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Infect Immun, February 1998, p. 567-572, Vol. 66, No. 2
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Identification and Characterization of the PutP Proline Permease That Contributes to In Vivo Survival of Staphylococcus aureus in Animal Models

William R. Schwan,1,* Silvija N. Coulter,1 Eva Y. W. Ng,1 Michael H. Langhorne,1 Heather D. Ritchie,1 Linnea L. Brody,1 Shannon Westbrock-Wadman,1 Arnold S. Bayer,2 Kim R. Folger,1 and C. Kendall Stover1

PathoGenesis Corporation, Seattle, Washington 98119,1 and Division of Infectious Diseases and the St. John's Cardiovascular Research Center, Harbor- UCLA Medical Center, Torrance, California 905092

Received 22 September 1997/Returned for modification 30 October 1997/Accepted 18 November 1997

Staphylococcus aureus is an important pathogen of humans and other animals, causing bacteremia, abscesses, endocarditis, and other infectious syndromes. A signature-tagged mutagenesis (STM) system was adapted for use in studying the genes required for in vivo survival of S. aureus. An STM library was ultimately created in S. aureus RN6390, with Tn917 being used to create the transposon mutations. Pools of S. aureus RN6390 mutants were screened in mouse abscess, bacteremia, and wound infection models for growth attenuation after in vivo passage. One of the mutants that was identified displayed marked attenuation following large-pool screening in all three animal models, which was confirmed in bacteremia and endocarditis models of infection with a smaller pool of mutants. Sequence analysis of the entire open reading frame showed a 99% identity to the high-affinity proline permease (putP) gene characterized in another strain of S. aureus. In wound and murine abscess infection models, the putP mutant was approximately 10-fold more attenuated than was wild-type strain RN6390. Another S. aureus strain transduced with the putP mutation also displayed an attenuated phenotype after passage in the wound model. A [3H]proline uptake assay showed that less proline was specifically transported into the putP mutant than into strain RN6390. The reduced viability of the bacteria possessing the mutation in the S. aureus high-affinity proline permease suggests that proline scavenging by the bacteria is important for in vivo growth and proliferation and that analogs of proline may serve as potential antistaphylococcal therapeutic agents.

* Corresponding author. Mailing address: PathoGenesis Corp., 201 Elliott Ave. West, Seattle, WA 98119. Phone: (206) 467-8100. Fax: (206) 282-5065. E-mail: bschwan{at}path.path.com.




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