Mutations in the peptidyl transferase center of 23 S rRNA reveal the site of action of sparsomycin, a universal inhibitor of translation.
Sparsomycin is a universal and powerful inhibitor of peptide bond formation which, in contrast to many other ribosome-targeted antibiotics, does not produce footprints on rRNA. A mutant of an archaeon Halobacterium halobium has been isolated that exhibits resistance to sparsomycin. Resistant cells possessed a mutation in the 23 S rRNA, where C2518 (C2499 in Escherichia coli) was substituted by U. Introduction of the C2518U mutation into the chromosomal 23 S rRNA gene of wild-type H. halobium rendered cells resistant to sparsomycin, demonstrating that a single nucleotide alteration in the rRNA is sufficient to confer resistance. Accordingly, ribosomes containing mutant 23 S rRNA exhibited increased tolerance to sparsomycin in vitro. Mutations of two other nucleotide positions in the peptidyl transferase center, C2471 and U2519 (C2452 and U2500 in E. coli), conferred resistance to low concentrations of sparsomycin. The location of the sparsomycin resistance mutations reveals the possible site of drug binding and/or action. Our findings provide further support for the idea that rRNA may be directly involved in interaction with antibiotics and the catalysis of the peptide bond formation.
Tan
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Center for Pharmaceutical Biotechnology-m/c870, University of Illinois, Chicago 60607-7173, USA.
DeBlasio
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Mankin
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D001483:Base Sequence; D001665:Binding Sites; D004252:DNA Mutational Analysis; D004352:Drug Resistance, Microbial / Q000235:genetics; D019617:Halobacterium salinarum / Q000187:drug effects* / Q000235:genetics; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010441:Peptide Chain Elongation, Translational / Q000187:drug effects*; D010458:Peptidyl Transferases / Q000378:metabolism; D017354:Point Mutation; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology*; D012338:RNA, Ribosomal, 23S / Q000737:chemistry* / Q000235:genetics; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism; D012270:Ribosomes / Q000737:chemistry / Q000187:drug effects; D013033:Sparsomycin / Q000494:pharmacology*
D011500:Protein Synthesis Inhibitors; D012338:RNA, Ribosomal, 23S; D012346:RNA, Transfer, Amino Acyl; D013033:Sparsomycin; D010458:Peptidyl Transferases