Rapid Hypothesis Testing with
            <i>Candida albicans</i>
            through Gene Disruption with Short Homology Regions

Wilson, R. Bryce; Davis, Dana A.; Mitchell, Aaron P.

doi:10.1128/jb.181.6.1868-1874.1999

Cited by 738 publications

(354 citation statements)

References 19 publications

(15 reference statements)

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“…C. albicans BWP171CIp30 was used for all experiments (20). This isogenic wild-type strain was selected to facilitate future comparison with C. albicans deletion mutants.…”

Section: Experimental Methodsmentioning

confidence: 99%

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Epithelial invasion outcompetes hypha development during Candida albicans infection as revealed by an image‐based systems biology approach

et al. 2013

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Candida albicans is the most common opportunistic fungal pathogen of the human mucosal flora, frequently causing infections. The fungus is responsible for invasive infections in immunocompromised patients that can lead to sepsis. The yeast to hypha transition and invasion of host-tissue represent major determinants in the switch from benign colonizer to invasive pathogen. A comprehensive understanding of the infection process requires analyses at the quantitative level. Utilizing fluorescence microscopy with differential staining, we obtained images of C. albicans undergoing epithelial invasion during a time course of 6 h. An image-based systems biology approach, combining image analysis and mathematical modeling, was applied to quantify the kinetics of hyphae development, hyphal elongation, and epithelial invasion. The automated image analysis facilitates high-throughput screening and provided quantities that allow for the time-resolved characterization of the morphological and invasive state of fungal cells. The interpretation of these data was supported by two mathematical models, a kinetic growth model and a kinetic transition model, that were developed using differential equations. The kinetic growth model describes the increase in hyphal length and revealed that hyphae undergo mass invasion of epithelial cells following primary hypha formation. We also provide evidence that epithelial cells stimulate the production of secondary hyphae by C. albicans. Based on the kinetic transition model, the route of invasion was quantified in the state space of non-invasive and invasive fungal cells depending on their number of hyphae. This analysis revealed that the initiation of hyphae formation represents an ultimate commitment to invasive growth and suggests that in vivo, the yeast to hypha transition must be under exquisitely tight negative regulation to avoid the transition from commensal to pathogen invading the epithelium. V C 2013 International Society for Advancement of Cytometry

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“…C. albicans BWP171CIp30 was used for all experiments (20). This isogenic wild-type strain was selected to facilitate future comparison with C. albicans deletion mutants.…”

Section: Experimental Methodsmentioning

confidence: 99%

“…This suggests that the observed progression of length was predominantly induced by transitions of hyphae from the non-invasive state to the invasive state, such that the net length of invaded hyphae increased at a higher overall rate [see Supporting Information S3 Eq. (20)].…”

Section: Hyphal Growth Kinetics Reveals Massive Epithelial Invasionmentioning

confidence: 99%

Epithelial invasion outcompetes hypha development during Candida albicans infection as revealed by an image‐based systems biology approach

et al. 2013

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“…All C. albicans strains were derivatives of strain SC5314 (Gillum et al, 1984). Strain BWP17 (Wilson et al, 1999) was used to create all mutant strains. The genotypes of all strains constructed in this study are listed in Table 3.…”

Section: Strain Constructionmentioning

confidence: 99%

In vivo transcript profiling of Candida albicans identifies a gene essential for interepithelial dissemination

Zakikhany¹,

Naglik²,

Schmidt-Westhausen³

et al. 2007

Cellular Microbiology

268

334

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SummaryCandida albicans is the most common oral fungal pathogen of humans, but the mechanisms by which C. albicans invades and persists within mucosal epithelium are not clear. To understand oral pathogenesis, we characterized the cellular and molecular mechanisms of epithelial-fungus interactions using reconstituted human oral epithelium (RHE). We observed that hyphal formation facilitates epithelial invasion via both active (physical penetration) and passive (induced endocytosis) processes. Genome wide transcript profiling of C. albicans experimental RHE infection was compared with that from 11 patient samples with pseudomembranous candidiasis to identify genes associated with disease development in vivo. Expression profiles reflected the morphological switch and an adaptive response to neutral pH, non-glucose carbon sources and nitrosative stress. We identified several novel infection-associated genes with unknown function. One gene, upregulated in both RHE infection and patients, named EED1, was essential for maintenance of hyphal elongation. Mutants lacking EED1 showed transient cell elongation on epithelial tissue, which enabled only superficial invasion of epithelial cells. Once inside an epithelial cell, Deed1 cells could proliferate as yeasts or pseudohyphae but remained trapped intracellularly. Our results suggest that the adaptive response and morphology of C. albicans play specific roles for host-fungal interactions during mucosal infections.

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“…All C. albicans strains used in this study were derived from BWP17 (Wilson et al, 1999) and are listed in Table 2. Primer sequences from 5′ to 3′ are listed in Supporting Information, 'Primer sequences' worksheet.…”

Section: Construction Of Fungal Strainsmentioning

confidence: 99%

Candida albicanstranscription factor Rim101 mediates pathogenic interactions through cell wall functions

Nobile

Solis

Myers

et al. 2008

Cellular Microbiology

Self Cite

142

135

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SummarypH-responsive transcription factors of the Rim101/ PacC family govern virulence in many fungal pathogens. These family members control expression of target genes with diverse functions in growth, morphology and environmental adaptation, so the mechanistic relationship between Rim101/PacC and infection is unclear. We have focused on Rim101 from Candida albicans, which we find to be required for virulence in an oropharyngeal candidiasis model. Rim101 affects the yeast-hypha morphological transition, a major virulence requirement in disseminated infection models. However, virulence in the oropharyngeal candidiasis model is independent of the yeast-hypha transition because it is unaffected by an nrg1 mutation, which prevents formation of yeast cells. Here we have identified Rim101 target genes in an nrg1D/D mutant background and surveyed function using an overexpression-rescue approach. Increased expression of Rim101 target genes ALS3, CHT2, PGA7/RBT6, SKN1 or ZRT1 can partially restore pathogenic interaction of a rim101D/D mutant with oral epithelial cells. Four of these five genes govern cell wall structure. Our results indicate that Rim101-dependent cell wall alteration contributes to C. albicans pathogenic interactions with oral epithelial cells, independently of cell morphology.

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Rapid Hypothesis Testing with Candida albicans through Gene Disruption with Short Homology Regions

Cited by 738 publications

References 19 publications

Epithelial invasion outcompetes hypha development during Candida albicans infection as revealed by an image‐based systems biology approach

Epithelial invasion outcompetes hypha development during Candida albicans infection as revealed by an image‐based systems biology approach

In vivo transcript profiling of Candida albicans identifies a gene essential for interepithelial dissemination

Candida albicanstranscription factor Rim101 mediates pathogenic interactions through cell wall functions

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