Precise localisation of three intra-RNA cross-links in 23S RNA, and aone in 5S RNA, induced by treatment of Escherichia coli 50S ribosomal subunits with bis-(2-chloroethtyl)-methylamine

Wolfgang Stiege, Christian Zwieb, Richard Brimacombe

Research output: Contribution to journalArticlepeer-review

35 Scopus citations

Abstract

Treatment of E. coli 50S ribosomal subsunits with low doses of bis-(2-chloroethyl)-methylamine ("nitrogen mustard") leads to formation of a number of intra-RNA and RNA-protein cross-links. After partial digestion of the cross-linked subunits with cobra venom nuclease, followed by destruction of the protein moiety with proteinase K, complexes containing the intra-RNA cross-links were isolated by two-dimensional gel electrophoresis. The individual complexes were subjected to oligonucleotide analysis,either directly or after a second partial digestion procedure using ribonuclease T1, and the cross-link sites determined. In 235 RNA, the cross-links found were between bases 763 and 1567, 1210 and 1236, 1482 and 1501; in 5S RNA, base 69 was cross-linked to base 107. The significance of these cross-links in relation to the three-dimensional organization of the ribosomal RNA is discussed.

Original languageEnglish (US)
Pages (from-to)7211-7229
Number of pages19
JournalNucleic acids research
Volume10
Issue number22
DOIs
StatePublished - Nov 25 1982
Externally publishedYes

ASJC Scopus subject areas

  • Genetics

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