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Nucleic Acids Research, 1983, Vol. 11, No. 5 1419-1437
© 1983

MOLECULAR BIOLOGY

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

Iwona Wower and Richard Brimacombe

Max-Planck-Institut f{euro}r Molekulare Genetik Abteilung Wittmann, Berlin-Dahlem, FRG

Received December 6, 1982. Revised February 1, 1983. Accepted February 1, 1983.

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.


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