Mutations in 23S rRNA and Ribosomal Protein L4 Account for Resistance in Pneumococcal Strains Selected In Vitro by Macrolide Passage

Tait-Kamradt, Amelia; Davies, Todd A.; Cronan, Melissa; Jacobs, Michael; Appelbaum, Peter C.; Sutcliffe, Joyce A.

doi:10.1128/aac.44.8.2118-2125.2000

Cited by 261 publications

(269 citation statements)

References 54 publications

(55 reference statements)

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“…In general, mechanisms underlying resistance against these antimicrobials are known to involve active efflux of the antibiotics and modification of the ribosomal target by enzymes such as rRNA methylase, enzymic inactivation or mutations in the 23S rRNA gene, as described previously in various organisms by Chisholm et al (2010), Li et al (2011) and Versalovic et al (1996). Another mechanism, involving point mutations in genes encoding the ribosomal proteins L4 (rplD) and L22 (rplV), has been reported in some clinical isolates (Matsuoka et al, 2004;Tait-Kamradt et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

Molecular mechanism of macrolide–lincosamide resistance in Moraxella catarrhalis

Saito

Nonaka

Nishiyama

et al. 2012

Journal of Medical Microbiology

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We identified a Moraxella catarrhalis strain with high-level resistance to azithromycin (MIC.256 mg l "1 ), NSH1, isolated from nasopharyngeal swab samples from an inpatient with acute bronchitis in a Japanese hospital in 2011 and determined its mechanism of macrolidelincosamide resistance. Antimicrobial susceptibility of M. catarrhalis strains was determined using the Etest and agar dilution methods. Mutations in the four 23S rRNA alleles, the ribosomal proteins L4 and L22, and methylase genes erm(B) and erm(F) were tested by PCR and/or sequencing. The efflux system was examined using appropriate inhibitors. Transformation experiments were performed using DNA amplicons of the 23S rRNA gene of M. catarrhalis strain NSH1. This strain showed high-level resistance to erythromycin, clarithromycin, azithromycin, clindamycin (MICs.256 mg l "1 ) and josamycin (MIC5128 mg l "1 ), and contained the A2058T mutation (Escherichia coli numbering) in four of the 23S rRNA alleles. Mutation of the ribosomal proteins and overproduction of the efflux system were not observed, and methylase genes were not detected. When amplified DNA containing the single A2058T mutation was transformed into M. catarrhalis strains, transformants with three A2058T-mutated 23S rRNA alleles showed highlevel resistance to macrolide-lincosamide, similar to strain NSH1. In contrast, transformants with two A2058T-mutated 23S rRNA alleles showed low-level MICs (azithromycin: 0.38-0.5 mg l "1 ). Thus, a single A2058T mutation occurring in at least three 23S rRNA alleles confers high-level resistance to 14-, 15-and 16-membered macrolides and lincosamides in M. catarrhalis possessing four 23S rRNA alleles. This study represents the first evidence, to our knowledge, of this effect in M. catarrhalis.

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Section: Introductionmentioning

confidence: 99%

Molecular mechanism of macrolide–lincosamide resistance in Moraxella catarrhalis

Saito

Nonaka

Nishiyama

et al. 2012

Journal of Medical Microbiology

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“…Macrolide resistance in Streptococcus pneumoniae is due to either target site modification-generally depending on a posttranscriptional modification of 23S rRNA mediated by ermclass methylases (39) or less often on mutations in 23S rRNA or ribosomal proteins (7,37,38)-or active efflux. In this study, 120 erythromycin-resistant pneumococci were investigated for macrolide resistance phenotypes and genotypes, with emphasis on the discrimination between mef(A) and mef(E).…”

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confidence: 99%

Phenotypes and Genotypes of Erythromycin-Resistant Pneumococci in Italy

et al. 2003

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Of 120 erythromycin-resistant pneumococci isolated in Italian hospitals, 39 (32.5%) were M-type isolates, carrying the mef gene alone. The mef gene was also detected, together with erm(AM), in one constitutively resistant isolate and in five isolates of the partially inducible phenotype. Among the 45 mef-positive isolates, 25 (55.6%) carried mef(A) and 20 (44.4%) carried mef(E) as observed from PCR-restriction fragment length polymorphism analysis of a 1,743-bp amplicon. The same result was obtained by a similar method applied to a more common 348-bp amplicon.Macrolide resistance in Streptococcus pneumoniae is due to either target site modification-generally depending on a posttranscriptional modification of 23S rRNA mediated by ermclass methylases (39) or less often on mutations in 23S rRNA or ribosomal proteins (7, 37, 38)-or active efflux. In this study, 120 erythromycin-resistant pneumococci were investigated for macrolide resistance phenotypes and genotypes, with emphasis on the discrimination between mef(A) and mef(E).Efflux-mediated resistance. The efflux mechanism, which reduces the intracellular antibiotic concentration to subtoxic levels (40), is associated with a resistance pattern (M phenotype) characterized by resistance, among macrolides-lincosamidesstreptogramin B (MLS), only to 14-and 15-membered macrolides, usually at a low level (34). M-type resistance is mediated by the mef gene, two variants of which are conventionally described: one, mef(A), originally discovered in Streptococcus pyogenes (5), and the other, mef(E), originally discovered in S. pneumoniae (36). Considering that the mef genes are detected mostly by a PCR method unable to distinguish between the two variants (33) and that mef(A) and mef(E) have 90% identity (36), they are regarded as a single gene class, designated mef(A) (28). However, mef(A) and mef(E) have recently been shown to be carried by different genetic elements in S. pneumoniae, which are inserted at different sites in the chromosome. Both elements carry, adjacent to mef, an open reading frame showing homology to the msr(A) gene associated with macrolide efflux in Staphylococcus aureus (6,12,29). Due to a number of important differences in the properties of mef(A)-and mef(E)-carrying pneumococci, it has been recommended that the distinction between the two genes be maintained (6).Bacterial strains and typing. The 120 erythromycin-resistant pneumococci studied (for which the MIC of erythromycin was Ն1 g/ml) were independent isolates, collected from several laboratories throughout Italy between 1999 and 2002. All were clinical strains, isolated from a variety of clinical specimens (upper respiratory tract material, sputum, blood, cerebrospinal fluid). Multiple isolates from the same patient were excluded.Strain identification was confirmed in our laboratory by conventional tests such as susceptibility to optochin and solubility in bile and by employing the API system (BioMérieux, Marcyl'Etoile, France). Serotyping, done by the capsular swelling test using specific anti...

