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PMID: 386285 Published · ppublish English Journal Article

Binding of magnesium ions and ethidium bromide: comparison of ribosomes and free ribosomal RNA.

Nucleic acids research ·Vol. 7 ·No. 2 ·1979-09-25 ·Pages 465-80

Elson D, Spitnik-Elson P, Avital S, Abramowitz R

Abstract

Comparative studies of free ribosomal RNA and ribosomes were made with two probes, Mg++ ions and ethidium bromide, which interact with RNA in different ways. Mg++. E. coli 16 S rRNA and 30 S ribosomes were equilibrated with four different buffers. Equilibration required several days at 4 degrees and several hours at 37 degrees. In all buffers ribosomes bound more Mg than free rRNA, the difference sometimes reaching 20--30%. Ribosomes were more resistant than free rRNA to heat denaturation and their denaturation was more highly cooperative. Ribosomes that bound more Mg++ had higher denaturation temperatures. Ethidium bromide. Fluorescence enhancement studies of ethidium intercalation showed the free 16 S rRNA to have 50--80 binding sites per molecule. A large fraction of these sites were present and accessible in the ribosome, but their ethidium-binding constants were reduced by an order of magnitude. In addition, free rRNA contained a small number of very strong binding sites that were virtually absent in the ribosomes.

MeSH Terms
Chemical Phenomena Chemistry Escherichia coli/metabolism Ethidium Kinetics Magnesium/metabolism Nucleic Acid Conformation RNA, Ribosomal/metabolism Ribosomes/metabolism,ultrastructure Spectrometry, Fluorescence Temperature
Chemicals
RNA, Ribosomal Ethidium Magnesium
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Elson D
Spitnik-Elson P
Avital S
Abramowitz R
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38 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/7.2.465
Published
1979-09-25
Pages
465-80
Language
English
Region
England
NLM ID
0411011
PMCID
PMC328029
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/7.2.465 PMC Full Text View journal details
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