title: Living with an imperfect cell wall: compensation of femAB inactivation in Staphylococcus aureus.
abstract: BACKGROUND
  Synthesis of the Staphylococcus aureus peptidoglycan pentaglycine interpeptide bridge is catalyzed by the nonribosomal peptidyl transferases FemX, FemA and FemB. Inactivation of the femAB operon reduces the interpeptide to a monoglycine, leading to a poorly crosslinked peptidoglycan. femAB mutants show a reduced growth rate and are hypersusceptible to virtually all antibiotics, including methicillin, making FemAB a potential target to restore beta-lactam susceptibility in methicillin-resistant S. aureus (MRSA). Cis-complementation with wild type femAB only restores synthesis of the pentaglycine interpeptide and methicillin resistance, but the growth rate remains low. This study characterizes the adaptations that ensured survival of the cells after femAB inactivation.
  
  
  RESULTS
  In addition to slow growth, the cis-complemented femAB mutant showed temperature sensitivity and a higher methicillin resistance than the wild type. Transcriptional profiling paired with reporter metabolite analysis revealed multiple changes in the global transcriptome. A number of transporters for sugars, glycerol, and glycine betaine, some of which could serve as osmoprotectants, were upregulated. Striking differences were found in the transcription of several genes involved in nitrogen metabolism and the arginine-deiminase pathway, an alternative for ATP production. In addition, microarray data indicated enhanced expression of virulence factors that correlated with premature expression of the global regulators sae, sarA, and agr.
  
  
  CONCLUSIONS
  Survival under conditions preventing normal cell wall formation triggered complex adaptations that incurred a fitness cost, showing the remarkable flexibility of S. aureus to circumvent cell wall damage. Potential FemAB inhibitors would have to be used in combination with other antibiotics to prevent selection of resistant survivors.
authors.affiliation: Institute of Medical Microbiology, University of Zürich, Zürich, Switzerland. jhuebsch@immv.uzh.ch
authors.forename: Judith
authors.initials: J
authors.lastname: Hübscher
authors.forename: Andrea
authors.initials: A
authors.lastname: Jansen
authors.forename: Oliver
authors.initials: O
authors.lastname: Kotte
authors.forename: Juliane
authors.initials: J
authors.lastname: Schäfer
authors.forename: Paul A
authors.initials: PA
authors.lastname: Majcherczyk
authors.forename: Llinos G
authors.initials: LG
authors.lastname: Harris
authors.forename: Gabriele
authors.initials: G
authors.lastname: Bierbaum
authors.forename: Matthias
authors.initials: M
authors.lastname: Heinemann
authors.forename: Brigitte
authors.initials: B
authors.lastname: Berger-Bächi
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D002473:Cell Wall / Q000737:chemistry*; D019295:Computational Biology; D020868:Gene Silencing; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D009876:Operon; D011134:Polysaccharides / Q000737:chemistry; D013211:Staphylococcus aureus / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D013696:Temperature; D014158:Transcription, Genetic