title: Cloning and disruption of caPLB1, a phospholipase B gene involved in the pathogenicity of Candida albicans.
abstract: The Candida albicans PLB1 gene was cloned using a polymerase chain reaction-based approach relying on degenerate oligonucleotide primers designed according to the amino acid sequences of two peptide fragments obtained from a purified candidal enzyme displaying phospholipase activity (Mirbod, F., Banno, Y., Ghannoum, M. A., Ibrahim, A. S., Nakashima, S., Yasuo, K., Cole, G. T., and Nozawa, Y. (1995) Biochim. Biophys. Acta 1257, 181-188). Sequence analysis of a 6.7-kilobase pair EcoRI-ClaI genomic clone revealed a single open reading frame of 1818 base pairs that predicts for a pre-protein of 605 residues. Comparison of the putative candidal phospholipase with those of other proteins in data base revealed significant homology to known fungal phospholipase Bs from Saccharomyces cerevisiae (45%), Penicillium notatum (42%), Torulaspora delbrueckii (48%), and Schizosaccharomyces pombe (38%). Thus, we have cloned the gene encoding a C. albicans phospholipase B homolog. This gene, designated caPLB1, was mapped to chromosome 6. Disruption experiments revealed that the caplb1 null mutant is viable and displays no obvious phenotype. However, the virulence of strains deleted for caPLB1, as assessed in a murine model for hematogenously disseminated candidiasis, was significantly attenuated compared with the isogenic wild-type parental strain. Although deletion of caPLB1 did not produce any detectable effects on candidal adherence to human endothelial or epithelial cells, the ability of the caplb1 null mutant to penetrate host cells was dramatically reduced. Thus, phospholipase B may well contribute to the pathogenicity of C. albicans by abetting the fungus in damaging and traversing host cell membranes, processes which likely increase the rapidity of disseminated infection.
authors.affiliation: Center for Medical Mycology, University Hospitals of Cleveland, and Case Western Reserve University, Cleveland, Ohio 44106-5028, USA.
authors.forename: S D
authors.initials: SD
authors.lastname: Leidich
authors.forename: A S
authors.initials: AS
authors.lastname: Ibrahim
authors.forename: Y
authors.initials: Y
authors.lastname: Fu
authors.forename: A
authors.initials: A
authors.lastname: Koul
authors.forename: C
authors.initials: C
authors.lastname: Jessup
authors.forename: J
authors.initials: J
authors.lastname: Vitullo
authors.forename: W
authors.initials: W
authors.lastname: Fonzi
authors.forename: F
authors.initials: F
authors.lastname: Mirbod
authors.forename: S
authors.initials: S
authors.lastname: Nakashima
authors.forename: Y
authors.initials: Y
authors.lastname: Nozawa
authors.forename: M A
authors.initials: MA
authors.lastname: Ghannoum
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002176:Candida albicans / Q000472:pathogenicity*; D002177:Candidiasis / Q000382:microbiology; D002448:Cell Adhesion / Q000235:genetics; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004195:Disease Models, Animal; D005656:Fungal Proteins / Q000737:chemistry*; D018390:Gene Targeting / Q000379:methods; D007668:Kidney / Q000382:microbiology / Q000648:ultrastructure; D008245:Lysophospholipase / Q000235:genetics*; D008565:Membrane Proteins; D051379:Mice; D008807:Mice, Inbred BALB C; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D029701:Saccharomyces cerevisiae Proteins; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014774:Virulence / Q000235:genetics*
chemical_list: D005656:Fungal Proteins; D008565:Membrane Proteins; C490271:PLB1 protein, Candida albicans; D029701:Saccharomyces cerevisiae Proteins; D008245:Lysophospholipase; C490270:PLB1 protein, S cerevisiae