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Infection and Immunity, September 2009, p. 3626-3638, Vol. 77, No. 9
0019-9567/09/$08.00+0     doi:10.1128/IAI.00219-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Contribution of Autolysin and Sortase A during Enterococcus faecalis DNA-Dependent Biofilm Development{triangledown} ,{dagger}

Pascale S. Guiton, Chia S. Hung, Kimberly A. Kline, Robyn Roth, Andrew L. Kau, Ericka Hayes, John Heuser, Karen W. Dodson, Michael G. Caparon,* and Scott J. Hultgren*

Department of Molecular Microbiology and Microbial Pathogenesis, Washington University in St. Louis, St. Louis, Missouri 63110

Received 25 February 2009/ Returned for modification 3 April 2009/ Accepted 7 June 2009

Biofilm production is a major attribute of Enterococcus faecalis clinical isolates. Although some factors, such as sortases, autolysin, and extracellular DNA (eDNA), have been associated with E. faecalis biofilm production, the mechanisms underlying the contributions of these factors to this process have not been completely elucidated yet. In this study we define important roles for the major E. faecalis autolysin (Atn), eDNA, and sortase A (SrtA) during the developmental stages of biofilm formation under static and hydrodynamic conditions. Deletion of srtA affects the attachment stage and results in a deficiency in biofilm production. Atn-deficient mutants are delayed in biofilm development due to defects in primary adherence and DNA release, which we show to be particularly important during the accumulative phase for maturation and architectural stability of biofilms. Confocal laser scanning and freeze-dry electron microscopy of biofilms grown under hydrodynamic conditions revealed that E. faecalis produces a DNase I-sensitive fibrous network, which is important for biofilm stability and is absent in atn-deficient mutant biofilms. This study establishes the stage-specific requirements for SrtA and Atn and demonstrates a role for Atn in the pathway leading to DNA release during biofilm development in E. faecalis.

* Corresponding author. Mailing address: Department of Molecular Microbiology and Microbial Pathogenesis, Washington University School of Medicine, 600 S. Euclid Ave., Campus Box 8230, Saint Louis, MO 63110-1093. Fax: (314) 362-1998. Phone for Scott J. Hultgren: (314) 362-6772. E-mail: hultgren{at}borcim.wustl.edu. Phone for Michael G. Caparon: (314) 362-1485. E-mail: caparon{at}borcim.wustl.edu

{triangledown} Published ahead of print on 15 June 2009.

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

Editor: A. Camilli

Infection and Immunity, September 2009, p. 3626-3638, Vol. 77, No. 9
0019-9567/09/$08.00+0     doi:10.1128/IAI.00219-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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