Vancomycin tolerance in clinical and laboratory Streptococcus pneumoniae isolates depends on reduced enzyme activity of the major LytA autolysin or cooperation between CiaH histidine kinase and capsular polysaccharide.
Vancomycin is frequently added to standard therapy for pneumococcal meningitis. Although vancomycin-resistant Streptococcus pneumoniae strains have not been isolated, reports on the emergence of vancomycin-tolerant pneumococci are a cause of concern. To date, the molecular basis of vancomycin tolerance in S. pneumoniae is essentially unknown. We examined two vancomycin-tolerant clinical isolates, i.e. a purported autolysin negative (LytA(-)), serotype 23F isolate (strain S3) and the serotype 14 strain 'Tupelo', which is considered a paradigm of vancomycin tolerance. S3 was characterized here as carrying a frameshift mutation in the lytA gene encoding the main pneumococcal autolysin. The vancomycin tolerance of strain S3 was abolished by transformation to the autolysin-proficient phenotype. The original Tupelo strain was discovered to be a mixture: a strain showing a vancomycin-tolerant phenotype (Tupelo_VT) and a vancomycin-nontolerant strain (Tupelo_VNT). The two strains differed only in terms of a single mutation in the ciaH gene present in the VT strain. Most interestingly, although the vancomycin tolerance of Tupelo_VT could be overcome by increasing the LytA dosage upon transformation by a multicopy plasmid or by externally adding the autolysin, we show that vancomycin tolerance in S. pneumoniae requires the simultaneous presence of a mutated CiaH histidine kinase and capsular polysaccharide.
Moscoso
Miriam
M
Centro de Investigaciones Biológicas (CSIC) and Centro de Investigación Biomédica en Red de Enfermedades Respiratorias (CIBERES), Ramiro de Maeztu, 9, 28040 Madrid, Spain.
Domenech
Mirian
M
Centro de Investigaciones Biológicas (CSIC) and Centro de Investigación Biomédica en Red de Enfermedades Respiratorias (CIBERES), Ramiro de Maeztu, 9, 28040 Madrid, Spain.
García
Ernesto
E
Centro de Investigaciones Biológicas (CSIC) and Centro de Investigación Biomédica en Red de Enfermedades Respiratorias (CIBERES), Ramiro de Maeztu, 9, 28040 Madrid, Spain.
D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004361:Drug Tolerance; D016368:Frameshift Mutation; D018628:Gene Dosage; D005816:Genetic Complementation Test; D000071677:Histidine Kinase; D008969:Molecular Sequence Data; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000235:genetics / Q000378:metabolism*; D011135:Polysaccharides, Bacterial / Q000378:metabolism*; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D013290:Streptococcal Infections / Q000382:microbiology; D013296:Streptococcus pneumoniae / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D014640:Vancomycin / Q000494:pharmacology*
D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D011135:Polysaccharides, Bacterial; D014640:Vancomycin; D011494:Protein Kinases; D000071677:Histidine Kinase; D009238:N-Acetylmuramoyl-L-alanine Amidase