The three-dimensional folding of ribosomal RNA; localization of a series of intra-RNA cross-links in 23S RNA induced by treatment of Escherichia coli 50S ribosomal subunits with bis-(2-chloroethyl)-methylamine.
Intact 50S ribosomal subunits from E.coli were cross-linked with the symmetrical bifunctional reagent bis-(2-chloroethyl)-methylamine. After deproteinization, selected regions of the 23S RNA were excised by treatment with ribonuclease H in the presence of appropriate complementary decadeoxynucleotides, and screened for the presence of intra-RNA cross-links by two-dimensional gel electrophoresis. Individual isolated cross-linked RNA fragments were analysed by our established procedures. Sixteen intra-RNA cross-links were identified, three of which corresponded to those previously published. The thirteen 'new' cross-links were localized in the 23S RNA at positions 774-78 linked to 792-94, 876-79 linked to 899-900, 979-81 or 983-84 to 2029, 1715 to 1743-46, 1911-21 to 1964, 1933 to 1966, 2032 to 2054-55, 2112 to 2169-71, 2116-17 to 2163-67, 2128-32 to 2156-59, 2392-93 to 2422-23, 2737-38 to 2763-66, and 2791 to 2890. These results are discussed in the context of three-dimensional model-building studies with the 23S RNA, with particular reference to the environment of the 'active centre' of the 50S subunit.
Döring
T
T
Max-Planck-Institut für Molekulare Genetik, Abteilung Wittmann, Berlin, FRG.
Greuer
B
B
Brimacombe
R
R
D001483:Base Sequence; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D015180:Electrophoresis, Gel, Two-Dimensional; D004926:Escherichia coli; D008466:Mechlorethamine / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation / Q000187:drug effects*; D012329:RNA, Bacterial / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D012338:RNA, Ribosomal, 23S / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D012260:Ribonucleases / Q000378:metabolism; D012270:Ribosomes / Q000187:drug effects*
D003432:Cross-Linking Reagents; D012329:RNA, Bacterial; D012338:RNA, Ribosomal, 23S; D008466:Mechlorethamine; D012260:Ribonucleases