Mutation of L4 Ribosomal Protein Conferring Unusual Macrolide Resistance in Two Independent Clinical Isolates of<i>Staphylococcus aureus</i>

Prunier, Anne-Laure; Trong, Hiep N′Guyen; Tandé, Didier; Segond, Christine; Leclercq, Roland

doi:10.1089/mdr.2005.11.18

Cited by 22 publications

(21 citation statements)

References 13 publications

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“…Our isolation of a Lys68Gln mutant is the first report of an L4 mutation in S. aureus associated with resistance through selection with oxazolidinones. Further studies are needed to conclusively link this mutation to the observed increase in LZD resistance; however, a number of other documented L4 mutations involve alterations in amino acids in this region of the protein (18,23,56,76). Mutations in the rplC gene encoding ribosomal protein L3 were found in a large portion of the mutants selected in our spontaneous mutation frequency experiments.…”

Section: ϫ10mentioning

confidence: 93%

Novel Ribosomal Mutations in Staphylococcus aureus Strains Identified through Selection with the Oxazolidinones Linezolid and Torezolid (TR-700)

Locke

Hilgers

Shaw

2009

Antimicrob Agents Chemother

182

146

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TR-700 (torezolid), the active moiety of the novel oxazolidinone phosphate prodrug TR-701, is highly potent against gram-positive pathogens, including strains resistant to linezolid (LZD). Here we investigated the potential of Staphylococcus aureus strains ATCC 29213 (methicillin-susceptible S. aureus [MSSA]) and ATCC 33591 (methicillin-resistant S. aureus [MRSA]) to develop resistance to TR-700. The spontaneous frequencies of mutation of MSSA 29213 and MRSA 33591 resulting in reduced susceptibility to TR-700 at 2؋ the MIC were 1.1 ؋ 10 ؊10 and 1.9 ؋ 10 ؊10 , respectively. These values are ϳ16-fold lower than the corresponding LZD spontaneous mutation frequencies of both strains. Following 30 serial passages in the presence of TR-700, the MIC for MSSA 29213 remained constant (0.5 g/ml) while increasing eightfold (0.25 to 2.0 g/ml) for MRSA 33591. Serial passage of MSSA 29213 and MRSA 33591 in LZD resulted in 64-and 32-fold increases in LZD resistance (2 to 128 g/ml and 1 to 32 g/ml, respectively). Domain V 23S rRNA gene mutations (Escherichia coli numbering) found in TR-700-selected mutants included T2500A and a novel coupled T2571C/G2576T mutation, while LZD-selected mutants included G2447T, T2500A, and G2576T. We also identified mutations correlating with decreased susceptibility to TR-700 and LZD in the rplC and rplD genes, encoding the 50S ribosomal proteins L3 and L4, respectively. L3 mutations included Gly152Asp, Gly155Arg, Gly155Arg/ Met169Leu, and ⌬Phe127-His146. The only L4 mutation detected was Lys68Gln. TR-700 maintained a fourfold or greater potency advantage over LZD against all strains with ribosomal mutations. These data bring to light a variety of novel and less-characterized mutations associated with S. aureus resistance to oxazolidinones and demonstrate the low resistance potential of torezolid.Staphylococcus aureus infections pose a serious health threat worldwide. Increasing antibiotic resistance and the prevalence of methicillin (meticillin)-resistant S. aureus (MRSA) in clinical settings have created a demand for novel therapeutic agents. Linezolid (LZD) has a broad spectrum of activity against a variety of gram-positive pathogens, including MRSA, and was the first oxazolidinone antibiotic to gain FDA approval (1). LZD acts through inhibition of protein synthesis via binding to the peptidyl transferase center (PTC) of the 50S ribosomal subunit (37,65,68). Despite in vitro studies demonstrating a low resistance potential for LZD (31, 79), soon after its approval in 2000, LZD-resistant (LZD r ) MRSA (72) and LZD r , vancomycin (VAN)-resistant enterococci (22) emerged in the clinic. Although rare, resistance has most commonly occurred in patients undergoing long-term LZD therapy (10,17,22,45,72,74). Three classes of oxazolidinone resistance mechanisms have been previously characterized: mutations in the domain V region of 23S rRNA genes (69), acquisition of the ribosomal methyltransferase gene cfr (43), and mutations in the rplD gene encoding the 50S ribosomal protein L4 (76).A variety of 23S ...

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Section: ϫ10mentioning

confidence: 93%

Novel Ribosomal Mutations in Staphylococcus aureus Strains Identified through Selection with the Oxazolidinones Linezolid and Torezolid (TR-700)

Locke

Hilgers

Shaw

2009

Antimicrob Agents Chemother

182

146

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“…In H. influenzae, L4 amino acid variations (insertion 65 GT, K 61 Q, T 64 K, G 65 D; deletion 65 GR, G 53 A; deletion of 66 RA, A 69 S, T 82 I, D 94 E, D 139 G), some outside the loop region, could provide high-level resistance of up to 128 mg/mL for 14-and 15-membered macrolides (Clark et al 2002;Peric et al 2003). S. aureus isolates with L4 amino acid changes R 168 S, G 69 A, and T 70 P have been described in CF patients (Prunier et al 2005). Amino acid variations in L4 from M. genitalium (N 21 K, H 69 R, V 84 G, E 128 G, P 81 S, Y 135 P, N 172 S, N 172 S, A 114 V, A 116 V, A 114 S, R 45 K) were found encoded in DNA from the urine of men with nongonococcal urethritis , and in the chromosomal DNA isolated from a collection of M. genitalium isolates .…”

