and recruitment of inactive RNA are temporally correlated with S6 phosphorylation. Eur. J. Biochem. 123, 539–544
Dunlap, J.C., Taylor, W., Hastings, J.W. (1980) The effects of protein synthesis inhibitors on the Gonyaulax clock. I. Phase shifting effects of cycloheximide. J. Comp. Physiol. 238, 1–8
Ernst, V., Levin, D.H., Foulkes, J.G., London, I.M. (1982) Effects of skeletal muscle protein phosphatase inhibitor-2 on protein synthesis and protein phosphorylation in rabbit reticulocyte lysates. Proc. Natl. Acad. Sci. USA 79, 7092–7096
Ernst, V., Levin, D.H., Ranu, R.S., London, I.M. (1976) Control of protein synthesis in reticulocyte lysates: Effects of 3′: 5′-cyclic AMP, ATP and GTP on inhibitions induced by heme-deficiency, double-stranded RNA, and a reticulocyte translational inhibitor. Biochemistry 73, 1112–1116
Eskin, A., Takahashi, J.S. (1983) Adenylate cyclase activation shifts the phase of a circadian pacemaker. Science 220, 82–84
Eskin, A., Yeung, S.J., Klass, M.R. (1984) Requirement for protein synthesis in the regulation of a circadian rhythm by serotonin. Proc. Natl. Acad. Sci. USA 81, 7637–7641
Farrell, P.J., Balkow, K., Hunt, T., Jackson, R.J., Trachsel, H. (1977) Phosphorylation of initiation factor elF2 and the control of reticulocyte protein synthesis. Cell 11, 187–200
Guillard, R.R.L., Ryther, J.H. (1962) Studies of marine planktonic diatoms. I. Cyclotella nana Hustedt and Detonula confervacea (Cleve) Grand. Can. J. Microbiol. 8, 229–239
Laemmli, U.K. (1970) Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227, 680–685
Mans, R.J., Novelli, G.P. (1961) Measurement of the incorporation of radioactive amino acids into protein by a filter-disk method. Arch. Biochem. Biophys. 94, 48–53
Neuhoff, V.K., Philipp, H., Zimmer, G., Mesecke, S. (1979) A simple, versatile, sensitive and volume independent method for quantitation protein determination, which is independent of other external influences. Hoppe-Seyler's Z. Physiol. Chem. 360, 1657–1670
Ranu, R.S. (1980) Isolation of a translational inhibitor from wheat germ with protein kinase activity that phosphorylates initiation factor eIF2. Biochem. Biophys. Res. Commun. 97, 1124–1132
Ranu, R.S., London, I.M. (1979) Regulation of protein synthesis in rabbit reticulocyte lysates: Additional initiation factor required for formation of ternary complex (eIF2·GTP·Mett RNA) and demonstration of inhibitory effect of hemeregulated protein kinase. Proc. Natl. Acad. Sci. USA 76, 1079–1083
Rensing, L., Schill, W. (1985) Perturbation by single and double pulses as analytical tool for analyzing oscillatory mechanisms. In: Temporal order, pp. 224–225 Rensing, L., Jaeger, N.I., eds. Springer, Berlin Heidelberg New York
Schroeder-Lorenz, A., Rensing, L. (1986) Circadian clock mechanism and synthesis rates of individual protein species in Gonyaulax polyedra. Comp. Biochem. Physiol 85B, 315–323
Taylor, W.R., Dunlap, J.C., Hastings, J.W. (1982) Inhibitors of protein synthesis on 80 S ribosomes phase shift the Gonyaulax clock. J. Exp. Biol. 97, 121–136
Volknandt, W., Hardeland, R. (1984) Circadian rhythmicity of protein synthesis in the dinoflagellate, Gonyaulax polyedra: A biochemical and radioautographic investigation. Comp. Biochem. Physiol. 77B, 493–500
Weber, K., Osborn, M. (1969) The reliability of molecular weight determinations by dodecyl sulfate polyacrylamide gel electrophoresis. J. Biol. Chem. 244, 4406–4412
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Schröder-Lorenz, A., Rensing, L. Circadian changes in protein-synthesis rate and protein phosphorylation in cell-free extracts of Gonyaulax polyedra . Planta 170, 7–13 (1987). https://doi.org/10.1007/BF00392374
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00392374