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. 1978 Nov;75(11):5400–5404. doi: 10.1073/pnas.75.11.5400

Activation energy for RNA transport from isolated rat liver nuclei.

G A Clawson, E A Smuckler
PMCID: PMC392971  PMID: 281689

Abstract

The temperature dependence of ATP-enhanced RNA delivery from rat liver nuclei to a surrogate cytoplasm was investigated. Examination of linear-rate data on Arrhenius graphs of 1/T vs. log (% RNA delivered per min) revealed an activation energy of 12.5--13 kcal/mol. When data derived from longer incubation periods was displayed on Arrhenius graphs, we observed a discontinuous graph--two distinct linear segments with slopes of differing sign which intersected near 20 degrees C. It was demonstrated that this discontinuity was not due to lipid phase transition in the nuclear membranes and that its position depended upon treatment of the nuclei and upon additives to the incubation mixtures. The decline in transport apparent in the upper-temperature domain on 20-min Arrhenius graphs was shown to be based on the diffusion of transported macromolecular RNA back into the nucleus--a process greatly amplified by the rapidity of transport in this domain. The large net inward diffusion, in concert with significantly differing activation energies for RNA transport and passive diffusion, suggests that the process of nucleocytoplasmic RNA transport is not diffusion driven. Our data have established that an integral parameter of RNA transport (namely, the activation energy) remains unchanged in various in vitro manipulations.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Agutter P. S., McArdle H. J., McCaldin B. Evidence for involvement of nuclear envelope nucleoside triphosphatase in nucleocytoplasmic translocation of ribonucleoprotein. Nature. 1976 Sep 9;263(5573):165–167. doi: 10.1038/263165a0. [DOI] [PubMed] [Google Scholar]
  2. Bertoli E., Parenti-Castelli G., Landi L., Sechi A. M., Lenaz G. Activation energies of mitochondrial adenosine triphosphatase under different conditions. J Bioenerg. 1973;4(6):591–598. doi: 10.1007/BF01516210. [DOI] [PubMed] [Google Scholar]
  3. Blobel G., Potter V. R. Nuclei from rat liver: isolation method that combines purity with high yield. Science. 1966 Dec 30;154(3757):1662–1665. doi: 10.1126/science.154.3757.1662. [DOI] [PubMed] [Google Scholar]
  4. Chatterjee N. K., Weissbach H. Release of RNA from HeLa cell nuclei. Arch Biochem Biophys. 1973 Jul;157(1):160–167. doi: 10.1016/0003-9861(73)90401-3. [DOI] [PubMed] [Google Scholar]
  5. Ishikawa K., Kuroda C., Ogata K. Messenger ribonucleoprotein complexes released from rat liver nuclei by ATP. II. Chemical and metabolic properties of the protein moiety of messenger ribonucleoprotein complexes. Biochim Biophys Acta. 1970 Aug 8;213(2):505–512. doi: 10.1016/0005-2787(70)90057-2. [DOI] [PubMed] [Google Scholar]
  6. Lenk R., Ransom L., Kaufmann Y., Penman S. A cytoskeletal structure with associated polyribosomes obtained from HeLa cells. Cell. 1977 Jan;10(1):67–78. doi: 10.1016/0092-8674(77)90141-6. [DOI] [PubMed] [Google Scholar]
  7. Nägel W. C., Wunderlich F. Effect of temperature on nuclear membranes and nucleo-cytoplasmic RNA-transport in Tetrahymena grown at different temperatures. J Membr Biol. 1977 Apr 7;32(1-2):151–164. doi: 10.1007/BF01905214. [DOI] [PubMed] [Google Scholar]
  8. Racevskis J., Webb T. E. Processing and release of ribosomal RNA from isolated nuclei: analysis of the ATP-dependence and cytosol-dependence. Eur J Biochem. 1974 Nov 1;49(1):93–100. doi: 10.1111/j.1432-1033.1974.tb03814.x. [DOI] [PubMed] [Google Scholar]
  9. Raison J. K. The influence of temperature-induced phase changes on the kinetics of respiratory and other membrane-associated enzyme systems. J Bioenerg. 1973 Jan;4(1):285–309. doi: 10.1007/BF01516063. [DOI] [PubMed] [Google Scholar]
  10. Raskas H. J., Rho Y. C. ATP requirement for release of adenovirus mRNA from isolated nuclei. Nat New Biol. 1973 Sep 12;245(141):47–49. doi: 10.1038/newbio245047a0. [DOI] [PubMed] [Google Scholar]
  11. Roth H. E., Bolla R., Cox G. S., Redfield B., Weissbach H., Brot N. Uptake of ribosomal proteins by isolate HeLa nuclei. Biochem Biophys Res Commun. 1976 Apr 5;69(3):608–612. doi: 10.1016/0006-291x(76)90919-0. [DOI] [PubMed] [Google Scholar]
  12. SCHNEIDER J. H. Factors affecting the release of nuclear ribonucleic acid from the nucleus in vitro. J Biol Chem. 1959 Oct;234:2728–2732. [PubMed] [Google Scholar]
  13. Sato T., Ishikawa K., Ogata K. Characterization of ribonucleoprotein particles released from isolated nuclei of regenerating rat liver in two different in vitro systems. Biochim Biophys Acta. 1977 Feb 16;474(4):536–548. doi: 10.1016/0005-2787(77)90074-0. [DOI] [PubMed] [Google Scholar]
  14. Sauermann G. Nuclear columns: release of ribonucleoproteins from rat liver nuclei. Biochem Biophys Res Commun. 1974 Jan;56(1):155–180. doi: 10.1016/s0006-291x(74)80328-1. [DOI] [PubMed] [Google Scholar]
  15. Shearer R. W., Smuckler E. A. Altered regulation of the transport of RNA from nucleus to cytoplasm in rat hepatoma cells. Cancer Res. 1972 Feb;32(2):339–342. [PubMed] [Google Scholar]
  16. Smuckler E. A., Koplitz M. Thioacetamide-induced alterations in nuclear RNA transport. Cancer Res. 1974 Apr;34(4):827–838. [PubMed] [Google Scholar]
  17. Smuckler E. A., Koplitz R. M. Polyadenylic acid content and electrophoretic behavior of in vitro released RNA's in chemical carcinogenesis. Cancer Res. 1976 Mar;36(3):881–888. [PubMed] [Google Scholar]
  18. 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. 1977 Jan;72(1):57–66. doi: 10.1083/jcb.72.1.57. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Stuart S. E., Rottman F. M., Patterson R. J. Nuclear restriction of nucleic acids in the presence of ATP. Biochem Biophys Res Commun. 1975 Jan 20;62(2):439–447. doi: 10.1016/s0006-291x(75)80158-6. [DOI] [PubMed] [Google Scholar]
  20. Wise G. E., Goldstein L. Electron microscope localization of nuclear RNA's that shuttle between cytoplasm and nucleus and nuclear RNA's that do not. J Cell Biol. 1973 Jan;56(1):129–138. doi: 10.1083/jcb.56.1.129. [DOI] [PMC free article] [PubMed] [Google Scholar]

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