Nucleic Acids Research, 1988, Vol. 16, No. 20 9631-9639
© 1988
Articles |
A mutation in the 530 loop of Escherichia coli 16S ribosomal RNA causes resistance to streptomycin
Départment de Biochimie, Université de Montréal Montréal, H3C 3J7 1Départment de Biochimie, Université Laval Québec, G1K 7P4, Canada
*To whom correspondence should be addressed
Received June 24, 1988. Accepted September 15, 1988.
Oligonucleotide-directed mutagenesis was used to introduce an A to C transversion at position 523 in the 16S ribosomal RNA gene of Escherichia coli rrnB operon cloned in plasmid pKK3535. E. coli cells transformed with the mutated plasmid were resistant to streptomycin. The mutated ribosomes isolated from these cells were not stimulated by streptomycin to misread the message in a poly(U)-directed assay. They were also restrictive to the stimulation of misreading by other error-promoting related aminoglycoside antibiotics such as neomycin, kanamy-cin or gentamicin, which do not compete for the streptomycin binding site. The 530 loop where the mutation in the 16S rRNA is located has been mapped at the external surface of the 30S subunit, and is therefore distal from the streptomcin binding site at the subunit interface. Our results support the conclusion that the mutation at position 523 in the 16S rRNA does not interfere with the binding of streptomycin, but prevents the drug from inducing conformational changes in the 530 loop which account for its miscoding effect. Since this effect primarily results from a perturbation of the translational proofreading control, our results also provide evidence that the 530 loop of the 16S rRNA is involved in this accuracy control.
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