Work

Quantitative structural analysis of eukaryotic ribosomal RNA by scanning transmission electron microscopy.

Year: 1985

Type: article

Abstract: The conformation of 28S ribosomal RNA isolated from baby hamster kidney cells was studied by scanning transmission electron microscopy (STEM) and circular dichroic spectroscopy to establish the conditions under which STEM images of unstained freeze-dried rRNA are a meaningful representation of the conformation of rRNA in solution. We have determined the conformation of 28S rRNA under various buffer conditions, the molecular mass, the mass distribution, and the number of polynucleotide strands within the individual molecules, and the apparent radii of gyration. The 28S rRNA molecule is highly extended in water and becomes compact with increasing ionic strength. However, even in the "reconstitution buffer" (30 mM Tris/HCl/20 mM MgCl2/360 mM KCl, pH 7.6) the compactness does not reach a state in which the rRNA molecule appears structurally similar to the 60S ribosomal subunit. Our approach has a broad application in high-resolution structural studies of nucleic acids and nucleic acid-protein interactions.

Source: Proceedings of the National Academy of Sciences

DOI: 10.1073/pnas.82.17.5598

Authors: G T Oostergetel, J S Wall, J F Hainfeld, M Boublik

Institution: Brookhaven National Laboratory

Language: English

Topic: RNA and protein synthesis mechanisms

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funders: National Institutes of Health, National Center for Research Resources

Award: G869871382