Mapping and characterization of multiple chromosomal factors involved in methicillin resistance in Staphylococcus aureus.
Chromosomal factors, termed fem or aux factors, are needed for the expression of methicillin resistance in methicillin-resistant (Mcr) Staphylococcus aureus; also needed is the mec-encoded low-affinity penicillin-binding protein PBP 2'. These factors make up part of the normal set of genes present in susceptible and resistant strains of S. aureus and can be identified by Tn551-mediated insertional inactivation of the methicillin resistance. In this study, we characterized different Tn551 inserts and mapped them into four distinct loci on the SmaI chromosomal map of S. aureus NCTC 8325, thereby identifying two new loci which code for fem factors. The largest fragment, SmaI-A, carries three loci, two coding for both closely linked factors femA and femB and a novel third locus (femC) that is not linked to the other two. An additional, fourth, locus, femD, was identified in fragment SmaI-I. femA and femB inactivation reduced overall methicillin resistance, whereby femB had less of an influence on the resistance level. femC and femD inactivation reduced mainly the basal resistance level in heterogeneously Mcr strains and had less of an impact on the subpopulation with high-level resistance. Inactivation of either of these factors was shown to have no influence on the production of PBP 2', the main factor mediating methicillin resistance. In addition, no changes were observed in the banding patterns of the major autolysins in whole-cell extracts of the fem mutants, suggesting that the reduced cell wall turnover and autolysis observed in some of the insertionally inactivated strains were due to changes either of the substrate or in the autolysin control.
Berger-Bächi
B
B
Institute of Medical Microbiology, University of Zürich, Switzerland.
Strässle
A
A
Gustafson
J E
JE
Kayser
F H
FH
D001329:Autolysis; D001345:Autoradiography; D002473:Cell Wall / Q000378:metabolism; D002876:Chromosomes, Bacterial / Q000378:metabolism*; D005819:Genetic Markers; D016106:Methicillin Resistance / Q000235:genetics*; D013211:Staphylococcus aureus / Q000235:genetics* / Q000648:ultrastructure
D005819:Genetic Markers