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“…It is noteworthy that, in mutans streptococci, mutations in the 23S rRNA and rplC genes were first identified in S. criceti strain OMZ 61. In S. pneumoniae, a single A2058G mutation was sufficient to cause elevation of erythromycin MICs, but macrolide resistance was conferred only when two or more alleles harbored an A2058G mutation (Tait-Kamradt et al, 2000). Direct sequencing analysis and sequence analysis of clones indicated that the A2059G mutation (A2058G in E. coli numbering) is predominant in alleles of strain OMZ 61.…”

Section: Discussionmentioning

confidence: 99%

“…DNA hybridization analysis of the 23S rRNA genes in S. criceti strains As mutations of the 23S rRNA genes and the rplD gene of S. pneumoniae are associated with resistance to macrolides (Tait-Kamradt et al, 2000;Wierzbowski et al, 2007), we first examined the 23S rRNA genes (rrl genes) of the four S. criceti strains by DNA hybridization. Genome sequence analysis of strain HS-6 T indicated that the genome contains five rrl genes and that five fragments, 12.8, 9.4, 9.2, 7.9 and 7.1 kb in size, should be detected after EcoRI digestion, whereas five fragments, 7.4, 6.0, 5.3, 3.8 and 3.3 kb in size, should be detected after EcoRI and BbsI double digestion (Richards Fig.…”

Section: Susceptibility Of S Criceti Strains To Antibioticsmentioning

confidence: 99%

“…DOI: http://doi.org/10.1266/ggs.15-00038 (Takaya et al, 2013). Furthermore, mutations in the 23S rRNA genes and the rplD gene encoding the ribosomal protein L4 have been found in clinical isolates and laboratory-generated mutants of macrolide-resistant S. pneumoniae (Tait-Kamradt et al, 2000;Wierzbowski et al, 2007). Of note is that mutation of A2058 in the 23S rRNA gene confers resistance to macrolide, lincosamide and streptogramin type B in S. pneumoniae as well as in E. coli, Helicobacter pylori, Propionibacteria and Streptomyces ambofaciens (for review, see Vester and Douthwaite, 2001).…”

Section: Introductionmentioning

confidence: 99%

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Identification of A2059G 23S rRNA and G439A <i>rplC</i> gene mutations in <i>Streptococcus criceti</i> strain OMZ 61, a strain resistant to azithromycin, josamycin and clindamycin

2015

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Streptococcus criceti is a cariogenic organism that belongs to the mutans streptococci. Of the four S. criceti strains, strain OMZ 61 has been identified as being resistant to erythromycin. Antimicrobial susceptibility testing showed that strain OMZ 61 is also resistant to azithromycin, josamycin and clindamycin but susceptible to tetracycline and tiamulin. DNA hybridization analysis of the 23S rRNA genes revealed that the hybridization patterns in strain OMZ 61 differed from those in the other three strains. We further analyzed the nucleotide sequences of a ribosomal RNA operon, the rrnD operon, and the rpsJ−rpsQ region including rplC and rplD genes for ribosomal proteins L3 and L4, respectively, in the four strains studied. Nucleotide sequence analysis indicated that strain OMZ 61 contains an A-to-G substitution at nucleotide position 2059, equivalent to Escherichia coli numbering 2058, in a 23S rRNA gene (rrlD) and a G-to-A substitution at nucleotide position 439 in the rplC gene, suggesting an amino acid residue change at position 147 from valine to isoleucine, whereas no mutation in the rplD gene was found. DNA sequencing and polymerase chain reaction-restriction fragment length polymorphism analysis showed that most or all of the 23S rRNA genes in strain OMZ 61 contain the A2059G mutation. These findings suggest that the resistance to erythromycin, azithromycin, josamycin and clindamycin in strain OMZ 61 is conferred by alterations in 23S rRNA and/or ribosomal protein L3. This is the first description of mutations in the 23S rRNA and rplC genes in mutans streptococci.

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Mutations in 23S rRNA and Ribosomal Protein L4 Account for Resistance in Pneumococcal Strains Selected In Vitro by Macrolide Passage

Cited by 261 publications

References 54 publications

Molecular mechanism of macrolide–lincosamide resistance in Moraxella catarrhalis

Molecular mechanism of macrolide–lincosamide resistance in Moraxella catarrhalis

Phenotypes and Genotypes of Erythromycin-Resistant Pneumococci in Italy

Identification of A2059G 23S rRNA and G439A <i>rplC</i> gene mutations in <i>Streptococcus criceti</i> strain OMZ 61, a strain resistant to azithromycin, josamycin and clindamycin

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