Section: Ribosomal Protein Mutationsmentioning

confidence: 99%

Resistance to Macrolide Antibiotics in Public Health Pathogens

Fyfe

Grossman

Kerstein

et al. 2016

Cold Spring Harb Perspect Med

163

140

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Macrolide resistance mechanisms can be target-based with a change in a 23S ribosomal RNA (rRNA) residue or a mutation in ribosomal protein L4 or L22 affecting the ribosome's interaction with the antibiotic. Alternatively, mono-or dimethylation of A2058 in domain V of the 23S rRNA by an acquired rRNA methyltransferase, the product of an erm (erythromycin ribosome methylation) gene, can interfere with antibiotic binding. Acquired genes encoding efflux pumps, most predominantly mef(A) þ msr(D) in pneumococci/streptococci and msr(A/B) in staphylococci, also mediate resistance. Drug-inactivating mechanisms include phosphorylation of the 2 0 -hydroxyl of the amino sugar found at position C5 by phosphotransferases and hydrolysis of the macrocyclic lactone by esterases. These acquired genes are regulated by either translation or transcription attenuation, largely because cells are less fit when these genes, especially the rRNA methyltransferases, are highly induced or constitutively expressed. The induction of gene expression is cleverly tied to the mechanism of action of macrolides, relying on antibiotic-bound ribosomes stalled at specific sequences of nascent polypeptides to promote transcription or translation of downstream sequences. Macrolide antibiotics are polyketides composed of a 14-, 15-, or 16-membered macrocyclic lactone ring (14-, 15-, and 16-membered) to which several sugars and/or side chains have been attached by the producing organism or as modifications during semisynthesis in the laboratory (Figs. 1 and 2). Newer semisynthetic derivations, like ketolides telithromycin, and solithromycin, have a C3-keto group in place of the C3 cladinose (akin to naturally occurring pikromycin) (Brockmann and Henkel 1950) and an 11,12-cyclic carbamate with an extended alkyl -aryl side chain that increases the affinity of the antibiotic for the ribosome by 10-to 100-fold (Hansen et al. 1999;Dunkle et al. 2010); in the case of solithromycin, a fluorine substituent at C2 provides an additional ribosomal interaction (Llano-Sotelo et al. 2010). Macrolides continue to be important in the therapeutic treatment of community-acquired pneumonia (Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, and atypicals Legionella pneumophila, Mycoplasma pneumoniae, Chlamydia pneumoniae), sexually transmitted diseases (Neiserria gonorhoeae, Chlamydia trachomatis, Mycoplasma genitalium), shigellosis, and salmonellosis. With solithromycin heading for a new drug application (NDA) filing in 2016 and having the in vitro potency to treat erythromycin-resistant

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“…Ribosomal alteration is mediated by a ) and in ribosomal proteins L4 and L22 (encoded by rplD and rplV genes, respectively) have been infrequently described [11][12][13]. Active efflux is mediated by the msr(A) gene that codes for a putative efflux pump and is responsible for the so-called M phenotype characterised by unique resistance to 14-and 15-membered ring macrolides.…”

Section: Introductionmentioning

confidence: 99%

Molecular basis of resistance to macrolides, lincosamides and streptogramins in Staphylococcus saprophyticus clinical isolates

Bouter

Leclercq

Cattoir

2011

International Journal of Antimicrobial Agents

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The aim of this study was to evaluate the prevalence of resistance to macrolide-lincosamide-streptogramin (MLS) antibiotics as well as to assess the molecular basis of this resistance amongst 72 Staphylococcus saprophyticus urinary isolates collected from 2005 to 2009 in University Hospital of Caen (France). Of the 72 strains studied, 33 (45.8%) were resistant to at least one MLS antibiotic, including 24 (72.7%) with an M phenotype, 5 (15.2%) with an inducible MLS(B) phenotype, 3 (9.1%) with a combined M+L phenotype and 1 (3.0%) with an L phenotype. All isolates were susceptible to the combination of streptogramins A and B. The resistance genes erm(A), erm(B), erm(C), msr(A) and lnu(A) were detected alone in 0, 0, 5 (15.2%), 24 (72.7%) and 1 (3.0%) of the 33 MLS-resistant isolates, respectively, whereas 2 strains (6.1%) were positive for both msr(A) and lnu(A). All msr(A)-positive isolates exhibited an M phenotype, whereas all five erm(C)-positive and all three lnu(A)-positive strains displayed, respectively, an inducible MLS(B) phenotype and an L phenotype with a positive Hodge test. Plasmid analysis indicated that erm(C) and lnu(A) genes were borne by small-size plasmids (ca. 2.5 kb), whereas larger plasmids (30-90 kb) harboured msr(A). In conclusion, these findings show a high prevalence of MLS resistance in S. saprophyticus, which was mainly associated with the presence of the msr(A) gene. Since S. saprophyticus colonises the gastrointestinal tract, it may constitute an unexpected reservoir for MLS resistance genes, in particular msr(A), amongst coagulase-negative staphylococci.

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Mutation of L4 Ribosomal Protein Conferring Unusual Macrolide Resistance in Two Independent Clinical Isolates ofStaphylococcus aureus

Cited by 22 publications

References 13 publications

Novel Ribosomal Mutations in Staphylococcus aureus Strains Identified through Selection with the Oxazolidinones Linezolid and Torezolid (TR-700)

Novel Ribosomal Mutations in Staphylococcus aureus Strains Identified through Selection with the Oxazolidinones Linezolid and Torezolid (TR-700)

Resistance to Macrolide Antibiotics in Public Health Pathogens

Molecular basis of resistance to macrolides, lincosamides and streptogramins in Staphylococcus saprophyticus clinical isolates

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