Glycerol Uptake Is Important for L-Form Formation and Persistence in Staphylococcus aureus

S. aureus is a significant human pathogen and has previously been shown to form cell wall deficient forms or L-forms in vitro and in vivo during infection. Despite many previous studies on S. aureus L-forms, the mechanisms of L-form formation in this organism remain unknown. Here we established the L-form model in S. aureus and constructed a transposon mutant library to identify genes involved in L-form formation. Screening of the library for mutants defective in L-form formation identified glpF involved in glycerol uptake being important for L-form formation in S. aureus. Consistent with this observation, glpF was found to be highly expressed in L-form S. aureus but hardly expressed in normal walled form. In addition, glpF mutant was found to be defective in antibiotic persistence. The defect in L-form formation and antibiotic persistence of the glpF mutant could be complemented by the wild type glpF gene. These findings provide new insight into the mechanisms of L-form formation and persistence in S. aureus and may have implications for development of new drugs targeting persisters for improved treatment.

[1]  Ying Zhang Persisters, persistent infections and the Yin–Yang model , 2014, Emerging Microbes & Infections.

[2]  R. Bertram,et al.  Staphylococcus aureus Persisters Tolerant to Bactericidal Antibiotics , 2012, Journal of Molecular Microbiology and Biotechnology.

[3]  J. Errington,et al.  The rod to L‐form transition of Bacillus subtilis is limited by a requirement for the protoplast to escape from the cell wall sacculus , 2012, Molecular microbiology.

[4]  K. Devine Bacterial L‐forms on tap: an improved methodology to generate Bacillus subtilis L‐forms heralds a new era of research , 2012, Molecular microbiology.

[5]  G. Domingue Demystifying pleomorphic forms in persistence and expression of disease: Are they bacteria, and is peptidoglycan the solution? , 2010, Discovery medicine.

[6]  W. Shi,et al.  Energy production genes sucB and ubiF are involved in persister survival and tolerance to multiple antibiotics and stresses in Escherichia coli. , 2010, FEMS microbiology letters.

[7]  Ying Zhang,et al.  Insights into the Molecular Basis of L-Form Formation and Survival in Escherichia coli , 2009, PloS one.

[8]  J. Errington,et al.  Life without a wall or division machine in Bacillus subtilis , 2009, Nature.

[9]  R. D'ari,et al.  Unstable Escherichia coli L Forms Revisited: Growth Requires Peptidoglycan Synthesis , 2007, Journal of bacteriology.

[10]  N. Markova,et al.  Persistence of Staphylococcus aureus L-form during experimental lung infection in rats. , 2007, FEMS microbiology letters.

[11]  M. Vulić,et al.  GlpD and PlsB Participate in Persister Cell Formation in Escherichia coli , 2006, Journal of bacteriology.

[12]  Matthias Platzer,et al.  The analysis of cell division and cell wall synthesis genes reveals mutationally inactivated ftsQ and mraY in a protoplast-type L-form of Escherichia coli. , 2006, FEMS microbiology letters.

[13]  S. Projan,et al.  Transcription Profiling of the mgrA Regulon in Staphylococcus aureus , 2006, Journal of bacteriology.

[14]  T. Fawcett,et al.  β-Lactam Resistance in Staphylococcus aureus Cells That Do Not Require a Cell Wall for Integrity , 2005, Antimicrobial Agents and Chemotherapy.

[15]  Y. Kasahara,et al.  Induction of L-form-like cell shape change of Bacillus subtilis under microculture conditions. , 2003, Microbiology.

[16]  Robert M Stroud,et al.  Glycerol facilitator GlpF and the associated aquaporin family of channels. , 2003, Current opinion in structural biology.

[17]  H. B. Woody,et al.  Bacterial persistence and expression of disease , 1997, Clinical microbiology reviews.

[18]  M. Hiura,et al.  [Methicillin-resistant Staphylococcus aureus forming the fried egg appearance colonies isolated from a patient with septicemia]. , 1995, Rinsho byori. The Japanese journal of clinical pathology.

[19]  W. E. Owens Isolation of Staphylococcus aureus L forms from experimentally induced bovine mastitis , 1987, Journal of clinical microbiology.

[20]  P. Sears,et al.  Isolation of L-form variants after antibiotic treatment in Staphylococcus aureus bovine mastitis. , 1987, Journal of the American Veterinary Medical Association.

[21]  R. Novick,et al.  Complete nucleotide sequence of pT181, a tetracycline-resistance plasmid from Staphylococcus aureus. , 1983, Plasmid.

[22]  S. Anghelescu [Bacterial L forms]. , 1980, Revista de igiena, bacteriologie, virusologie, parazitologie, epidemiologie, pneumoftiziologie. Bacteriologia, virusologia, parazitologia, epidemiologia.

[23]  R. Banville FACTORS AFFECTING GROWTH OF STAPHYLOCOCCUS AUREUS L FORMS ON SEMIDEFINED MEDIUM , 1964, Journal of bacteriology.

[24]  H. Morton,et al.  Staphylococcus aureus , 1948 .

[25]  Elizabeth M Glass,et al.  Generating a collection of insertion mutations in the Staphylococcus aureus genome using bursa aurealis. , 2008, Methods in molecular biology.

[26]  K. Lewis,et al.  Persister cells and tolerance to antimicrobials. , 2004, FEMS microbiology letters.

[27]  J. BOGOJEVIC-DJORDJEVIC ‘L Forms’ of Bacteria , 1957, Nature.

[28]  E. Klieneberger The natural occurrence of pleuropneumonia‐like organism in apparent symbiosis with Strrptobacillus moniliformis and other bacteria , 1935 .