Streptomycin binds to the decoding center of 16 S ribosomal RNA.
Streptomycin, an error-inducing aminoglycoside antibiotic, binds to a single site on the small ribosomal subunit of bacteria, but this site has not yet been defined precisely. Here, we demonstrate that streptomycin binds to E. coli 16 S rRNA in the absence of ribosomal proteins, and protects a set of bases in the decoding region against dimethyl sulfate attack. The binding studies were performed in a high ionic strength buffer containing 20 mM Mg2+. The pattern of protection in the decoding region was similar to that observed when streptomycin binds to the 30 S subunit. However, streptomycin also protects the 915 region of 16 S rRNA within the 30 S subunit, whereas it did not protect the 915 region of the naked 16 S rRNA. The interaction of streptomycin with 16 S rRNA was further defined by using two fragments that correspond to the 3' minor domain of 16 S rRNA and to the decoding analog, a portion of this domain encompassing the decoding center. In the presence of streptomycin, the pattern of protection against dimethyl sulfate attack for the two fragments was similar to that seen with the full-length 16 S rRNA. This indicates that the 3' minor domain as well as the decoding analog contain the recognition signals for the binding of streptomycin. However, streptomycin could not bind to the decoding analog in the absence of Mg2+. This contrasts with neomycin, another error-inducing aminoglycoside antibiotic, that binds to the decoding analog in the absence of Mg2+, but not at 20 mM Mg2+. Our results suggest that both neomycin and streptomycin interact with the decoding center, but recognize alternative conformations of this region.
Spickler
C
C
Département de Biochimie, Université de Montréal, Montréal, Québec, H3C 3J7, Canada.
Brunelle
M N
MN
Brakier-Gingras
L
L
D000900:Anti-Bacterial Agents / Q000378:metabolism* / Q000494:pharmacology; D001483:Base Sequence; D001665:Binding Sites; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000378:metabolism; D008274:Magnesium / Q000494:pharmacology; D008969:Molecular Sequence Data; D009153:Mutagens / Q000494:pharmacology; D009355:Neomycin / Q000378:metabolism / Q000494:pharmacology; D009690:Nucleic Acid Conformation; D012329:RNA, Bacterial / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D012336:RNA, Ribosomal, 16S / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D013307:Streptomycin / Q000378:metabolism* / Q000494:pharmacology; D013463:Sulfuric Acid Esters / Q000494:pharmacology
D000900:Anti-Bacterial Agents; D009153:Mutagens; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D013463:Sulfuric Acid Esters; D009355:Neomycin; D008274:Magnesium; C007482:dimethyl sulfate; D013307:Streptomycin