Open AccessA carbon-13 neclear magnetic resonance study of the 3'-terminus of 16S ribosomal RNA ofEscherichia colispecifically labeled with carbon-13 in the methlgroups of the m26Am26A sequence
Author(s) -
Rein Van Charldorp,
Jennifer Verhoeven,
P.H. Van Knippenberg,
C. A. G. Haasnoot,
Cornelis W. Hilbers
Publication year - 1982
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/10.14.4237
Subject(s) - biology , ribosome , rna , ribosomal rna , escherichia coli , 50s , microbiology and biotechnology , 30s , biochemistry , 16s ribosomal rna , stereochemistry , gene , chemistry
30S ribosomes were isolated from a kasugamycin resistant mutant of E. coli that lacks methylgroups on two adjacent adenines in 16S ribosomal RNA. These ribosomes were methylated in vitro with a purified methylating enzyme and 5-S-adenosyl-(13C-methyl)-L-methionine chloride ((13C-methyl)-SAM) as methyldonor. After in situ cleavage of the 16S ribosomal RNA by the bacteriocin cloacin DF13, the 49 nucleotide fragment from the 3'-end of the RNA was isolated. The carbon-13 nuclear magnetic resonance spectra of the fragment at various temperatures were compared with those of 6-N-dimethyladenosine (m6(2)A) and 6-N-dimethyladenylyl-(3' leads to 5')-6-N-dimethyladenosine (m6(2)Am6(2)A). The data show that the two methylated adenines, which are part of a four membered hairpin loop, show a strong tendency to be stacked in analogy to the dinucleotide m6(2)Am6(2).