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1.
Eukaryot Cell ; 5(8): 1252-65, 2006 Aug.
Article in English | MEDLINE | ID: mdl-16896210

ABSTRACT

In Candida albicans, Myo5p and Sla2p are required for the polarized localization and function of cortical actin patches, for hyphal formation, and for endocytosis. Deletion of either the MYO5 or the SLA2 gene generated a common transcriptional response that involved changes in the transcript levels of cell wall protein- and membrane protein-encoding genes. However, these profiles were distinct from those observed for a mutant with specific deletions of the actin-organizing domains of Myo5p or for wild-type cells treated with cytochalasin A, both of which also generate defects in the organization of cortical actin patches. The profiles observed for the myo5Delta and sla2Delta mutants had similarities to those of wild-type cells subjected to an osmotic shock, and the defects in cortical patch function found with myo5Delta and sla2Delta mutants, but not cortical actin patch distribution per se, affected sensitivity to various stresses, including heat and osmotic shocks and cell wall damage. Secondary effects coupled with defective endocytosis, such as lack of polarized lipid rafts and associated protein Rvs167-GFP (where GFP is green fluorescent protein) and lack of polarized wall remodeling protein GFP-Gsc1, were also observed for the myo5Delta and sla2Delta mutants. The mitogen-activated protein kinases Hog1p and Mkc1p, which mediate signaling in response to osmotic stress and cell wall damage, do not play a major role in regulating the transcript level changes in the myo5Delta and sla2Delta mutants. Hog1p was not hyperphosphorylated in the myo5Delta and sla2Delta mutants, and the transcript levels of only a subset of genes affected in the myo5Delta mutant were dependent upon the presence of Hog1p and Mkc1p. However, it appears that Hog1p and Mkc1p play important roles in the myo5Delta mutant cells because double deletion of myosin I and either Hog1p or Mkc1p resulted in very-slow-growing cells.


Subject(s)
Actins/genetics , Candida albicans/genetics , Fungal Proteins/genetics , Gene Expression Profiling , Membrane Proteins/genetics , Signal Transduction/genetics , Actins/physiology , Base Sequence , Cell Membrane/genetics , Cell Membrane/physiology , Cell Wall/genetics , Cell Wall/physiology , Gene Deletion , Genome , Hot Temperature , Molecular Sequence Data , Osmotic Pressure
2.
PLoS Genet ; 1(1): 36-57, 2005 Jul.
Article in English | MEDLINE | ID: mdl-16103911

ABSTRACT

Recent sequencing and assembly of the genome for the fungal pathogen Candida albicans used simple automated procedures for the identification of putative genes. We have reviewed the entire assembly, both by hand and with additional bioinformatic resources, to accurately map and describe 6,354 genes and to identify 246 genes whose original database entries contained sequencing errors (or possibly mutations) that affect their reading frame. Comparison with other fungal genomes permitted the identification of numerous fungus-specific genes that might be targeted for antifungal therapy. We also observed that, compared to other fungi, the protein-coding sequences in the C. albicans genome are especially rich in short sequence repeats. Finally, our improved annotation permitted a detailed analysis of several multigene families, and comparative genomic studies showed that C. albicans has a far greater catabolic range, encoding respiratory Complex 1, several novel oxidoreductases and ketone body degrading enzymes, malonyl-CoA and enoyl-CoA carriers, several novel amino acid degrading enzymes, a variety of secreted catabolic lipases and proteases, and numerous transporters to assimilate the resulting nutrients. The results of these efforts will ensure that the Candida research community has uniform and comprehensive genomic information for medical research as well as for future diagnostic and therapeutic applications.

