Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1968 Apr 1;37(1):163–181. doi: 10.1083/jcb.37.1.163

ISOLATION OF TWO DISTINCT CLASSES OF POLYSOMES FROM A NUCLEAR FRACTION OF RAT LIVER

Paul D Sadowski 1, Janet Alcock Howden 1
PMCID: PMC2107385  PMID: 5645839

Abstract

Isolated rat liver nuclei were washed with Triton-X-100 in the presence of liver cell sap. This treatment liberated a fraction of polysomes which were isolated by differential centrifugation and were designated "outer membrane polysomes." The outer membrane polysomes synthesized protein in vivo. Shortly after injection of orotic acid-14C, the RNA of outer membrane polysomes had a higher specific activity than that of cytoplasmic polysomes. It was postulated that outer membrane polysomes may be an intermediate in the transfer of newly synthesized RNA from the nucleus to the cytoplasm. In other experiments, Triton-washed rat liver nuclei were lysed in the presence of deoxycholate and deoxyribonuclease. A ribonucleoprotein fraction was isolated from the lysate by differential centrifugation. This fraction contained "intranuclear ribosomes," which sedimented like partially degraded polysomes in sucrose gradients. This degradation could be partially prevented if intranuclear ribosomes were purified by sedimentation through heavy sucrose. The resulting pellets were termed "intranuclear polysomes" because they contained some undergraded polysomes. Intranuclear polysomes were highly radioactive after a brief pulse with orotic acid-14C, but did not appear to synthesize protein rapidly in vivo. Intranuclear polysomes may represent the initial stage of assembly of polyribosomes in the nucleus.

Full Text

The Full Text of this article is available as a PDF (1.6 MB).

Selected References

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

  1. BIRNSTIEL M. L., CHIPCHASE M. I., HYDE B. B. THE NUCLEOLUS, A SOURCE OF RIBOSOMES. Biochim Biophys Acta. 1963 Nov 22;76:454–462. [PubMed] [Google Scholar]
  2. BREMER H., KONRAD M. W. A COMPLEX OF ENZYMATICALLY SYNTHESIZED RNA AND TEMPLATE DNA. Proc Natl Acad Sci U S A. 1964 May;51:801–808. doi: 10.1073/pnas.51.5.801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BYRNE R., LEVIN J. G., BLADEN H. A., NIRENBERG M. W. THE IN VITRO FORMATION OF A DNA-RIBOSOME COMPLEX. Proc Natl Acad Sci U S A. 1964 Jul;52:140–148. doi: 10.1073/pnas.52.1.140. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bach M. K., Johnson H. G. Polysomes on a nuclear membrane fraction as intermediates in the transfer of ribonucleic acid from the nucleus to the cytoplasm. Nature. 1966 Feb 26;209(5026):893–895. doi: 10.1038/209893a0. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Blobel G., Potter V. R. Relation of ribonuclease and ribonuclease inhibitor to the isolation of polysomes from rat liver. Proc Natl Acad Sci U S A. 1966 May;55(5):1283–1288. doi: 10.1073/pnas.55.5.1283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. CHAUVEAU J., MOULE Y., ROUILLER C. Isolation of pure and unaltered liver nuclei morphology and biochemical composition. Exp Cell Res. 1956 Aug;11(2):317–321. doi: 10.1016/0014-4827(56)90107-0. [DOI] [PubMed] [Google Scholar]
  8. ELAEV N. R., RYKHLIK I. SRAVNITEL'NOE ISSLEDOVANIE TSITOPLAZMATICHESKIKH I IADERNYKH RIBOSOM PECHENI KRYSY. Biokhimiia. 1963 Nov-Dec;28:1047–1052. [PubMed] [Google Scholar]
  9. FLAMM W. G., BIRNSTIEL M. L. THE NUCLEAR SYNTHESIS OF RIBOSOMES IN CELL CULTURES. Biochim Biophys Acta. 1964 May 18;87:101–110. doi: 10.1016/0926-6550(64)90051-9. [DOI] [PubMed] [Google Scholar]
  10. Frenster J. H., Allfrey V. G., Mirsky A. E. METABOLISM AND MORPHOLOGY OF RIBONUCLEOPROTEIN PARTICLES FROM THE CELL NUCLEUS OF LYMPHOCYTES. Proc Natl Acad Sci U S A. 1960 Apr;46(4):432–444. doi: 10.1073/pnas.46.4.432. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. GEORGIEV G. P., CHENTSOV Iu S. [On ultrastructure of the nucleus on the basis of electron microscopy of isolated nuclei subjected to salt extracts]. Biofizika. 1963;8:50–57. [PubMed] [Google Scholar]
  12. HOWELL R. R., LOEB J. N., TOMKINS G. M. CHARACTERIZATION OF RIBOSOMAL AGGREGATES ISOLATED FROM LIVER. Proc Natl Acad Sci U S A. 1964 Nov;52:1241–1248. doi: 10.1073/pnas.52.5.1241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lawford G. R., Langford P., Schachter H. The inhibition of rat liver polyribosome breakdown in the presence of liver supernatant. J Biol Chem. 1966 Apr 25;241(8):1835–1839. [PubMed] [Google Scholar]
  14. MARINOZZI V. CYTOCHIMIE ULTRASTRUCTURALE DU NUCL'EOLE--RNA ET PROT'EINES INTRANUCL'EOLAIRES. J Ultrastruct Res. 1964 Jun;10:433–456. doi: 10.1016/s0022-5320(64)80021-6. [DOI] [PubMed] [Google Scholar]
  15. McCarty K. S., Parsons J. T., Carter W. A., Laszlo J. Protein-synthetic capacities of liver nuclear subfractions. J Biol Chem. 1966 Dec 10;241(23):5489–5499. [PubMed] [Google Scholar]
  16. Muramatsu M., Hodnett J. L., Steele W. J., Busch H. Synthesis of 28-S RNA in the nucleolus. Biochim Biophys Acta. 1966 Jul 20;123(1):116–125. doi: 10.1016/0005-2787(66)90164-x. [DOI] [PubMed] [Google Scholar]
  17. Narayan K. S., Steele W. J., Smetana K., Busch H. Ultrastructural aspects of the ribonucleo-protein network in nuclei of Walker tumor and rat liver. Exp Cell Res. 1967 Apr;46(1):65–77. doi: 10.1016/0014-4827(67)90409-0. [DOI] [PubMed] [Google Scholar]
  18. Penman S. RNA metabolism in the HeLa cell nucleus. J Mol Biol. 1966 May;17(1):117–130. doi: 10.1016/s0022-2836(66)80098-0. [DOI] [PubMed] [Google Scholar]
  19. RENDI R. In vitro incorporation of labeled amino acids into nuclei isolated from rat liver. Exp Cell Res. 1960 Apr;19:489–498. doi: 10.1016/0014-4827(60)90058-6. [DOI] [PubMed] [Google Scholar]
  20. ROTH J. S. Ribonuclease. V. Studies on the properties and distribution of ribonuclease inhibitor in the rat. Biochim Biophys Acta. 1956 Jul;21(1):34–43. doi: 10.1016/0006-3002(56)90091-9. [DOI] [PubMed] [Google Scholar]
  21. ROTH J. S. Ribonuclease. VII. Partial purification and characterization of a ribonuclease inhibitor in rat liver supernatant fraction. J Biol Chem. 1958 Apr;231(2):1085–1095. [PubMed] [Google Scholar]
  22. Robbins E., Borun T. W. The cytoplasmic synthesis of histones in hela cells and its temporal relationship to DNA replication. Proc Natl Acad Sci U S A. 1967 Feb;57(2):409–416. doi: 10.1073/pnas.57.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sadowski P. D., Steiner J. W. Electron microscopic and biochemical characteristics of nuclei and nucleoli isolated from rat liver. J Cell Biol. 1968 Apr;37(1):147–161. doi: 10.1083/jcb.37.1.147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Shin D. H., Moldave K. The stimulation of RNA synthesis by ribosomes in vitro. Biochem Biophys Res Commun. 1966 Jan 24;22(2):232–236. doi: 10.1016/0006-291x(66)90438-4. [DOI] [PubMed] [Google Scholar]
  25. Siebert G., Villalobos J., Jr, Ro T. S., Steele W. J., Lindenmayer G., Adams H., Busch H. Enzymatic studies on isolated nucleoli of rat liver. J Biol Chem. 1966 Jan 10;241(1):71–78. [PubMed] [Google Scholar]
  26. Singer M. F., Leder P. Messenger RNA: an evaluation. Annu Rev Biochem. 1966;35:195–230. doi: 10.1146/annurev.bi.35.070166.001211. [DOI] [PubMed] [Google Scholar]
  27. TRAUB A., KAUFMANN E., GINZBURG-TIETZ Y. STUDIES ON NUCLEAR RIBOSOMES. I. ASSOCIATION OF DPN-PYROPHOSPHORYLASE WITH NUCLEAR RIBOSOMES IN NORMAL AND NEOPLASTIC TISSUES. Exp Cell Res. 1964 Apr;34:371–383. doi: 10.1016/0014-4827(64)90372-6. [DOI] [PubMed] [Google Scholar]
  28. WATSON J. D. Involvement of RNA in the synthesis of proteins. Science. 1963 Apr 5;140(3562):17–26. doi: 10.1126/science.140.3562.17. [DOI] [PubMed] [Google Scholar]
  29. WETTSTEIN F. O., STAEHELIN T., NOLL H. Ribosomal aggregate engaged in protein synthesis: characterization of the ergosome. Nature. 1963 Feb 2;197:430–435. doi: 10.1038/197430a0. [DOI] [PubMed] [Google Scholar]
  30. Wilson S. H., Hoagland M. B. Studies on the physiology of rat liver polyribosomes: quantition and intracellular distribution of ribosomes. Proc Natl Acad Sci U S A. 1965 Aug;54(2):600–607. doi: 10.1073/pnas.54.2.600. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES