Analysis of PRA1 and its relationship to Candida albicans- macrophage interactions.
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.
Marcil
A
A
Genetics Group, Biotechnology Research Institute, National Research Council of Canada, Montreal, Québec, Canada. anne.marcil@cnrc-nrc.gc.ca
Gadoury
C
C
Ash
J
J
Zhang
J
J
Nantel
A
A
Whiteway
M
M
D000818:Animals; D002176:Candida albicans / Q000276:immunology*; D002460:Cell Line; D002478:Cells, Cultured; D005340:Fibrinogen / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D020869:Gene Expression Profiling; D006801:Humans; D008264:Macrophages / Q000382:microbiology*; D051379:Mice; D016254:Mutagenesis, Insertional; D020411:Oligonucleotide Array Sequence Analysis; D010588:Phagosomes / Q000382:microbiology; D011485:Protein Binding
D005656:Fungal Proteins; C088491:PRA1 protein, Candida albicans; D005340:Fibrinogen