3.
Eukaryot Cell ; 3(5): 1272-86, 2004 Oct.
Article in English | MEDLINE | ID: mdl-15470256

ABSTRACT

The molecular motor myosin I is required for hyphal growth in the pathogenic yeast Candida albicans. Specific myosin I functions were investigated by a deletion analysis of five neck and tail regions. Hyphal formation requires both the TH1 region and the IQ motifs. The TH2 region is important for optimal hyphal growth. All of the regions, except for the SH3 and acidic (A) regions that were examined individually, were required for the localization of myosin I at the hyphal tip. Similarly, all of the domains were required for the association of myosin I with pelletable actin-bound complexes. Moreover, the hyphal tip localization of cortical actin patches, identified by both rhodamine-phalloidin staining and Arp3-green fluorescent protein signals, was dependent on myosin I. Double deletion of the A and SH3 domains depolarized the distribution of the cortical actin patches without affecting the ability of the mutant to form hyphae, suggesting that myosin I has distinct functions in these processes. Among the six myosin I tail domain mutants, the ability to form hyphae was strictly correlated with endocytosis. We propose that the uptake of cell wall remodeling enzymes and excess plasma membrane is critical for hyphal formation.


Subject(s)
Candida albicans/metabolism , Fungal Proteins/chemistry , Fungal Proteins/metabolism , Myosin Type I/chemistry , Myosin Type I/metabolism , Actins/genetics , Actins/metabolism , Amino Acid Motifs , Base Sequence , Candida albicans/genetics , Candida albicans/growth & development , Cell Wall/metabolism , DNA, Fungal/genetics , Endocytosis/genetics , Endocytosis/physiology , Fungal Proteins/genetics , Genes, Fungal , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Mutation , Myosin Type I/genetics , Open Reading Frames , Phenotype , Protein Structure, Tertiary , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Sequence Deletion
4.
Curr Opin Microbiol ; 7(4): 350-7, 2004 Aug.
Article in English | MEDLINE | ID: mdl-15358253

ABSTRACT

The human fungal pathogen Candida albicans has many morphological forms. Recent advances in genomics and cell biology are providing an improved understanding of the molecular regulation of cell shape, and providing insights into the relationships between morphogenesis and virulence. This understanding may improve our ability to develop strategies to combat Candida infections.


Subject(s)
Candida albicans/growth & development , Candida albicans/pathogenicity , Fungal Proteins/metabolism , Gene Expression Regulation, Fungal , Signal Transduction , Animals , Candida albicans/genetics , Candidiasis/microbiology , Female , Fungal Proteins/genetics , Humans , Mice , Morphogenesis , Virulence
5.
Can J Microbiol ; 49(8): 514-24, 2003 Aug.
Article in English | MEDLINE | ID: mdl-14608387

ABSTRACT

Culture medium affected the virulence of a strain of Candida albicans toward Galleria mellonella larvae, but the yeast growth rates in yeast extract - peptone - dextrose broth and synthetic Galleria serum were not correlated with yeast virulence. Virulent C. albicans grew rapidly in larval serum, whereas, it limited nodulation and continued development in vivo, producing toxins that damaged the hemocytes and fat body. Nonpathogenic yeast-phase cells grew slowly in larval serum but induced extensively melanized nodules in vivo and developed no further. There was no discernible relationship in 14 exo-enzymes between the virulent and avirulent yeast strains and virulence. The avirulent myosin-I-defective yeast cells were rapidly removed from the hemolymph in vivo because of lysozyme-mediated yeast agglutination and the possible binding of the yeast cells by lysozyme and apolipophorin-III. Both lysozyme and apolipophorin-III are proteins that bind beta-1,3-glucan. Finally, insects with nonpathogenic C. albicans exhibited induced immunity and were more resistant to candidiasis from the wild-type yeast cells than were noninduced insects.


Subject(s)
Candida albicans/genetics , Candida albicans/pathogenicity , Moths/microbiology , Animals , Apolipoproteins/metabolism , Candida albicans/growth & development , Candida albicans/immunology , Cell Adhesion , Culture Media , Enzymes/metabolism , Fungal Proteins/metabolism , Genes, Fungal , Hemocytes/microbiology , Hemolymph/microbiology , Immunization , Larva/growth & development , Larva/immunology , Larva/microbiology , Moths/growth & development , Moths/immunology , Muramidase/metabolism , Mutation , Virulence/genetics
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