23S rRNA domain V, a fragment that can be specifically methylated in vitro by the ErmSF (TlrA) methyltransferase.
The DNA sequence that encodes 23S rRNA domain V of Bacillus subtilis, nucleotides 2036 to 2672 (C. J. Green, G. C. Stewart, M. A. Hollis, B. S. Vold, and K. F. Bott, Gene 37:261-266, 1985), was cloned and used as a template from which to transcribe defined domain V RNA in vitro. The RNA transcripts served as a substrate in vitro for specific methylation of B. subtilis adenine 2085 (adenine 2058 in Escherichia coli 23S rRNA) by the ErmSF methyltransferase, an enzyme that confers resistance to the macrolide-lincosamide-streptogramin B group of antibiotics on Streptomyces fradiae NRRL 2702, the host from which it was cloned. Thus, neither RNA sequences belonging to domains other than V nor the association of 23S rRNA with ribosomal proteins is needed for the specific methylation of adenine that confers resistance to the macrolide-lincosamide-streptogramin B group of antibiotics.
Department of Pharmacology, University of Wisconsin Medical School, Madison 53706.
D
D
Kovalic
R B
RB
Giannattasio
H J
HJ
Jin
B
B
Weisblum
D000225:Adenine / Q000031:analogs & derivatives / Q000302:isolation & purification; D001412:Bacillus subtilis / Q000235:genetics; D001419:Bacteria / Q000201:enzymology* / Q000235:genetics*; D001483:Base Sequence; D002474:Cell-Free System; D017931:DNA Primers; D004926:Escherichia coli / Q000235:genetics; D008745:Methylation; D008780:Methyltransferases / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D012338:RNA, Ribosomal, 23S / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D017423:Sequence Analysis, RNA; D013302:Streptomyces / Q000201:enzymology; D013379:Substrate Specificity
D000225:Adenine; D017931:DNA Primers; D008780:Methyltransferases; D012338:RNA, Ribosomal, 23S; D011994:Recombinant Proteins; C001020:N(6),N(6)-dimethyladenine; C035181:rRNA (adenosine-O-2'-)methyltransferase; C000730567:6-methyladenine