Generating genomic platforms to study Candida albicans pathogenesis

Legrand, Mélanie; Bachellier‐Bassi, Sophie; Kk, Lee; Chaudhari, Yogesh; Tournu, Hélène; Arbogast, Laurence; Boyer, Hélène; Chauvel, Murielle; Cabral, Pedro M.; Maufrais, Corinne; Nesseir, Audrey; Maslanka, Irena; Permal, Emmanuelle; Rossignol, Tristan; Walker, Louise A.; Zeidler, Ute; Znaidi, Sadri; Schoeters, Floris; Majgier, Charlotte; Ra, Julien; Ma, Langlang; Tichit, Magalie; Bouchier, Christiane; Dijck, Patrick Van; Ca, Munro; d’Enfert, Christophe

doi:10.1093/nar/gky747

Cited by 8 publications

(30 citation statements)

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“…With the available ORFeome collection ( 32 ), a preliminary, random library of prey-harboring C. albicans strains was generated. Using the LR reaction (Gateway system), 1,646 open reading frames (ORFs) were transferred from the pDONR207-XX (XX denoting a certain ORF) plasmids to the prey plasmid pC2HP-GC ( 32 ). Resulting prey plasmids pC2HP-XX were then transformed into mating-compatible strain SC2H3a-pWOR1.…”

Section: Resultsmentioning

confidence: 99%

“…However, the system was limited to a low-scale setup ( 19 ). To overcome this limitation, the original C2H vectors were made Gateway compatible and inducible mating-competent C. albicans strains were created ( 32 ). The latter is important since the low transformation efficiency and unstable episomal plasmids make it difficult to perform a traditional pooled-style PPI screen.…”

Section: Resultsmentioning

confidence: 99%

“…The latter is important since the low transformation efficiency and unstable episomal plasmids make it difficult to perform a traditional pooled-style PPI screen. In the previously described study ( 32 ), the mating-compatible diploid strains harboring a bait or prey plasmid were mixed in liquid Spider medium without the addition of doxycycline (Dox) (see also below) ( 32 ). This approach is, however, less feasible for a high-throughput setup since mating in 96-well plates in liquid culture proved to be inefficient.…”

Section: Resultsmentioning

confidence: 99%

“…Prey- and bait-harboring strains were manually transferred with a 96-replicate pinner, respectively, from SC-Arg and SC-Leu to Spider-plus-Dox agar plates and incubated for 5 to 6 days at 23°C to induce mating and the generation of tetraploids. The addition of doxycycline (Dox) is necessary for the induction of opaque switching by the WOR1 regulator to stimulate mating by forming opaque cells ( 32 ). When doxycycline was omitted from the Spider medium, the mating efficiency dropped substantially even if Dox was added in the precultures of the prey and baits on SC-Arg or SC-Leu agar.…”

Section: Resultsmentioning

confidence: 99%

“…With the sequencing of the whole genome ( 16 , 29 – 31 ), the availability of a Gateway-adapted C. albicans ORFeome containing 83% of the currently annotated coding sequences, and the adaption of the C2H system to a high-throughput setup, it became possible to screen PPIs on a larger scale ( 32 ). Here, we show the potential of the C2H system to detect PPIs by presenting the first large-scale application of this C2H system.…”

Section: Introductionmentioning

confidence: 99%

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A High-Throughput Candida albicans Two-Hybrid System

Schoeters

Munro

d’Enfert

et al. 2018

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Candida albicans is a major fungal pathogen, and due to the rise of fungal infections and emerging resistance to the limited antifungals available, it is important to develop novel and more specific antifungals. Protein-protein interactions (PPIs) can be applied as very specific drug targets. However, because of the aberrant codon usage of C. albicans, the traditional yeast two-hybrid system in Saccharomyces cerevisiae is difficult to use, and only a limited number of PPIs have been described in C. albicans. To overcome this, a C. albicans two-hybrid (C2H) system was developed in 2010. The current work describes, for the first time, the application of the C2H system in a high-throughput setup. We hereby show the usefulness of the C2H system to investigate and detect PPIs in C. albicans, making it possible to further elucidate protein networks in C. albicans, which has the potential to lead to the development of novel antifungals which specifically disrupt PPIs important for virulence.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

A High-Throughput Candida albicans Two-Hybrid System

Schoeters

Munro

d’Enfert

et al. 2018

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Candida albicans: An Emerging Yeast Model to Study Eukaryotic Genome Plasticity

et al. 2019

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HighlightsC. albicans is a major human pathogen, with features that distinguish it from other yeast species.Rather than meiosis, ploidy reduction upon mating occurs by a parasexual process of concerted chromosome loss.Centromeres have unique organisation and are epigenetically regulated. Neither centromeres nor the DNA replication origins share any common defining DNA sequence.Loss-of-heterozygosity events are frequent and widespread in the genome of C. albicans.The DNA damage response is coupled to morphogenetic shifts.C. albicans emerges as a new unicellular model to study eukaryotic genome biology and dynamics.

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A conserved regulator controls asexual sporulation in the fungal pathogen Candida albicans

et al. 2020

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Transcription factor Rme1 is conserved among ascomycetes and regulates meiosis and pseudohyphal growth in Saccharomyces cerevisiae. The genome of the meiosis-defective pathogen Candida albicans encodes an Rme1 homolog that is part of a transcriptional circuitry controlling hyphal growth. Here, we use chromatin immunoprecipitation and genome-wide expression analyses to study a possible role of Rme1 in C. albicans morphogenesis. We find that Rme1 binds upstream and activates the expression of genes that are upregulated during chlamydosporulation, an asexual process leading to formation of large, spherical, thick-walled cells during nutrient starvation. RME1 deletion abolishes chlamydosporulation in three Candida species, whereas its overexpression bypasses the requirement for chlamydosporulation cues and regulators. RME1 expression levels correlate with chlamydosporulation efficiency across clinical isolates. Interestingly, RME1 displays a biphasic pattern of expression, with a first phase independent of Rme1 function and dependent on chlamydospore-inducing cues, and a second phase dependent on Rme1 function and independent of chlamydospore-inducing cues. Our results indicate that Rme1 plays a central role in chlamydospore development in Candida species.

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Generating genomic platforms to study Candida albicans pathogenesis

Cited by 8 publications

References 0 publications

A High-Throughput Candida albicans Two-Hybrid System

A High-Throughput Candida albicans Two-Hybrid System

Candida albicans: An Emerging Yeast Model to Study Eukaryotic Genome Plasticity

A conserved regulator controls asexual sporulation in the fungal pathogen Candida albicans

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