Influence of femB on methicillin resistance and peptidoglycan metabolism in Staphylococcus aureus.
The inactivation of FemB by insertion of Tn551 in the central part of the femB open reading frame was shown to increase susceptibility of methicillin-resistant Staphylococcus aureus strains toward beta-lactam antibiotics to the same extent as did inactivation of femA. Strains carrying the methicillin resistance determinant (mec) and expressing PBP 2' were affected to the same extent as were strains selected for in vitro resistance, which did not express PBP 2'. Both femA and femB, which form an operon, are involved in a yet unknown manner in the glycine interpeptide bridge formation of the S. aureus peptidoglycan. FemB inactivation was shown to reduce the glycine content of peptidoglycan by approximately 40%, depending on the S. aureus strain. The reduction of the interpeptide bridge glycine content led to significant reduction in peptidoglycan cross-linking, as measured by gel permeation high-pressure liquid chromatography of muramidase-digested cell walls. Maximum peptide chain length was reduced by approximately 40%. It is shown that the complete pentaglycine interpeptide bridge is important for the sensitivity against beta-lactam antibiotics and for the undisturbed activity of the staphylococcal cell wall-synthesizing and hydrolyzing enzymes, as was also apparent from electron microscopic examinations, which revealed aberrant placement of cross walls and retarded cell separation, leading to a pseudomulticellular phenotype of the cells for both femA and femB mutants.
Henze
U
U
Institute of Medical Microbiology, University of Zürich, Switzerland.
Sidow
T
T
Wecke
J
J
Labischinski
H
H
Berger-Bächi
B
B
D000596:Amino Acids / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D004251:DNA Transposable Elements; D005798:Genes, Bacterial; D016106:Methicillin Resistance / Q000235:genetics*; D008854:Microscopy, Electron; D010457:Peptidoglycan / Q000737:chemistry / Q000378:metabolism*; D015183:Restriction Mapping; D013211:Staphylococcus aureus / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure
D000596:Amino Acids; D004251:DNA Transposable Elements; D010457:Peptidoglycan