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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 alpha 5beta 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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