title: Modulation of 16S rRNA function by ribosomal protein S12.
abstract: Ribosomal protein S12 is a critical component of the decoding center of the 30S ribosomal subunit and is involved in both tRNA selection and the response to streptomycin. We have investigated the interplay between S12 and some of the surrounding 16S rRNA residues by examining the phenotypes of double-mutant ribosomes in strains of Escherichia coli carrying deletions in all chromosomal rrn operons and expressing total rRNA from a single plasmid-borne rrn operon. We show that the combination of S12 and otherwise benign mutations at positions C1409-G1491 in 16S rRNA severely compromises cell growth while the level and range of aminoglycoside resistances conferred by the G1491U/C substitutions is markedly increased by a mutant S12 protein. The G1491U/C mutations in addition confer resistance to the unrelated antibiotic, capreomycin. S12 also interacts with the 912 region of 16S rRNA. Genetic selection of suppressors of streptomycin dependence caused by mutations at proline 90 in S12 yielded a C912U substitution in 16S rRNA. The C912U mutation on its own confers resistance to streptomycin and restricts miscoding, properties that distinguish it from a majority of the previously described error-promoting ram mutants that also reverse streptomycin dependence.
authors.affiliation: Berkeley Center for Synthetic Biology, University of California, Berkeley, 717 Potter St., Berkeley, CA 94720-3224, USA.
authors.forename: Anton
authors.initials: A
authors.lastname: Vila-Sanjurjo
authors.forename: Ying
authors.initials: Y
authors.lastname: Lu
authors.forename: Jamie L
authors.initials: JL
authors.lastname: Aragonez
authors.forename: Rebekah E
authors.initials: RE
authors.lastname: Starkweather
authors.forename: Manoj
authors.initials: M
authors.lastname: Sasikumar
authors.forename: Michael
authors.initials: M
authors.lastname: O'Connor
mesh_terms: D000595:Amino Acid Sequence; D002207:Capreomycin / Q000494:pharmacology; D024881:Drug Resistance, Bacterial / Q000235:genetics / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics*; D008958:Models, Molecular; D009154:Mutation; D012336:RNA, Ribosomal, 16S / Q000187:drug effects* / Q000235:genetics / Q000502:physiology*; D012269:Ribosomal Proteins / Q000235:genetics / Q000502:physiology*; D013307:Streptomycin / Q000494:pharmacology
chemical_list: D012336:RNA, Ribosomal, 16S; D012269:Ribosomal Proteins; C034212:ribosomal protein S12; D002207:Capreomycin; D013307:Streptomycin