RNA-protein cross-linking in Escherichia coli 30S ribosomal subunits: determination of sites on 16S RNA that are cross-linked to proteins S3, S4, S5, S7, S8, S9, S11, S13, S19 and S21 by treatment with methyl p-azidophenyl acetimidate

RNA-protein cross-links were introduced into E. coli 30S ribosomal subunits by treatment with methyl p-azidophenyl acetimidate. After partial nuclease digestion of the RNA moiety, a number of cross-linked RNA-protein complexes were isolated by a new three-step procedure. Protein and RNA analysis of...

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Bibliographic Details
Published in:Nucleic acids research 1987-04, Vol.15 (8), p.3221-3240
Main Authors: OSSWALD, M, GREUER, B, BRIMACOMBE, R, STÖFFLER, G, BÄUMERT, H, FASOLD, H
Format: Article
Language:English
Subjects:
Bacterial Proteins - analysis
Bacteriology
Base Sequence
Biogenesis of cell structures, supramolecular organization
Biological and medical sciences
Cross-Linking Reagents
Escherichia coli
Escherichia coli - analysis
Fundamental and applied biological sciences. Psychology
Imidoesters
Microbiology
Nucleic Acid Conformation
Ribosomal Proteins - analysis
RNA, Bacterial - analysis
RNA, Ribosomal - analysis
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Summary:RNA-protein cross-links were introduced into E. coli 30S ribosomal subunits by treatment with methyl p-azidophenyl acetimidate. After partial nuclease digestion of the RNA moiety, a number of cross-linked RNA-protein complexes were isolated by a new three-step procedure. Protein and RNA analysis of the individual complexes gave the following results: Proteins S3, S4, S5 and S8 are cross-linked to the 5'-terminal tetranucleotide of 16S RNA. S5 is also cross-linked to the 16S RNA within an oligonucleotide encompassing positions 559-561. Proteins S11, S9, S19 and S7 are cross-linked to 16S RNA within oligonucleotides encompassing positions 702-705, 1130-1131, 1223-1231 and 1238-1240, respectively. Protein S13 is cross-linked to an oligonucleotide encompassing positions 1337-1338, and is also involved in an anomalous cross-link within positions 189-191. Protein S21 is cross-linked to the 3'-terminal dodecanucleotide of the 16S RNA.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/15.8.3221