The localization of multiple sites on 16S RNA which are cross-linked to proteins S7 and S8 in Escherichia coli 30S ribosomal subunits by treatment with 2-iminothiolane.
RNA-protein cross-links were introduced into E. coli 30S ribosomal subunits by reaction with 2-iminothiolane followed by a mild ultraviolet irradiation treatment. After removal of non-reacted protein and partial nuclease digestion of the cross-linked 16S RNA-protein moiety, a number of individual cross-linked complexes could be isolated and the sites of attachment of the proteins to the RNA determined. Protein S8 was cross-linked to the RNA at three different positions, within oligo-nucleotides encompassing positions 629-633, 651-654, and (tentatively) 593-597 in the 16S sequence. Protein S7 was cross-linked within two oligonucleotides encompassing positions 1238-1240, and 1377-1378. In addition, a site at position 723-724 was observed, cross-linked to protein S19, S20 or S21.
D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000032:analysis*; D007096:Imidoesters; D008958:Models, Molecular; D008970:Molecular Weight; D009690:Nucleic Acid Conformation; D011487:Protein Conformation; D012335:RNA, Ribosomal / Q000032:analysis*; D012269:Ribosomal Proteins / Q000032:analysis*; D012270:Ribosomes / Q000032:analysis / Q000648:ultrastructure
D003432:Cross-Linking Reagents; D007096:Imidoesters; D012335:RNA, Ribosomal; D012269:Ribosomal Proteins; C037442:ribosomal protein S7; C030254:ribosomal protein S8; C005905:methyl 4-mercaptobutyrimidate