Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1981 Nov;78(11):6652–6656. doi: 10.1073/pnas.78.11.6652

Arrangement of the subunits in the ribosome of Escherichia coli: demonstration by immunoelectron microscopy.

B Kastner, M Stöffler-Meilicke, G Stöffler
PMCID: PMC349107  PMID: 7031657

Abstract

The three-dimensional locations of Escherichia coli ribosomal proteins S13, L1, and L7/L12 on the surface of ribosomal subunits and 70S monomeric ribosomes were determined by electron microscopy of antibody-labeled ribosomal particles. A new approach to orient the subunits within 70S ribosomes was developed that used 30S.70S.50S triples that were prepared by simultaneous combination with one antibody directed against a 30S protein and another directed against a 50S protein. Electron microscope studies of triples obtained with the antibody combinations anti-S13/anti-L1 and anti-S13/anti-L7/L12 showed that, in 70S monomeric ribosomes, the head of the 30S subunit is proximate to protein L1 and the peptidyl transferase center but far from the rod-like appendage containing proteins L7 and L12.

Full text

PDF
6652

Images in this article

6653Image
on p.6653
6653Image
on p.6653
6654Image
on p.6654
6655Image
on p.6655
6655Image
on p.6655

Selected References

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

  1. Boublik M., Hellmann W., Roth H. E. Localization of ribosomal proteins L7L12 in the 50 S subunit of Escherichia coli Ribosomes by electron microscopy. J Mol Biol. 1976 Nov 15;107(4):479–490. doi: 10.1016/s0022-2836(76)80079-4. [DOI] [PubMed] [Google Scholar]
  2. Dabbs E. R., Ehrlich R., Hasenbank R., Schroeter B. H., Stöffler-Meilicke M., Stöffler G. Mutants of Escherichia coli lacking ribosomal protein L1. J Mol Biol. 1981 Jul 15;149(4):553–578. doi: 10.1016/0022-2836(81)90347-8. [DOI] [PubMed] [Google Scholar]
  3. Keren-Zur M., Boublik M., Ofengand J. Localization of the decoding region on the 30S Escherichia coli ribosomal subunit by affinity immunoelectron microscopy. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1054–1058. doi: 10.1073/pnas.76.3.1054. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Korn A. P. Dark field electron microscopy of the 30S subunit of the E. coli ribosome. Ultramicroscopy. 1980;5(4):513–520. doi: 10.1016/s0304-3991(80)80008-8. [DOI] [PubMed] [Google Scholar]
  5. Lake J. A., Kahan L. Ribosomal proteins S5, S11, S13 and S19 localized by electron microscopy of antibody-labeled subunits. J Mol Biol. 1975 Dec 25;99(4):631–644. doi: 10.1016/s0022-2836(75)80177-x. [DOI] [PubMed] [Google Scholar]
  6. Lake J. A. Ribosome structure determined by electron microscopy of Escherichia coli small subunits, large subunits and monomeric ribosomes. J Mol Biol. 1976 Jul 25;105(1):131–139. doi: 10.1016/0022-2836(76)90200-x. [DOI] [PubMed] [Google Scholar]
  7. Leonard K. R., Lake J. A. Ribosome structure: hand determination by electron microscopy of 30 S subunits. J Mol Biol. 1979 Mar 25;129(1):155–163. doi: 10.1016/0022-2836(79)90066-4. [DOI] [PubMed] [Google Scholar]
  8. Lubin M. Observations on the shape of the 50S ribosomal subunit. Proc Natl Acad Sci U S A. 1968 Dec;61(4):1454–1461. doi: 10.1073/pnas.61.4.1454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Miyazawa Y., Hosoi J., Imamura T., Mochizuki A. Mutual relations between typical images of negatively stained ribosomal monomer and large subunit particles from E. coli: an electron microscopic study with tilting technique. J Electron Microsc (Tokyo) 1978;27(3):189–197. [PubMed] [Google Scholar]
  10. Noll M., Hapke B., Schreier M. H., Noll H. Structural dynamics of bacterial ribosomes. I. Characterization of vacant couples and their relation to complexed ribosomes. J Mol Biol. 1973 Apr 5;75(2):281–294. doi: 10.1016/0022-2836(73)90021-1. [DOI] [PubMed] [Google Scholar]
