RNA-protein cross-linking in Escherichia coli 30S ribosomal subunits: Precise localisation of the nucleotide in 16S RNA which is coupled to protein S7 by ultraviolet irradiation

Christian Zwieb, Richard Brimacornbe

Research output: Contribution to journalArticle

63 Scopus citations

Abstract

It is well established that when E. coil 30s ribosomal subunits are irradiated with ultraviolet light under mild conditions a specific cross-link is formed between protein S7 and the 16s RNA. Methodology is presented for the analysis of the single nucleotide residue concerned in this cross-link. Firstly, the identity of the ribonuclease T1 octanucleotide attached to S7 is confirmed by a new method, which involves isolation and analysis of S7-polynucleotide complexes containing 30 - 40 nucleotides. Secondly, the isolated S7-octanucleotide complex is digested successively with ribonuclease A, proteinase K and ribonuclease T2, and the nucleotides liberated are identified. The results show unambiguously that uridine residue number 1239 in the 16s RNA sequence is cross-linked to protein S7.

Original languageEnglish (US)
Pages (from-to)1775-1790
Number of pages16
JournalNucleic acids research
Volume6
Issue number5
DOIs
StatePublished - Dec 1 1979
Externally publishedYes

ASJC Scopus subject areas

  • Genetics

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