PIROTH L, QUE Y, PIU S, ENTENZA JM, MOREILLON P; Interscience Conference on Antimicrobial Agents and Chemotherapy (42nd : 2002 : San Diego, Calif.).
Abstr Intersci Conf Antimicrob Agents Chemother Intersci Conf Antimicrob Agents Chemother. 2002 Sep 27-30; 42: abstract no. B-275.
CHUV, Lausanne, Switzerland
BACKGROUND: Expression of the S. aureus fibronectin-binding protein A (FnBPA) in Lactococcus lactis indicated that FnBPA was sufficient for infectivity in EE. FnBPA is an adhesin composed of a N-terminal domain (A) suspected to bind Fg, three Fn-binding (B,C,D) domain and a C-terminal wall-anchoring (LPXTG) domain. The pathogenic role of these domains was tested in vitro and in vivo. METHODS: The fnbA gene from S. aureus 8325-4 was used as a template. Truncated genes containing various domains bracketed by the 5' ribosome-binding site and leader sequence, and the 3' LPXTG domain were generated by a PCR-ligation technique. These genes were subcloned in the lactococcal expression vector pOri23, and electroporated into L. lactis. Recombinants were tested for adherence to Fg and Fn, and for their ability to infect rats with EE. RESULTS: Conclusion: In vitro adherence tests confirmed the existence of several individual binding domains mediating attachment to Fg (A) and/or Fn (A,B,C,D). In vivo, the double Fg/Fb binding domain A was necessary and sufficient to induce EE. However, combining domains A and BCD, as in the native protein, increased infectivity by 100x. FnPBA is a multifunctional adhesin that offers an example of domain cooperation in pathogenesis.
Publication Types:
Keywords:
- Adhesins, Bacterial
- Animals
- Base Sequence
- Binding Sites
- Communicable Diseases
- Endocarditis
- Endocarditis, Bacterial
- Fibrinogen
- Fibronectins
- In Vitro
- Infection
- Lactococcus lactis
- Rats
- Staphylococcus aureus
- genetics
- immunology
- metabolism
- pharmacokinetics
Other ID:
UI: 102266779
From Meeting Abstracts