Localisation of a series of intra–RNA cross-links in the secondary and tertiary structure of 23S RNA, induced by ultraviolet irradiation ofEscherichia50S ribosomal subunits | Zendy

Wolfgang Stiege | Zendy; Carola Gkotz | Zendy; Richard Brimacombe | Zendy

Open Access

Localisation of a series of intra–RNA cross-links in the secondary and tertiary structure of 23S RNA, induced by ultraviolet irradiation ofEscherichia50S ribosomal subunits

Author(s) -

Wolfgang Stiege,

Carola Gkotz,

Richard Brimacombe

Publication year - 1983

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/11.6.1687

Subject(s) - rna , biology , ribosomal rna , oligonucleotide , ribonuclease , microbiology and biotechnology , 50s , 23s ribosomal rna , protein secondary structure , escherichia coli , biochemistry , ribosome , gene

Intra-RNA cross-links were introduced into E. coli 50S ribosomal subunits by mild ultraviolet irradiation. The subunits were partially digested with cobra venom nuclease, and the cross-linked RNA complexes were isolated by two-dimensional electrophoresis. Many of the complexes were submitted to a second partial digestion procedure. Oligonucleotide analysis of the RNA fragments obtained in this manner enabled cross-links between the following ribonuclease T1 oligonucleotides in the 23S RNA to be established: positions 292-296 and 339-350; 601-604 and 652-656; 1018-1022 and 1140-1149; 1433-1435 and 1556-1560; 1836-1839 and 1898-1903; 2832-2834 (tentative) and 2878-2885; 2849-2852 and 2865-2867 (tentative); 739-748 and 2609-2618; 571-577 and 2030-2032; 1777-1792 (tentative) and 2584-2588. The first seven of these cross-links lie within the secondary structure of the 23S RNA, whereas the last three are tertiary structural cross-links. The degree of precision of the individual determinations was variable, depending on the nucleotide sequence in the vicinity of the cross-link site concerned.

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