Identification of potential cell-surface proteins in Candida albicans and investigation of the role of a putative cell-surface glycosidase in adhesion and virulence.
Cell-surface proteins are attractive targets for the development of novel antifungals as they are more accessible to drugs than are intracellular targets. By using a computational biology approach, we identified 180 potential cell-surface proteins in Candida albicans, including the known cell-surface adhesin Als1 and other cell-surface antigens, such as Pra1 and Csa1. Six proteins (named Csf1-6 for cell-surface factors) were selected for further biological characterization. First, we verified that the selected CSF genes are expressed in the yeast and/or hyphal form and then we investigated the effect of the loss of each CSF gene on cell-wall integrity, filamentation, adhesion to mammalian cells and virulence. As a result, we identified Csf4, a putative glycosidase with an apparent orthologue in Saccharomyces cerevisiae (Utr2), as an important factor for cell-wall integrity and maintenance. Interestingly, deletion of CSF4 also resulted in a defect in filamentation, a reduction in adherence to mammalian cells in an in vitro adhesion assay, and a prolongation of survival in an immunocompetent mouse model of disseminated candidiasis. A delay in colonization of key organs (e.g. kidney) was also observed, which is consistent with a reduction in virulence of the csf4-deletion strain. These data indicate a key role for extracellular glycosidases in fungal pathogenesis and represent a new site for therapeutic intervention to cure and prevent fungal disease.
Alberti-Segui
Christine
C
Genome Therapeutics Corporation, Waltham, MA 02453, USA.
Morales
Arturo J
AJ
Xing
Heming
H
Kessler
Marco M
MM
Willins
Debra Aker
DA
Weinstock
Keith G
KG
Cottarel
Guillaume
G
Fechtel
Kim
K
Rogers
Bruce
B
D000595:Amino Acid Sequence; D000818:Animals; D002176:Candida albicans / Q000201:enzymology / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology*; D002448:Cell Adhesion / Q000502:physiology*; D002473:Cell Wall / Q000737:chemistry* / Q000201:enzymology; D017353:Gene Deletion; D005838:Genotype; D006026:Glycoside Hydrolases / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008322:Mammals; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010641:Phenotype; D020543:Proteome; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014774:Virulence
D008565:Membrane Proteins; D020543:Proteome; D006026:Glycoside Hydrolases