Monitoring Phenotypic Switching in <i>Candida albicans</i> and the Use of Next‐Gen Fluorescence Reporters

Frazer, Corey; Hernday, Aaron D.; Bennett, Richard J.

doi:10.1002/cpmc.76

Cited by 16 publications

(13 citation statements)

References 65 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…At the colony level, switching is evident by darker opaque sectors within white colonies and can be readily detected by state-specific fluorescent reporters ( Fig. 1a , b ) 37 – 39 . The TRN regulating the white-opaque switch shows multiple parallels to those defining mammalian cell fate.…”

Section: Resultsmentioning

confidence: 99%

Epigenetic cell fate in Candida albicans is controlled by transcription factor condensates acting at super-enhancer-like elements

et al. 2020

Self Cite

View full text Add to dashboard Cite

Summary Cell identity in eukaryotes is controlled by transcriptional regulatory networks (TRNs) that define cell type-specific gene expression. In the opportunistic fungal pathogen Candida albicans , TRNs regulate epigenetic switching between two alternative cell states, ‘white’ and ‘opaque’, that exhibit distinct host interactions. Here, we reveal that the transcription factors (TFs) regulating cell identity contain prion-like domains (PrLDs) that enable liquid-liquid demixing and the formation of phase-separated condensates. Multiple white-opaque TFs can co-assemble into complex condensates as observed on single DNA molecules. Moreover, heterotypic interactions between PrLDs supports the assembly of multifactorial condensates at a synthetic locus within live eukaryotic cells. Mutation of the Wor1 PrLD revealed that substitution of acidic residues abolished its ability to phase separate and to co-recruit other TFs in live cells, as well as its function in C. albicans cell fate determination. Together, these studies reveal that PrLDs support the assembly of TF complexes that control fungal cell identity and highlight parallels with the ‘super-enhancers’ that regulate mammalian cell fate.

show abstract

Section: Resultsmentioning

confidence: 99%

Epigenetic cell fate in Candida albicans is controlled by transcription factor condensates acting at super-enhancer-like elements

et al. 2020

Self Cite

View full text Add to dashboard Cite

show abstract

“…The second set of C . albicans mutants was constructed in strain SC5314 using the transient CRISPR-Cas9 system [ 36 ] combined with the Nat flipper approach [ 38 , 39 ]. Briefly, the gRNA and Cas9 constructs were amplified from vector pV1093 ( S3 Table ) by PCR [ 40 ].…”

Section: Methodsmentioning

confidence: 99%

“…Briefly, the gRNA and Cas9 constructs were amplified from vector pV1093 (S3 Table ) by PCR [40]. The gene deletion constructs with the maltose promoter-driven flippase and actin promoter-driven clonNAT selective marker were amplified with primers carrying micro-arms that were homologous to the flanking regions of target genes from a vector derived from pSF2A-mScarlet [38]. The gRNA, Cas9, and repair constructs were mixed and transformed into C. albicans cells using the lithium acetate heat-shock method [41].…”

Section: Fungal Strainsmentioning

confidence: 99%

Activation of EphA2-EGFR signaling in oral epithelial cells by Candida albicans virulence factors

et al. 2021

View full text Add to dashboard Cite

During oropharyngeal candidiasis (OPC), Candida albicans invades and damages oral epithelial cells, which respond by producing proinflammatory mediators that recruit phagocytes to foci of infection. The ephrin type-A receptor 2 (EphA2) detects β-glucan and plays a central role in stimulating epithelial cells to release proinflammatory mediators during OPC. The epidermal growth factor receptor (EGFR) also interacts with C. albicans and is known to be activated by the Als3 adhesin/invasin and the candidalysin pore-forming toxin. Here, we investigated the interactions among EphA2, EGFR, Als3 and candidalysin during OPC. We found that EGFR and EphA2 constitutively associate with each other as part of a heteromeric physical complex and are mutually dependent for C. albicans-induced activation. Als3-mediated endocytosis of a C. albicans hypha leads to the formation of an endocytic vacuole where candidalysin accumulates at high concentration. Thus, Als3 potentiates targeting of candidalysin, and both Als3 and candidalysin are required for C. albicans to cause maximal damage to oral epithelial cells, sustain activation of EphA2 and EGFR, and stimulate pro-inflammatory cytokine and chemokine secretion. In the mouse model of OPC, C. albicans-induced production of CXCL1/KC and CCL20 is dependent on the presence of candidalysin and EGFR, but independent of Als3. The production of IL-1α and IL-17A also requires candidalysin but is independent of Als3 and EGFR. The production of TNFα requires Als1, Als3, and candidalysin. Collectively, these results delineate the complex interplay among host cell receptors EphA2 and EGFR and C. albicans virulence factors Als1, Als3 and candidalysin during the induction of OPC and the resulting oral inflammatory response.

show abstract

“…A CAP1 ‐mNeonGreen cassette was amplified from pRB895 ( pSFS2A‐mNeonGreen ) (Frazer et al , ) using oligonucleotides 4564 and 4565 (see Appendix Table S5), which contain homology to the CAP1 gene and the mNeonGreen plasmid. This cassette was transformed into the diploid strain SC5314 and the tetraploid strain RBY18 to create strains CAY9331 (diploid) and CAY9332 (tetraploid).…”

Section: Methodsmentioning

confidence: 99%

Metabolism‐induced oxidative stress and DNA damage selectively trigger genome instability in polyploid fungal cells

et al. 2019

View full text Add to dashboard Cite

Understanding how cellular activities impact genome stability is critical to multiple biological processes including tumorigenesis and reproductive biology. The fungal pathogen Candida albicans displays striking genome dynamics during its parasexual cycle as tetraploid cells, but not diploid cells, exhibit genome instability and reduce their ploidy when grown on a glucose‐rich “pre‐sporulation” medium. Here, we reveal that C. albicans tetraploid cells are metabolically hyperactive on this medium with higher rates of fermentation and oxidative respiration relative to diploid cells. This heightened metabolism results in elevated levels of reactive oxygen species (ROS), activation of the ROS‐responsive transcription factor Cap1, and the formation of DNA double‐strand breaks. Genetic or chemical suppression of ROS levels suppresses each of these phenotypes and also protects against genome instability. These studies reveal how endogenous metabolic processes can generate sufficient ROS to trigger genome instability in polyploid C. albicans cells. We also discuss potential parallels with metabolism‐induced instability in cancer cells and speculate that ROS‐induced DNA damage could have facilitated ploidy cycling prior to a conventional meiosis in eukaryotes.

show abstract

Monitoring Phenotypic Switching in Candida albicans and the Use of Next‐Gen Fluorescence Reporters

Cited by 16 publications

References 65 publications

Epigenetic cell fate in Candida albicans is controlled by transcription factor condensates acting at super-enhancer-like elements

Epigenetic cell fate in Candida albicans is controlled by transcription factor condensates acting at super-enhancer-like elements

Activation of EphA2-EGFR signaling in oral epithelial cells by Candida albicans virulence factors

Metabolism‐induced oxidative stress and DNA damage selectively trigger genome instability in polyploid fungal cells

Contact Info

Product

Resources

About