Summary
Cytofluorometric methods were used to study the syntheses of DNA and of some proteins during the second intermitotic period of isolated nuclei and nucleoli of Physarum polycephalum. Interferometric methods were used for the determination of their dry mass.
The volume and dry mass of nuclei and nucleoli showed similar behaviour, the nuclei doubling their volume and dry mass, and the nucleoli tripling them.
The DNA synthesis began immediately after mitosis; the content nearly doubled after the first 3 hours in the case of the nuclei, after the first 5 hours in the case of the nucleoli. In both cases a slow increase (10%) was registered until the next mitosis.
Of all proteins investigated only protein-bound lysine of nuclei and nucleoli did not double in initial content, this is ascribed to the isolation method used. This fact was also reflected by the total protein analysis, as nuclei and nucleoli were very rich in lysine. Protein-bound arginine of the nuclei and nucleoli showed an evolution similar to DNA. The histones showed a synthesis independent of that of DNA. The comparison of total protein content and dry mass suggested that the quicker increase of the latter should be attributed to RNA.
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References
Braun, R.: Personal communication.
Braun, R., Behrens, K.: A ribonuclease from Physarum, biochemical properties and synthesis in the mitotic cycle. Biochim. biophys. Acta (Amst.) 195, 87–98 (1969).
Braun, R., Evans, T. E.: Replication of nuclear satellite and mitochondrial DNA in the mitotic cycle of Physarum polycephalum. Biochim. biophys. Acta (Amst.) 182, 511–522 (1969).
Braun, K., Mittermayer, C., Rusch, H. P.: Sequential temporal replication of DNA in Physarum polycephalum. Proc. nat. Acad. Sci. (Wash.) 53, 924–931 (1965).
Daniel, J. W., Baldwin, H. H.: Methods of culture for plasmodial myxomycetes. In: Methods in cell physiology, vol. 1, ed. Prescott, D. M., p. 9–41. New York: Acad. Press Inc. 1964.
Guttes, E., Guttes, S.: Mitotic synchrony in the plasmodia of Physarum polycephalum and mitotic synchronisation by coalescence of microplasmodia, In: Methods in cell physiology, vol. 1, ed. Prescott, D. M., p. 43–54. New York: Acad. Press Inc. 1964.
Guttes, E., Guttes, S.: Replication of nucleolus-associated DNA during “G2-phase” in Physarum polycephalum. J. Cell Biol. 43, 229–236 (1969).
Killander, D.: Intercellular variations in generation time and amounts of DNA, RNA and mass in a mouse leukemia population in vitro. Exp. Cell Res. 40, 21–31 (1969).
Killander, D., Zetterberg, A.: Quantitative cytochemical studies on interphase growth. Exp. Cell Res. 38, 272–284 (1965).
Leemann, U.: Personal communication.
Leemann, U., Ruch, F.: Cytofluorometric determination of basic and total protein with sulfaflavine. In preparation.
Leemann, U., Weiss, S., Schmutz, E.: A centrifugation technique for cytological preparations J. Histochem. Cytochem. 19, 758–760 (1971).
Mittermayer, C., Braun, R., Rusch, H. P.: RNA synthesis in the mitotic cycle of Physarum polycephalum. Biochem. Biophys. Acta 91, 399–405 (1964).
Mittermayer, C., Braun, R., Rusch, H. P.: The effect of actinomycin D on the timing of mitosis in Physarum polycephalum. Exp. Cell Res. 38, 33–41 (1965).
Mohberg, J. E., Rusch, H. P.: Isolation of nuclei and histones from the plasmodia of Physarum polycephalum. J. Cell Biol. 23, 61A (1964).
Mohberg, J. E., Rusch, H. P.: Nuclear histones in Physarum polycephalum during growth and differentiation. Arch. Biochem. Biophys. 138, 418–432 (1970).
Rosselet, A.: Mikrofluorometrische Argininbestimmung. Z. Wiss. Mikr. 68, 22–41 (1967).
Rosselet, A., Ruch, F.: Cytofluorometric determination of lysine with dansylchloride. J. Histochem. Cytochem. 16, 459–466 (1968).
Ruch, F.: Determination of DNA content by microfluorometry. In: Introduction to quantitative cytochemistry, ed. Wied, G. L., p. 281–294. New York: Acad. Press Inc. 1966.
Zetterberg, A., Killander, D.: Quantitative Cytofluorometric and autoradiographic studies on the rate of protein synthesis during interphase in mouse fibroblasts in vitro. Exp. Cell Res. 40, 1–10 (1965).
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Bovey, F., Ruch, F. Cytofluorometric determination of DNA and protein in the nuclei and nucleoli of Physarum polycephalum during the intermitotic period. Histochemie 32, 153–162 (1972). https://doi.org/10.1007/BF00303730
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DOI: https://doi.org/10.1007/BF00303730