Visualization of Cytokinesis Events in Budding Yeast by Transmission Electron Microscopy

Meitinger, Franz; Pereira, Gislene

doi:10.1007/978-1-4939-3145-3_7

Cited by 4 publications

(3 citation statements)

References 14 publications

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“…The most striking difference was the structure of the septum in dividing cells. In CDC14/CDC14 and CDC14/Δ cells the primary septum was uniformly visible as a bright, straight band bisecting the bud neck, with thin layers of secondary septum on either side ( Figures 5A , B ), as expected ( Meitinger and Pereira, 2016 ). Relatively normal primary septa were also observed in some cdc14 hm /Δ and cdc14Δ/Δ cells, however we frequently observed primary septa with abnormal morphologies in these strains that were never observed in wild-type cells ( Figures 5C , D ).…”

Section: Resultssupporting

confidence: 78%

Cdc14 phosphatase contributes to cell wall integrity and pathogenesis in Candida albicans

et al. 2023

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The Cdc14 phosphatase family is highly conserved in fungi. In Saccharomyces cerevisiae, Cdc14 is essential for down-regulation of cyclin-dependent kinase activity at mitotic exit. However, this essential function is not broadly conserved and requires only a small fraction of normal Cdc14 activity. Here, we identified an invariant motif in the disordered C-terminal tail of fungal Cdc14 enzymes that is required for full enzyme activity. Mutation of this motif reduced Cdc14 catalytic rate and provided a tool for studying the biological significance of high Cdc14 activity. A S. cerevisiae strain expressing the reduced-activity hypomorphic mutant allele (cdc14hm) as the sole source of Cdc14 proliferated like the wild-type parent strain but exhibited an unexpected sensitivity to cell wall stresses, including chitin-binding compounds and echinocandin antifungal drugs. Sensitivity to echinocandins was also observed in Schizosaccharomyces pombe and Candida albicans strains lacking CDC14, suggesting this phenotype reflects a novel and conserved function of Cdc14 orthologs in mediating fungal cell wall integrity. In C. albicans, the orthologous cdc14hm allele was sufficient to elicit echinocandin hypersensitivity and perturb cell wall integrity signaling. It also caused striking abnormalities in septum structure and the same cell separation and hyphal differentiation defects previously observed with cdc14 gene deletions. Since hyphal differentiation is important for C. albicans pathogenesis, we assessed the effect of reduced Cdc14 activity on virulence in Galleria mellonella and mouse models of invasive candidiasis. Partial reduction in Cdc14 activity via cdc14hm mutation severely impaired C. albicans virulence in both assays. Our results reveal that high Cdc14 activity is important for C. albicans cell wall integrity and pathogenesis and suggest that Cdc14 may be worth future exploration as an antifungal drug target.

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Section: Resultssupporting

confidence: 78%

Cdc14 phosphatase contributes to cell wall integrity and pathogenesis in Candida albicans

et al. 2023

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show abstract

“…The most striking difference was the structure of the septum in dividing cells. In CDC14/CDC14 and CDC14/Δ cells the primary septum was uniformly visible as a bright, straight band bisecting the bud neck, with thin layers of secondary septum on either side (Fig 5A-B), as expected (49). Relatively normal primary septa were also observed in some cdc14 hm /Δ and cdc14Δ/Δ cells, however we frequently observed primary septa with abnormal morphologies in these strains that were never observed in wild-type cells (Fig 5C-D).…”

Section: Cdc14 Deficiency Impairs C Albicans Septum Assembly and Cell...supporting

confidence: 81%

Reduced Cdc14 phosphatase activity impairs septation, hyphal differentiation and pathogenesis and causes echinocandin hypersensitivity inCandida albicans

Milholland

AbdelKhalek

Baker

et al. 2022

Preprint

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The Cdc14 phosphatase family is highly conserved in fungi. In Saccharomyces cerevisiae, Cdc14 is essential for down-regulation of cyclin-dependent kinase activity at mitotic exit. However, this essential function is not broadly conserved and requires a small fraction of normal Cdc14 activity. It remains unclear what fungal Cdc14 functions require high Cdc14 activity. We identified an invariant motif in the disordered C-terminal tail of fungal Cdc14 enzymes that is required for full enzyme activity. Mutation of this motif reduced Cdc14 catalytic rate and provided a tool for studying the biological significance of high Cdc14 activity. A S. cerevisiae strain expressing the reduced-activity hypomorphic mutant allele (cdc14hm) as the sole source of Cdc14 exhibited an unexpected sensitivity to cell wall stresses, including chitin-binding compounds and echinocandin antifungal drugs. Sensitivity to echinocandins was also observed in Schizosaccharomyces pombe and Candida albicans strains lacking CDC14, suggesting this phenotype reflects a conserved function of Cdc14 orthologs in mediating fungal cell wall integrity. In C. albicans, the orthologous cdc14hm allele was sufficient to elicit echinocandin hypersensitivity and perturb cell wall integrity signaling. It also caused striking abnormalities in septum structure and the same cell separation and hyphal differentiation phenotypes previously observed with cdc14 gene deletions. Since hyphal differentiation is important for C. albicans pathogenesis, we assessed the effect of reducing Cdc14 activity on virulence in Galleria mellonella and mouse models of invasive candidiasis. Partial reduction in Cdc14 activity via cdc14hm mutation severely impaired C. albicans virulence in both assays. Our results reveal that high Cdc14 activity promotes fungal cell wall integrity and, in C. albicans, is needed to orchestrate septation and hyphal differentiation, and for pathogenesis. Cdc14 may therefore be worth future exploration as an antifungal drug target.

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“…At least 100 cells were counted for each strain, experiment, and each time point shown in Figures 1F, 3, B and D, 4, B and E, and 5B. Specimens for electron microscopy were prepared as described previously (Meitinger and Pereira, 2016). …”

Section: Methodsmentioning

confidence: 99%

The septin-associated kinase Gin4 recruits Gps1 to the site of cell division

Meitinger

Pereira

2017

MBoC

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Visualization of Cytokinesis Events in Budding Yeast by Transmission Electron Microscopy

Cited by 4 publications

References 14 publications

Cdc14 phosphatase contributes to cell wall integrity and pathogenesis in Candida albicans

Cdc14 phosphatase contributes to cell wall integrity and pathogenesis in Candida albicans

Reduced Cdc14 phosphatase activity impairs septation, hyphal differentiation and pathogenesis and causes echinocandin hypersensitivity inCandida albicans

The septin-associated kinase Gin4 recruits Gps1 to the site of cell division

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