  11. Olson H. M., Grant P. G., Glitz D. G., Cooperman B. S. Immunoelectron microscopic localization of the site of photo-induced affinity labeling of the small ribosomal subunit with puromycin. Proc Natl Acad Sci U S A. 1980 Feb;77(2):890–894. doi: 10.1073/pnas.77.2.890. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Spiess E. Structure of the 50S ribosomal subunit from Escherichia coli. Investigation of the intact subunit and core particles by electron microscopy and analogue image processing. Eur J Cell Biol. 1979 Jun;19(2):120–130. [PubMed] [Google Scholar]
  13. Strycharz W. A., Nomura M., Lake J. A. Ribosomal proteins L7/L12 localized at a single region of the large subunit by immune electron microscopy. J Mol Biol. 1978 Dec 5;126(2):123–140. doi: 10.1016/0022-2836(78)90355-8. [DOI] [PubMed] [Google Scholar]
  14. Stöffler G., Hasenbank R., Lütgehaus M., Maschler R., Morrison C. A., Zeichhardt H., Garrett R. A. The accessibility of proteins of the Escherichia coli 30S ribosomal subunit to antibody binding. Mol Gen Genet. 1973 Dec 20;127(2):89–110. doi: 10.1007/BF00333659. [DOI] [PubMed] [Google Scholar]
  15. Stöffler G., Wittmann H. G. Ribosomal proteins. XXV. Immunological studies on Escherichia coli ribosomal proteins. J Mol Biol. 1971 Dec 14;62(2):407–409. doi: 10.1016/0022-2836(71)90436-0. [DOI] [PubMed] [Google Scholar]
  16. Stöffler G., Wittmann H. G. Sequence differences of Escherichia coli 30S ribosomal proteins as determined by immunochemical methods. Proc Natl Acad Sci U S A. 1971 Sep;68(9):2283–2287. doi: 10.1073/pnas.68.9.2283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Tischendorf G. W., Stöffler G. Localization of Escherichia coli ribosomal protein S4 on the surface of the 30S ribosomal subunit by immuno electron microscopy. I. Distribution of antibody-binding sites as obtained with immunoglobulins and monovalent antibody fragments from various S4-specific antisera. Mol Gen Genet. 1975 Dec 30;142(3):193–208. doi: 10.1007/BF00425645. [DOI] [PubMed] [Google Scholar]
  18. Tischendorf G. W., Zeichhardt H., Stöffler G. Architecture of the Escherichia coli ribosome as determined by immune electron microscopy. Proc Natl Acad Sci U S A. 1975 Dec;72(12):4820–4824. doi: 10.1073/pnas.72.12.4820. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tischendorf G. W., Zeichhardt H., Stöffler G. Determination of the location of proteins L14, L17, L18, L19, L22, L23 on the surface of the 5oS ribosomal subunit of Escherichia coli by immune electron microscopy. Mol Gen Genet. 1974;134(3):187–208. doi: 10.1007/BF00267715. [DOI] [PubMed] [Google Scholar]
  20. Tischendorf G. W., Zeichhardt H., Stöffler G. Location of proteins S5, S13 and S14 on the surface of the 3oS ribosomal subunit from Escherichia coli as determined by immune electron microscopy. Mol Gen Genet. 1974;134(3):209–223. doi: 10.1007/BF00267716. [DOI] [PubMed] [Google Scholar]
  21. Vasiliev V. D. Morphology of the ribosomal 30S subparticle according to electron microscopic data. Acta Biol Med Ger. 1974;33(5-6):779–793. [PubMed] [Google Scholar]
  22. Wabl M. R. Electron microscopic localization of two proteins on the surface of the 50 S ribosomal subunit of Escherichia coli using specific antibody markers. J Mol Biol. 1974 Apr 5;84(2):241–247. doi: 10.1016/0022-2836(74)90582-8. [DOI] [PubMed] [Google Scholar]
  23. Zubke W., Stadler H., Ehrlich R., Stöffler G., Wittmann H. G., Apirion D. Improved electrophoretic and immunochemical techniques for the identification and characterization of mutant proteins, applied to ribosomal protein S8 in Escherichia coli mutants. Mol Gen Genet. 1977 Dec 30;158(2):129–139. doi: 10.1007/BF00268305. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES