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. 1971 Aug;68(8):1881–1885. doi: 10.1073/pnas.68.8.1881

Isolation of a 5S RNA-Protein Complex from Mammalian Ribosomes

Günter Blobel 1
PMCID: PMC389313  PMID: 5001943

Abstract

5S RNA is removed from the large ribosomal subunit of both rat liver and rabbit reticulocyte ribosomes by treatment with EDTA to remove magnesium ions. The RNA is removed not as a naked molecule, but in association with a single ribosomal protein (molecular weight about 35,000) as a ribonucleoprotein, sedimenting at about 7 S. Conditions for the exclusive removal of the 5S RNA-protein complex have been established.

Both ribosomal subunits undergo distinct changes in their sedimentation rate, concomitant with the loss of their biological activity, when sedimented into sucrose gradients containing high concentrations of monovalent ions and low concentrations of magnesium. Under these conditions the large subunit loses 5S RNA, the 5S RNA-associated protein, and several more proteins.

Keywords: rat liver, rabbit reticulocytes

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Selected References

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

  1. Blobel G. Release, identification, and isolation of messenger RNA from mammalian ribosomes. Proc Natl Acad Sci U S A. 1971 Apr;68(4):832–835. doi: 10.1073/pnas.68.4.832. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blobel G., Sabatini D. Dissociation of mammalian polyribosomes into subunits by puromycin. Proc Natl Acad Sci U S A. 1971 Feb;68(2):390–394. doi: 10.1073/pnas.68.2.390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brown D. D., Littna E. Synthesis and accumulation of low molecular weight RNA during embryogenesis of Xenopus laevis. J Mol Biol. 1966 Sep;20(1):95–112. doi: 10.1016/0022-2836(66)90120-3. [DOI] [PubMed] [Google Scholar]
  4. Brownlee G. G., Sanger F., Barrell B. G. Nucleotide sequence of 5S-ribosomal RNA from Escherichia coli. Nature. 1967 Aug 12;215(5102):735–736. doi: 10.1038/215735a0. [DOI] [PubMed] [Google Scholar]
  5. COMB D. G., SARKAR N., DEVALLET J., PINZINO C. J. PROPERTIES OF TRANSFER-LIKE RNA ASSOCIATED WITH RIBOSOMES. J Mol Biol. 1965 Jun;12:509–513. doi: 10.1016/s0022-2836(65)80273-x. [DOI] [PubMed] [Google Scholar]
  6. Comb D. G., Sarkar N. The binding of 5s ribosomal ribonucleic acid to ribosomal subunits. J Mol Biol. 1967 Apr 28;25(2):317–330. doi: 10.1016/0022-2836(67)90144-1. [DOI] [PubMed] [Google Scholar]
  7. Dahlberg A. E., Dingman C. W., Peacock A. C. Electrophoretic characterization of bacterial polyribosomes in agarose-acrylamide composite gels. J Mol Biol. 1969 Apr 14;41(1):139–147. doi: 10.1016/0022-2836(69)90131-4. [DOI] [PubMed] [Google Scholar]
  8. Dingman C. W., Peacock A. C. Analytical studies on nuclear ribonucleic acid using polyacrylamide gel electrophoresis. Biochemistry. 1968 Feb;7(2):659–668. doi: 10.1021/bi00842a022. [DOI] [PubMed] [Google Scholar]
  9. Forget B. G., Weissman S. M. Nucleotide sequence of KB cell 5S RNA. Science. 1967 Dec 29;158(3809):1695–1699. doi: 10.1126/science.158.3809.1695. [DOI] [PubMed] [Google Scholar]
  10. Galibert F., Larsen C. J., Lelong J. C., Boiron M. RNA of low molecular weight in ribosomes of mammalian cells. Nature. 1965 Sep 4;207(5001):1039–1041. doi: 10.1038/2071039a0. [DOI] [PubMed] [Google Scholar]
  11. Knight E., Jr, Darnell J. E. Distribution of 5 s RNA in HeLa cells. J Mol Biol. 1967 Sep 28;28(3):491–502. doi: 10.1016/s0022-2836(67)80099-8. [DOI] [PubMed] [Google Scholar]
  12. Lebleu B., Marbaix G., Huez G., Temmerman J., Burny A., Chantrenne H. Characterization of the messenger ribonucleoprotein released from reticulocyte polyribosomes by EDTA treatment. Eur J Biochem. 1971 Mar 11;19(2):264–269. doi: 10.1111/j.1432-1033.1971.tb01313.x. [DOI] [PubMed] [Google Scholar]
  13. MARCOT-QUEIROZ J., JULIEN J., ROSSET R., MONIER R. LES ACIDES RIBONUCL'EIQUES DES RIBOSOMES DE LA LEVURE. Bull Soc Chim Biol (Paris) 1965;47:183–194. [PubMed] [Google Scholar]
  14. MARTIN R. G., AMES B. N. A method for determining the sedimentation behavior of enzymes: application to protein mixtures. J Biol Chem. 1961 May;236:1372–1379. [PubMed] [Google Scholar]
  15. Morell P., Marmur J. Association of 5S ribonucleic acid to 50S ribosomal subunits of Escherichia coli and Bacillus subtilis. Biochemistry. 1968 Mar;7(3):1141–1152. doi: 10.1021/bi00843a035. [DOI] [PubMed] [Google Scholar]
  16. ROSSET R., MONIER R., JULIEN J. LES RIBOSOMES D'ESCHERICHIA COLI. I. MISE EN 'EVIDENCE D'UN RNA RIBOSOMIQUE DE FAIBLE POIDS MOL'ECULAIRE. Bull Soc Chim Biol (Paris) 1964;46:87–109. [PubMed] [Google Scholar]
  17. Sarkar N., Comb D. G. Studies on the attachment and release of 5 s ribosomal RNA from the large ribosomal subunit. J Mol Biol. 1969 Jan 14;39(1):31–44. doi: 10.1016/0022-2836(69)90331-3. [DOI] [PubMed] [Google Scholar]
  18. Siddiqui M. A., Hosokawa K. Role of 5S ribosomal RNA in polypeptide synthesis. II. Dissociation of 5S ribosomal RNA from 50S ribosomes in Escherichia coli. Biochem Biophys Res Commun. 1968 Jul 11;32(1):1–8. doi: 10.1016/0006-291x(68)90417-8. [DOI] [PubMed] [Google Scholar]
  19. Solymosy F., Fedorcsák I., Gulyás A., Farkas G. L., Ehrenberg L. A new method based on the use of diethyl pyrocarbonate as a nuclease inhibitor for the extraction of undegraded nucleic acid from plant tissues. Eur J Biochem. 1968 Sep 24;5(4):520–527. doi: 10.1111/j.1432-1033.1968.tb00401.x. [DOI] [PubMed] [Google Scholar]
  20. TASHIRO Y., SIEKEVITZ P. ULTRACENTRIFUGAL STUDIES ON THE DISSOCIATION OF HEPATIC RIBOSOMES. J Mol Biol. 1965 Feb;11:149–165. doi: 10.1016/s0022-2836(65)80047-x. [DOI] [PubMed] [Google Scholar]
  21. Zehavi-Willner T. The release of 5S RNA from reticulocyte ribosomes. Biochem Biophys Res Commun. 1970 Apr 8;39(1):161–169. doi: 10.1016/0006-291x(70)90772-2. [DOI] [PubMed] [Google Scholar]

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