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Research Paper |
Protein Design Labs, Inc., 34801 Campus Drive, Fremont, CA 94555, USA1
Author for correspondence: Andrea Marra. Tel: +1 860 686 1507. Fax: +1 860 441 6159. e-mail: andrea_marra{at}groton.pfizer.com
Differential fluorescence induction technology was used to identify promoters of Streptococcus pneumoniae genes that are expressed during lung infection of the mouse. Among the promoter clones that were identified multiple times was the psa promoter, which drives expression of the psaBCA operon. These genes have been identified previously and shown to encode a manganese permease system as well as play a role in the virulence of this organism. Mutations in psaB, psaC or psaA result in growth limitation in low manganese. The expression of the psa operon was examined in vivo and the virulence of deletion mutants of psaB, psaC, psaA and psaBCA was assessed in four different animal models of infection. The psa promoter was induced more than ten-fold in vivo using an intraperitoneal chamber implant model. The psaB, psaC and psaA mutants were completely attenuated in systemic, respiratory tract and otitis media infections. In addition, these mutants were unable to grow in an implanted peritoneal chamber, but growth was restored by the addition of manganese to the chambers.
Keywords: manganese permease system, pneumococcal virulence
Abbreviations: DFI, differential fluorescence induction; FACS, fluorescence-activated cell sorting; GFP, green fluorescent protein; m.c.f., mean channel flourescence; RTI, respiratory tract infection; spc, spectinomycin
a Present address: Pfizer Global Research and Development, Antibacterials Discovery MS8118W-249, Eastern Point Road, Groton, CT 06340, USA.
b Present address: Genencor International, Inc., 925 Page Mill Road, Palo Alto, CA 94304, USA.
c Present address: Pharmacia Corporation, 7000 Portage Road, 6609–209–737, Kalamazoo, MI 49001-0199, USA.
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