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Infection and Immunity, December 2000, p. 6871-6878, Vol. 68, No. 12
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Heterologously Expressed Staphylococcus aureus
Fibronectin-Binding Proteins Are Sufficient for Invasion of
Host Cells
Bhanu
Sinha,1,2,*
Patrice
Francois,2
Yok-Ai
Que,3
Muzaffar
Hussain,1
Christine
Heilmann,1
Philippe
Moreillon,3
Daniel
Lew,2
Karl-Heinz
Krause,4
Georg
Peters,1 and
Mathias
Herrmann1
Institute of Medical Microbiology, University
of Münster, D-48129 Münster,
Germany,1 and Division of Infectious
Diseases2 and Laboratory of
Aging,4 University Hospital of Geneva, CH-1211
Geneva 14, and Laboratory of Infectious Diseases,
University Hospital of Lausanne, CH-1011
Lausanne,3 Switzerland
Received 26 July 2000/Returned for modification 25 August
2000/Accepted 6 September 2000
Staphylococcus aureus invasion of mammalian cells,
including epithelial, endothelial, and fibroblastic cells, critically
depends on fibronectin bridging between S. aureus
fibronectin-binding proteins (FnBPs) and the host fibronectin receptor
integrin
5
1 (B. Sinha et al., Cell.
Microbiol. 1:101-117, 1999). However, it is unknown whether this
mechanism is sufficient for S. aureus invasion. To address
this question, various S. aureus adhesins (FnBPA, FnBPB,
and clumping factor [ClfA]) were expressed in Staphylococcus carnosus and Lactococcus lactis subsp.
cremoris. Both noninvasive gram-positive microorganisms are
genetically distinct from S. aureus, lack any known
S. aureus surface protein, and do not bind fibronectin.
Transformants of S. carnosus and L. lactis
harboring plasmids coding for various S. aureus surface
proteins (FnBPA, FnBPB, and ClfA) functionally expressed adhesins (as
determined by bacterial clumping in plasma, specific latex
agglutination, Western ligand blotting, and binding to immobilized and
soluble fibronectin). FnBPA or FnBPB but not of ClfA conferred
invasiveness to S. carnosus and L. lactis.
Invasion of 293 cells by transformants was comparable to that of
strongly invasive S. aureus strain Cowan 1. Binding of
soluble and immobilized fibronectin paralleled invasiveness, demonstrating that the amount of accessible surface FnBPs is rate limiting. Thus, S. aureus FnBPs confer invasiveness to
noninvasive, apathogenic gram-positive cocci. Furthermore, FnBP-coated
polystyrene beads were internalized by 293 cells, demonstrating that
FnBPs are sufficient for invasion of host cells without the need for (S. aureus-specific) coreceptors.
*
Corresponding author. Mailing address: Institute of
Medical Microbiology, University of Münster, Domagkstraße 10, D-48149 Münster, Germany. Phone: 49-251-83-553-45. Fax:
49-251-83-553-50. E-mail: Bhanu.Sinha{at}gmx.de.
Infection and Immunity, December 2000, p. 6871-6878, Vol. 68, No. 12
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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