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Nucleic Acids Research, 1986, Vol. 14, No. 3 1171-1186
© 1986

Articles

RNA-protein cross-linking in Escherichia coli ribosomal subunits: localization of sites on 16S RNA which are cross-linked to proteins S17 and S21 by treatment with 2-iminothiolane

Apostolos Kyriatsoulis*,, Peter Maly+,, Barbara Greuer, Richard Brimacombe§, Georg Stöffler1, Ronald Frank2 and Helmut Blöcker2

1Institut fur Mikrobiologie, Medizinische Fakultät, Universitat Innsbruck Fritz-Pregl-Str. 3, A-6020, Innsbruck, Austria 2Gesellschaft für Biotechnologische Forschung Abteilung DNA-Synthese D-3000 Braunschweig-Stöckheim, FRG Max-Planck-Institut für Molekulare Genetik, Abteilung Wittmann D-1000 Berlin-Dahlem, FRG

§To whom correspondence should be addressed

Received November 29, 1985. Accepted January 14, 1986.

Treatment of E. coli ribosomal subunits with 2-iminothiolane coupled with mild ultraviolet irradiation leads to the formation of a large number of RNA-protein cross-links. In the case of the 30S subunit, a number of sites on 16S RNA that are cross-linked to proteins S7 and S8 by this procedure have aLready been identified (see ref. 6). Here, by using new or modified techniques for the partial digestion of the RNA and the subsequent isolation of the cross-linked RNA-protein complexes, three new iminothiolane cross-links have been localized: Protein S17 is cross-linked to the 16S RNA within an oligonucleotide encompassing positions 629–633, and protein S21 is cross-linked to two sites within oligonucleotides encompassing positions 723–724 and positions 1531–1542 (the 3'-end of the 16S RNA).

*Present addresses:I. Medizinische Klinik der Johannes Gutenberg Universität, Langenwitstr. 1, D-6500 Mainz, FRG

+Present addresses:Biochemisches Institut der Universität Zürich, Winterthurerstr. 190, CH-8057 Zürich, Switzerland


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