Crosslinking of ribosomal proteins S4, S5, S7, S8, S11, S12 and S18 to domains 1 and 2 of 16S rRNA in the Escherichia coli 30S particle.
RNA-protein crosslinks were introduced into Escherichia coli 30S ribosomal subunits by treatment with 1-ethyl-3(3-dimethylaminopropyl)carbodiimide (EDC). Complexes of 16S rRNA cross-linked to 30S ribosomal proteins were isolated and hybridized with a series of single-stranded bacteriophage M13-rDNA probes. These probes, each carrying an inserted rDNA fragment, were used to select contiguous 16S rRNA sections covering all of domain 1 and the major part of domain 2 (starting at the 5'-P terminus and ending at nucleotide 869) and the proteins covalently linked to each of these sections were identified by 2-dimensional polyacrylamide gel electrophoresis. This procedure identified proteins S4, S5, S7, S8, S11, S12, and S18 as the species most efficiently crosslinked to domains 1 and 2 of 16S rRNA. These results are discussed in the light of current knowledge of the tertiary structure of 16S rRNA in the E. coli 30S ribosomal subunit.
Chiaruttini
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C
Laboratoire de Chimie Cellulaire, Institut de Biologie Physico-chimique, Paris, France.
Milet
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M
Hayes
D H
DH
Expert-Bezancon
A
A
D003432:Cross-Linking Reagents / Q000494:pharmacology; D015342:DNA Probes; D004586:Electrophoresis; D004926:Escherichia coli / Q000378:metabolism*; D005022:Ethyldimethylaminopropyl Carbodiimide; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization; D012335:RNA, Ribosomal / Q000378:metabolism*; D012336:RNA, Ribosomal, 16S / Q000378:metabolism*; D012269:Ribosomal Proteins / Q000378:metabolism*
D003432:Cross-Linking Reagents; D015342:DNA Probes; D012335:RNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D012269:Ribosomal Proteins; C040469:ribosomal protein S30; D005022:Ethyldimethylaminopropyl Carbodiimide