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Journal of Bacteriology, August 2005, p. 5318-5329, Vol. 187, No. 15
0021-9193/05/$08.00+0     doi:10.1128/JB.187.15.5318-5329.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Staphylococcus aureus Develops an Alternative, ica-Independent Biofilm in the Absence of the arlRS Two-Component System{dagger}

Alejandro Toledo-Arana,1 Nekane Merino,1 Marta Vergara-Irigaray,1 Michel Débarbouillé,2 José R. Penadés,3 and Iñigo Lasa1*

Laboratory of Microbial Biofilms, Instituto de Agrobiotecnología and Departamento de Producción Agraria, Universidad Pública de Navarra-CSIC, Pamplona-31006, Spain,1 Biologie des Bactéries pathogènes à Gram positif, Département de Microbiologie Fondamentale et Medícale (CNRS, URA 2172), Institut Pasteur, Paris 75724, France,2 Cardenal Herrera-CEU University and Instituto Valenciano de Investigaciones Agrarias (IVIA), 46113 Moncada, Valencia, Spain3

Received 18 March 2005/ Accepted 28 April 2005

The biofilm formation capacity of Staphylococcus aureus clinical isolates is considered an important virulence factor for the establishment of chronic infections. Environmental conditions affect the biofilm formation capacity of S. aureus, indicating the existence of positive and negative regulators of the process. The majority of the screening procedures for identifying genes involved in biofilm development have been focused on genes whose presence is essential for the process. In this report, we have used random transposon mutagenesis and systematic disruption of all S. aureus two-component systems to identify negative regulators of S. aureus biofilm development in a chemically defined medium (Hussain-Hastings-White modified medium [HHWm]). The results of both approaches coincided in that they identified arlRS as a repressor of biofilm development under both steady-state and flow conditions. The arlRS mutant exhibited an increased initial attachment as well as increased accumulation of poly-N-acetylglucosamine (PNAG). However, the biofilm formation of the arlRS mutant was not affected when the icaADBC operon was deleted, indicating that PNAG is not an essential compound of the biofilm matrix produced in HHWm. Disruption of the major autolysin gene, atl, did not produce any effect on the biofilm phenotype of an arlRS mutant. Epistatic experiments with global regulators involved in staphylococcal-biofilm formation indicated that sarA deletion abolished, whereas agr deletion reinforced, the biofilm development promoted by the arlRS mutation.

* Corresponding author. Mailing address: Laboratory of Microbial Biofilms, Instituto de Agrobiotecnología, Universidad Pública de Navarra, Pamplona-31006 Spain. Phone: 34 948 168007. Fax: 34 948 232191. E-mail: ilasa{at}unavarra.es.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

Journal of Bacteriology, August 2005, p. 5318-5329, Vol. 187, No. 15
0021-9193/05/$08.00+0     doi:10.1128/JB.187.15.5318-5329.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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