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Infection and Immunity, September 2008, p. 4345-4358, Vol. 76, No. 9
0019-9567/08/$08.00+0     doi:10.1128/IAI.00588-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Analysis of PRA1 and Its Relationship to Candida albicans- Macrophage Interactions{triangledown} ,{dagger}

A. Marcil,* C. Gadoury, J. Ash, J. Zhang, A. Nantel, and M. Whiteway

Genetics Group, Biotechnology Research Institute, National Research Council of Canada, Montreal, Québec H4P 2R2, Canada

Received 23 April 2007/ Returned for modification 8 June 2007/ Accepted 28 June 2008

Phagocytosis of Candida albicans by either primary bone marrow-derived mouse macrophages or RAW 264.7 cells upregulated transcription of PRA1, which encodes a cell wall/membrane-associated antigen previously described as a fibrinogen binding protein. However, a pra1 null mutant was still able to bind fibrinogen, showing that Pra1p is not uniquely required for fibrinogen binding. As well, Pra1 tagged with green fluorescent protein did not colocalize with AlexaFluor 546-labeled human fibrinogen, and while PRA1 expression was inhibited when Candida was grown in fetal bovine serum-containing medium, Candida binding to fibrinogen was activated by these conditions. Therefore, it appears that Pra1p can play at most a minor role in fibrinogen binding to C. albicans. PRA1 gene expression is induced in vitro by alkaline pH, and therefore its activation in phagosomes suggested that phagosome maturation was suppressed by the presence of Candida cells. LysoTracker red-labeled organelles failed to fuse with phagosomes containing live Candida, while phagosomes containing dead Candida underwent a normal phagosome-to-phagolysosome maturation. Immunofluorescence staining with the early/recycling endosomal marker transferrin receptor (CD71) suggested that live Candida may escape macrophage destruction through the inhibition of phagolysosomal maturation.


* Corresponding author. Mailing address: Biotechnology Research Institute, National Research Council of Canada, 6100 Ave. Royalmount, Montreal, Québec H4P 2R2, Canada. Phone: (514) 496-1923. Fax: (514) 496-6213. E-mail: anne.marcil{at}cnrc-nrc.gc.ca

{triangledown} Published ahead of print on 14 July 2008.

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

Editor: A. Casadevall


Infection and Immunity, September 2008, p. 4345-4358, Vol. 76, No. 9
0019-9567/08/$08.00+0     doi:10.1128/IAI.00588-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.