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PMID: 349502 Published · ppublish English Journal Article

RNA-protein cross-linking in Eschericia coli 30S ribosomal subunits: a method for the direct analysis of the RNA regions involved in the cross-links.

Nucleic acids research ·Vol. 5 ·No. 4 ·1978-04-00 ·Pages 1189-206

Zwieb C, Brimacombe R

Abstract

A prerequisite for topographical studies on ribosomal subunits involving RNA-protein cross-linking is that the cross-linking sites on the RNA should be determined. Methodology is presented which offers a solution to this problem, using as a test system 30S subunits in which protein S7 has been cross-linked to the 16S RNA by ultraviolet irradiation. The method is based on a gel separation system in the presence of a non-ionic detergent. When a ribonucleoprotein fragment containing RNA-protein cross-links is applied to this system, non-cross-linked protein is removed, and simultaneously the cross-linked RNA-protein complex is separated from non-cross-linked RNA. Oligonucleotide analysis of the S7-RNA complex isolated in this manner showed it to consist of a region of RNA from sections P-A of the 16S RNA. A single characteristic oligonucleotide was absent from this region, and it was tentatively concluded that this missing oligonucleotide contains the actual site of cross-linking.

MeSH Terms
Bacterial Proteins/radiation effects Binding Sites Escherichia coli Molecular Weight Oligoribonucleotides/analysis Protein Binding RNA, Bacterial/radiation effects Ribosomes/radiation effects
Chemicals
Bacterial Proteins Oligoribonucleotides RNA, Bacterial
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Zwieb C
Brimacombe R
References (19)
19 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/5.4.1189
Published
1978-04-00
Pages
1189-206
Language
English
Region
England
NLM ID
0411011
PMCID
PMC342070
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/5.4.1189 PMC Full Text View journal details
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