Summary
Experiments were designed to determine if the synthesis, processing and transport of rRNA are affected by changes in nuclear permeability. RNA was labeled by microinjecting [3H]GTP into the cytoplasm of defolliculated oocytes. After injection, the nuclear envelopes were disrupted by puncturing the cells with glass needles. It has been shown that this procedure significantly alters the physical properties of the nuclear envelope. At appropriate intervals after puncturing, the oocytes were manually enucleated and RNA was extracted from both nuclear and cytoplasmic fractions. The extracts were analyzed on 2.5% polyacrylamide gels. The results indicate that over a period of 41/2 h neither the production nor the nucleocytoplasmic distribution of rRNA are affected by altering the permeability characteristics of the nuclear envelope.
References
Anderson, D.M., Smith, L.D.: Synthesis of heterogeneous nuclear RNA in full-grown oocytes of Xenopus laevis (Daudin). Cell 11, 663–671 (1977)
Clawson, G.A., Smuckler, E.A.: Activation energy for RNA transport from isolated rat liver nuclei. Proc. Natl. Acad. Sci. USA 75, 5400–5404 (1978)
Clawson, G.A., Koplitz, M., Castler-Schechter, B., Smuckler, E.A.: Energy utilization and RNA transport: Their interdependence. Biochemistry 17, 3747–3752 (1978)
Dumont, J.N.: Oogenesis in Xenopus laevis (Daudin). 1. Stages of oocyte development in laboratory maintained animals. J. Morphol. 136, 153–178 (1972)
Feldherr, C.M., Pomerantz, J.: Mechanism for the selection of nuclear polypeptides in Xenopus oocytes. J. Cell Biol. 78, 168–175 (1978)
Feldherr, C.M., Richmond, P.A.: Manual enucleation of Xenopus oocytes. In: Methods in Cell Biology (G. Stein, J. Stein, L. J. Kleinsmith, eds.), Vol. 17, pp. 75–79. New York and London: Academic Press 1978
Franke, W.W., Scheer, U.: Pathways of nucleocytoplasmic translocation of ribonucleoproteins. In: Symposia of the Society for Experimental Biology, (M.A. Sleigh, D.H. Jennings, eds.), No. 28, pp. 249–282. Cambridge: University Press 1974
Herlan, G., Eckert, W.A., Kaffenberger, W., Wunderlich, F.: Isolation and characterization of an RNAcontaining nuclear matrix from Tetrahymena macronuclei. Biochemistry 18, 1782–1788 (1979)
Kanno, Y., Loewenstein, W.R.: A study of the nucleus and cell membranes of oocytes with an intracellular electrode. Exp. Cell Res. 31, 149–166 (1963)
Loening, U.E.: The fractionation of high-molecular-weight ribonucleic acid by polyacrylamide-gel electrophoresis. Biochem. J. 102, 251–257 (1967)
Stuart, S.E., Clawson, G.A., Rottman, F.M., Patterson, R.J.: RNA transport in isolated myeloma nuclei: Transport from membrane-denuded nuclei. J. Cell Biol. 72, 57–66 (1977)
Wunderlich, F., Berezney, R., Kleinig, H.: The nuclear envelope: An interdisciplinary analysis of its structure, composition, and functions. In: Biological Membranes, (D. Chapman, D.F.H. Wallach, eds.), Vol. 3, pp. 241–333. New York and London: Academic Press 1976
Author information
Authors and Affiliations
Additional information
Supported by grant GM 21531 from the NIH
The author would like to thank Mr. Jay Pomerantz for his excellent technical assistance, and also Drs. R.J. Cohen and P. McGuire for their interest and criticism
Rights and permissions
About this article
Cite this article
Feldherr, C.M. Ribosomal RNA synthesis and transport following disruption of the nuclear envelope. Cell Tissue Res. 205, 157–162 (1980). https://doi.org/10.1007/BF00234451
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00234451