Truncation of fibronectin-binding proteins in Staphylococcus aureus strain Newman leads to deficient adherence and host cell invasion due to loss of the cell wall anchor function.
Staphylococcus aureus fibronectin-binding proteins (FnBPs) play a critical role in S. aureus pathogenesis. FnBPs mediate adhesion to fibronectin and invasion of mammalian cells, including epithelial, endothelial, and fibroblastic cells, by fibronectin bridging to the host cell fibronectin receptor integrin (alpha(5))beta(1). Strain Newman is a laboratory strain frequently used for genetic, functional, and in vivo studies. However, despite pronounced production of FnBPs, strain Newman is only weakly adherent to immobilized Fn and weakly invasive. We examined whether these effects are due to a structural difference of FnBPs. Here, we show that both fnbA(Newman) and fnbB(Newman) contain a centrally located point mutation resulting in a stop codon. This leads to a truncation of both FnBPs at the end of the C domain at identical positions. Most likely, the stop codon occurred first in fnbB(Newman) and was subsequently transferred to fnbA(Newman) by replacement of the entire region encompassing the C, D, and W domains with the respective sequence of fnbB(Newman). Using heterologous expression in Staphylococcus carnosus, we found that truncated FnBPs were completely secreted into the culture medium and not anchored to the cell wall, since they lack the sortase motif (LPETG). Consequently, this led to a loss of FnBP-dependent functions, such as strong adhesion to immobilized fibronectin, binding of fibrinogen, and host cell invasion. This mutation may explain some of the earlier reported conflicting data with strain Newman. Thus, care should be taken when drawing negative conclusions about the role of FnBPs as a virulence factor in a given model.
Grundmeier
Matthias
M
Institute of Medical Microbiology, University of Münster, Domagkstrasse 10, D-48149 Münster, Germany.
Hussain
Muzaffar
M
Becker
Petra
P
Heilmann
Christine
C
Peters
Georg
G
Sinha
Bhanu
B
D018829:Adhesins, Bacterial / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D000595:Amino Acid Sequence; D001422:Bacterial Adhesion; D001483:Base Sequence; D002473:Cell Wall / Q000502:physiology*; D002478:Cells, Cultured; D018388:Codon, Terminator; D004697:Endocarditis, Bacterial / Q000209:etiology; D005353:Fibronectins / Q000502:physiology; D006801:Humans; D008969:Molecular Sequence Data; D013211:Staphylococcus aureus / Q000472:pathogenicity*
D018829:Adhesins, Bacterial; D018388:Codon, Terminator; D005353:Fibronectins; C078635:fibronectin-binding proteins, bacterial