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Infection and Immunity, February 2005, p. 872-877, Vol. 73, No. 2
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.2.872-877.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Staphylococcus aureus Virulence Factors Identified by Using a High-Throughput Caenorhabditis elegans-Killing Model

Jakob Begun,1,2,{dagger} Costi D. Sifri,3,{dagger},{ddagger} Samuel Goldman,1 Stephen B. Calderwood,3,4 and Frederick M. Ausubel1,2*

Department of Molecular Biology,1 Division of Infectious Diseases, Massachusetts General Hospital,3 Department of Microbiology and Molecular Genetics,4 Department of Genetics, Harvard Medical School, Boston, Massachusetts2

Received 3 July 2004/ Returned for modification 31 August 2004/ Accepted 5 October 2004

Staphylococcus aureus is an important human pathogen that is also able to kill the model nematode Caenorhabditis elegans. We constructed a 2,950-member Tn917 transposon insertion library in S. aureus strain NCTC 8325. Twenty-one of these insertions exhibited attenuated C. elegans killing, and of these, 12 contained insertions in different genes or chromosomal locations. Ten of these 12 insertions showed attenuated killing phenotypes when transduced into two different S. aureus strains, and 5 of the 10 mutants correspond to genes that have not been previously identified in signature-tagged mutagenesis studies. These latter five mutants were tested in a murine renal abscess model, and one mutant harboring an insertion in nagD exhibited attenuated virulence. Interestingly, Tn917 was shown to have a very strong bias for insertions near the terminus of DNA replication.

* Corresponding author. Mailing address: Department of Molecular Biology, Massachusetts General Hospital, 50 Blossom St., Boston, MA 02114. Phone: (617) 726-5950. Fax: (617) 726-5949. E-mail: ausubel{at}molbio.mgh.harvard.edu.

Editor: A. D. O'Brien

{dagger} J.B. and C.D.S. contributed equally to this work.

{ddagger} Present address: Division of Infectious Diseases and International Health, University of Virginia Health System, Charlottesville, VA 22908.

Infection and Immunity, February 2005, p. 872-877, Vol. 73, No. 2
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.2.872-877.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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