Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1967 Aug 1;34(2):505–512. doi: 10.1083/jcb.34.2.505

OCCURRENCE, ISOLATION, AND CHARACTERIZATION OF POLYRIBOSOMES IN YEAST

Leon Marcus 1, H Ris 1, H O Halvorson 1, R K Bretthauer 1, R M Bock 1
PMCID: PMC2107312  PMID: 6035641

Abstract

This report details the procedural requirements for preparing cell-free extracts of yeast rich in polyribosomes. This enabled us to demonstrate the occurrence of polyribosomes in yeast, to show their role in protein synthesis, and to devise methods for their resolution and isolation. When certain precautions are met (the use of log phase cells, rapidly halting cell growth, gentle methods of disruption, sedimentation through exponential density gradients, etc.), individual polyribosome size classes ranging up to the heptosome can be fractionated and separated from their nearest neighbors. Larger size classes are resolved partially among themselves, free of smaller polyribosomes. This was confirmed by extensive electron micrographic studies of material from the various fractions obtained upon density gradient centrifugation of yeast extracts. Modifications of the gradients and procedure should allow fractionation and isolation of the larger polyribosomes, including those containing polycistronic messages. Yeast polyribosomes are disaggregated to single ribosomes by longer term grinding, cell disruption by the French pressure cell, the Hughes press, or by incubation with dilute RNAse. Yeast polyribosomes are active in the incorporation of amino acids into polypeptide; the single ribosomes exhibit only slight activity. The latter activity is probably due to the presence of a small fraction of monosomes still containing mRNA. Poly-U stimulates amino acid incorporation only in the single ribosomes.

Full Text

The Full Text of this article is available as a PDF (668.0 KB).

Selected References

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

  1. BRETTHAUER R. K., MARCUS L., CHALOUPKA J., HALVORSON H. O., BOCK R. M. AMINO ACID INCORPORATION INTO PROTEIN BY CELL-FREE EXTRACTS OF YEAST. Biochemistry. 1963 Sep-Oct;2:1079–1084. doi: 10.1021/bi00905a029. [DOI] [PubMed] [Google Scholar]
  2. GIERER A. Function of aggregated reticulocyte ribosomes in protein synthesis. J Mol Biol. 1963 Feb;6:148–157. doi: 10.1016/s0022-2836(63)80131-x. [DOI] [PubMed] [Google Scholar]
  3. GILBERT W. Polypeptide synthesis in Escherichia coli. I. Ribosomes and the active complex. J Mol Biol. 1963 May;6:374–388. doi: 10.1016/s0022-2836(63)80050-9. [DOI] [PubMed] [Google Scholar]
  4. KITAZUME Y., YCAS M. THE CALCULATED COMPOSITION OF NEWLY SYNTHESIZED YEAST RIBONUCLEIC ACID. Biochim Biophys Acta. 1963 Nov 22;76:391–400. [PubMed] [Google Scholar]
  5. MARCUS L., BRETTHAUER R. K., BOCK R. M., HALVORSON H. O. THE EFFECT OF POLY U SIZE ON THE INCORPORATION OF PHENYLALANINE IN THE CELL-FREE YEAST SYSTEM. Proc Natl Acad Sci U S A. 1963 Oct;50:782–789. doi: 10.1073/pnas.50.4.782. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. MARCUS L., BRETTHAUER R., HALVORSON H., BOCK R. POLYSOMES FROM YEAST: DISTRIBUTION OF MESSENGER RNA AND CAPACITY TO SUPPORT PROTEIN SYNTHESIS IN VITRO. Science. 1965 Feb 5;147(3658):615–617. doi: 10.1126/science.147.3658.615. [DOI] [PubMed] [Google Scholar]
  7. Penman S., Scherrer K., Becker Y., Darnell J. E. POLYRIBOSOMES IN NORMAL AND POLIOVIRUS-INFECTED HELA CELLS AND THEIR RELATIONSHIP TO MESSENGER-RNA. Proc Natl Acad Sci U S A. 1963 May;49(5):654–662. doi: 10.1073/pnas.49.5.654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. RISEBROUGH R. W., TISSIERES A., WATSON J. D. Messenger-RNA attachment to active ribosomes. Proc Natl Acad Sci U S A. 1962 Mar 15;48:430–436. doi: 10.1073/pnas.48.3.430. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. SLAYTER H. S., WARNER J. R., RICH A., HALL C. E. THE VISUALIZATION OF POLYRIBOSOMAL STRUCTURE. J Mol Biol. 1963 Dec;7:652–657. doi: 10.1016/s0022-2836(63)80112-6. [DOI] [PubMed] [Google Scholar]
  10. STAEHELIN T., BRINTON C. C., WETTSTEIN F. O., NOLL H. STRUCTURE AND FUNCTION OF E. COLI ERGOSOMES. Nature. 1963 Aug 31;199:865–870. doi: 10.1038/199865a0. [DOI] [PubMed] [Google Scholar]
  11. WARNER J. R., KNOPF P. M., RICH A. A multiple ribosomal structure in protein synthesis. Proc Natl Acad Sci U S A. 1963 Jan 15;49:122–129. doi: 10.1073/pnas.49.1.122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. YOUNG R. J., KIHARA H. K., HALVORSON H. O. Studies on the kinetics of protein synthesis in yeast. Proc Natl Acad Sci U S A. 1961 Sep 15;47:1415–1423. doi: 10.1073/pnas.47.9.1415. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES