Mutagenesis of 16S rRNA C1409-G1491 Base-pair Differentiates Between 6′OH and 6′NH3+ Aminoglycosides

Pfister, Peter; Hobbie, Sven N.; Brüll, Christian; Corti, Natascia; Vasella, Andrea; Westhof, Éric; Böttger, Erik C.

doi:10.1016/j.jmb.2004.11.073

Cited by 63 publications

(83 citation statements)

References 28 publications

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“…Strains with mutations in 16S rRNA residues 1408, 1409 and 1491 show a mutation-specific drug susceptibility pattern that is independent of the amino acid residue 42 in ribosomal protein S12 (wt vs K42R, see Table 1) and that corresponded to previously published data [12]. These results demonstrate that the K42R mutation in ribosomal protein S12 does not affect the susceptibility of M. smegmatis H44 mutants to 2-deoxystreptamines.…”

Section: Resultssupporting

confidence: 85%

“…Mutagenesis of the H44 decoding-site RNA was performed in strain M. smegmatis Δ rrnB (SZ380) and in strain M. smegmatis Δ rrnB rpsL K42R (SZ004) [12]. rRNA mutations were generated by PCR, cloned into vector pMIH-rrnB and introduced into the 16S rRNA A-site of the single rRNA allelic strain SZ0380 by RecA-mediated homologous recombination as described [12].…”

Section: Methodsmentioning

confidence: 99%

“…rRNA mutations were generated by PCR, cloned into vector pMIH-rrnB and introduced into the 16S rRNA A-site of the single rRNA allelic strain SZ0380 by RecA-mediated homologous recombination as described [12]. For a list of strains and plasmids see Tables S1 and S2 in Supplementary Data.…”

Section: Methodsmentioning

confidence: 99%

“…MIC tests were performed in a microtiter plate format as described [12]. In brief, freshly grown M. smegmatis cultures were resuspended in LB broth supplemented with 0.05% of Tween 80, diluted to an absorbance at 600 nm of 0.025 and incubated in the presence of 2-fold serial dilutions of 2-deoxystreptamine aminoglycosides.…”

Section: Methodsmentioning

confidence: 99%

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Mutation K42R in Ribosomal Protein S12 Does Not Affect Susceptibility of Mycobacterium smegmatis 16S rRNA A-Site Mutants to 2-Deoxystreptamines

et al. 2010

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Recent studies have suggested that ribosomal protein S12 modulates 16S rRNA function and susceptibility to 2-deoxystreptamine aminoglycosides. To study whether the non-restrictive K42R mutation in RpsL affects 2-deoxystreptamine susceptibility in Mycobacterium smegmatis, we studied the drug susceptibility pattern of various mutants with genetic alterations in the 16S rRNA decoding A-site in the context of wild-type and mutant protein S12. RpsL K42R substitution was found not to affect the drug resistance pattern associated with mutational alterations in 16S rRNA H44.

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Section: Resultssupporting

confidence: 85%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Mutation K42R in Ribosomal Protein S12 Does Not Affect Susceptibility of Mycobacterium smegmatis 16S rRNA A-Site Mutants to 2-Deoxystreptamines

et al. 2010

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“…However, the A1408G mutation in E. coli 16S rRNA conferred only an intermediate level of resistance to aminoglycosides with a hydroxyl group at the 69 position of ring I, such as paromomycin and geneticin (G418) (Recht et al 1999b;Gregory et al 2005). In contrast, mutations at G1491 conferred much greater resistance to these compounds (Pfister et al 2005b). These results suggested that G1491 also plays an important role in determining bacterial sensitivity to aminoglycosides.…”

Section: Introductionmentioning

confidence: 99%

Eukaryotic ribosomal RNA determinants of aminoglycoside resistance and their role in translational fidelity

Fan-Minogue¹,

Bedwell²

2007

RNA

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Recent studies of prokaryotic ribosomes have dramatically increased our knowledge of ribosomal RNA (rRNA) structure, functional centers, and their interactions with antibiotics. However, much less is known about how rRNA function differs between prokaryotic and eukaryotic ribosomes. The core decoding sites are identical in yeast and human 18S rRNAs, suggesting that insights obtained in studies with yeast rRNA mutants can provide information about ribosome function in both species. In this study, we examined the importance of key nucleotides of the 18S rRNA decoding site on ribosome function and aminoglycoside susceptibility in Saccharomyces cerevisiae cells expressing homogeneous populations of mutant ribosomes. We found that residues G577, A1755, and A1756 (corresponding to Escherichia coli residues G530, A1492, and A1493, respectively) are essential for cell viability. We also found that residue G1645 (A1408 in E. coli ) and A1754 (G1491 in E. coli ) both make significant and distinct contributions to aminoglycoside resistance. Furthermore, we found that mutations at these residues do not alter the basal level of translational accuracy, but influence both paromomycin-induced misreading of sense codons and readthrough of stop codons. This study represents the most comprehensive mutational analysis of the eukaryotic decoding site to date, and suggests that many fundamental features of decoding site function are conserved between prokaryotes and eukaryotes.

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Molecular Recognition of RNA by Neomycin and a Restricted Neomycin Derivative

Zhao¹,

Zhao²,

Blount³

et al. 2005

Angewandte Chemie

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show abstract

Mutagenesis of 16S rRNA C1409-G1491 Base-pair Differentiates Between 6′OH and 6′NH3+ Aminoglycosides

Cited by 63 publications

References 28 publications

Mutation K42R in Ribosomal Protein S12 Does Not Affect Susceptibility of Mycobacterium smegmatis 16S rRNA A-Site Mutants to 2-Deoxystreptamines

Mutation K42R in Ribosomal Protein S12 Does Not Affect Susceptibility of Mycobacterium smegmatis 16S rRNA A-Site Mutants to 2-Deoxystreptamines

Eukaryotic ribosomal RNA determinants of aminoglycoside resistance and their role in translational fidelity

Molecular Recognition of RNA by Neomycin and a Restricted Neomycin Derivative

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