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Infection and Immunity, May 2002, p. 2399-2407, Vol. 70, No. 5
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.5.2399-2407.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Conservation, Surface Exposure, and In Vivo Expression of the Frp Family of Iron-Regulated Cell Wall Proteins in Staphylococcus aureus

Julie A. Morrissey,1,2* Alan Cockayne,1 Jane Hammacott,1 Keith Bishop,1 Amy Denman-Johnson,1 Philip J. Hill,1,3 and Paul Williams1,4

Institute of Infections and Immunity, Queen's Medical Centre, University of Nottingham, Nottingham NG7 2UH,1 Department of Microbiology & Immunology, University of Leicester, Leicester LE1 9HN,2 School of Biosciences, Sutton Bonnington Campus, University of Nottingham, Loughborough, Leicestershire LE12 5RD,3 School of Pharmaceutical Sciences, University of Nottingham, Nottingham NG7 2RD, United Kingdom4

Received 8 November 2001/ Returned for modification 25 January 2002/ Accepted 11 February 2002

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis identified two conserved, immunogenic Staphylococcus aureus cell wall proteins, of 40 and 87 kDa, expressed under iron-restricted growth conditions in vitro and in vivo. N-terminal sequencing and subsequent genome analysis showed that these proteins are encoded by adjacent monocistronic open reading frames designated frpA and frpB, respectively. Studies with an S. aureus fur mutant confirmed that expression of FrpA and FrpB is regulated by Fur but that there also appears to be differential expression of these proteins in different iron-restricted media in vitro. FrpA and FrpB share some amino acid sequence homology with each other and with a putative S. aureus membrane protein, FrpC. frpC is the first gene of a Fur-regulated operon encoding four proteins of unknown function (FrpC, -D, -G, and -H) and the binding protein (FrpE) and permease (FrpF) of a putative iron transporter. Antisense mutagenesis and bioassays showed that FrpA and FrpB are not required for growth of S. aureus under iron-restricted conditions in vitro and do not appear to be involved in the transport of iron from siderophores or in binding of hemin. Further phenotypic analysis suggested that FrpA may be involved in adhesion of S. aureus to plastic in vitro. Binding of S. aureus to microtiter wells was found to be iron regulated, and iron-restricted S. aureus containing antisense frpA or frpAB but not frpB constructs showed reduced binding compared to vector construct controls.

* Corresponding author. Mailing address: Department of Microbiology & Immunology, University of Leicester, Medical Sciences Building, University Road, Leicester LE1 9HN, United Kingdom. Phone: 44-116-2522943. Fax: 44-116-2525030. E-mail: jam26{at}le.ac.uk.

Editor: E. I. Tuomanen

Infection and Immunity, May 2002, p. 2399-2407, Vol. 70, No. 5
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.5.2399-2407.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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