Localization of a series of RNA-protein cross-link sites in the 23S and 5S ribosomal RNA from Escherichia coli, induced by treatment of 50S subunits with three different bifunctional reagents.
50S ribosomal subunits were reacted with bis-(2-chloroethyl)methylamine, 2-iminothiolane or methyl p-azidophenyl acetimidate, and RNA-protein cross-link sites on the RNA were localised using our published procedures. The degree of precision with which these sites could be determined was variable, depending on the particular protein or RNA region concerned. The following positions in the 23S RNA were identified as encompassing the individual cross-link sites (numbered from the 5'-end, with asterisks denoting sites previously reported): L1, 1864-67, 1876-78, 2119-33, 2163-72*, L2, 1819-20*; L3, 2832-34; L4, 320-25*; 613-17*; L5, 2307; L6, 2473-81*; L9, 1484-91; L11, 1060-62; L13, 547-50; L14, 1993-2002; L17, 1260-95; L18, 2307-20; L19, 1741-58; L21, 544-48*; 1198-1248; L23, 63-65, 137-41*; L24, 99-107*; L27, 2272-83, 2320-23*; 2332-37*; L28, 195-242, 368-424; L29, 101-02*; L30, 931-38; L32, 2878-90; L33, 2422-24. Cross-links to 5S RNA were observed with L5 (positions 34-41), and L18 (precise site not localised).
Osswald
M
M
Max-Planck-Institut für Molekulare Genetik, Abteilung Wittmann, Berlin-Dahlem, FRG.
Greuer
B
B
Brimacombe
R
R
D001483:Base Sequence; D003432:Cross-Linking Reagents; D015180:Electrophoresis, Gel, Two-Dimensional; D004926:Escherichia coli / Q000235:genetics*; D007096:Imidoesters / Q000494:pharmacology*; D008466:Mechlorethamine / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012329:RNA, Bacterial / Q000737:chemistry / Q000235:genetics; D012338:RNA, Ribosomal, 23S / Q000737:chemistry* / Q000235:genetics; D012341:RNA, Ribosomal, 5S / Q000737:chemistry* / Q000235:genetics; D012269:Ribosomal Proteins / Q000737:chemistry*
D003432:Cross-Linking Reagents; D007096:Imidoesters; D012329:RNA, Bacterial; D012338:RNA, Ribosomal, 23S; D012341:RNA, Ribosomal, 5S; D012269:Ribosomal Proteins; D008466:Mechlorethamine; C005905:methyl 4-mercaptobutyrimidate; C028066:methyl 4-azidophenylacetimidate