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Clustering of modified nucleotides at the functional center of bacterial ribosomal RNA
Author(s) -
Brimacombe Richard,
Mitchell Philip,
Osswald Monika,
Stade Katrin,
Bochkariov Dmitry
Publication year - 1993
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.7.1.8422963
Subject(s) - nucleotide , ribosomal rna , cluster analysis , center (category theory) , biology , computational biology , genetics , chemistry , computer science , artificial intelligence , gene , crystallography
An aryl trifluoromethyl diazirine photo‐reactive derivative was attached to the 2‐thiocytidine residue at position 32 of tRNA I Arg and this derivatized tRNA was bound to Escherichia coli 70S ribosomes. After irradiation at 350 nm the site of cross‐linking to the 16S RNA was analyzed by our standard procedures and found to lie within the secondary structural element comprising bases 956‐983; this region contains two modified nucleotides at positions 966 and 967. Similarly, an aryl azido photoreactive derivative was attached to the phenylalanine residue of Phe‐tRNA Phe , and the derivatized aminoacyl tRNA was bound to the ribosome either at the A‐ or the P‐site. In both cases, after irradiation at 250 nm, the cross‐link site was localized to position 2439 of the 23S RNA; in the secondary structure of the latter the neighboring nucleotide 2442 is base‐paired to a modified nucleotide at position 2069. Taken together with other cross‐linking data, these results now directly implicate a total of 27 out of the 29 modified nucleotides in E. coli 16S and 23S RNA as lying within or close to the functional center of the ribosome.— Brimacombe, R., Mitchell, P., Osswald, M., Stade, K., Bochkariov, D. Clustering of modified nucleotides at the functional center of bacterial ribosomal RNA. FASEB J. 7: 161‐167; 1993.