An amino acid liquid synthetic medium for the development of mycellal and yeast forms of<i>Candida albicans</i>

Lee, K.L.; Buckley, Helen R.; Campbell, Charlotte C.

doi:10.1080/00362177585190271

Cited by 798 publications

(514 citation statements)

References 11 publications

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“…Ura 2 auxotrophs were selected on SD with 1 mg 5-fluoro-orotic acid ml 21 and 25 mg uridine ml 21 . The medium for pH induction of hyphae was according to Lee et al (1975); Spider and salt base N-acetylglucosamine (GlcNAc)-containing media were according to Liu et al (1994) and Delbrück & Ernst (1993), respectively. All media were routinely prepared with uridine in order to use them for Ura 2 strains as well.…”

Section: Methodsmentioning

confidence: 99%

The cAMP phosphodiesterase encoded by CaPDE2 is required for hyphal development in Candida albicans

Jung

Stateva

2003

Microbiology

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The cAMP-dependent pathway, which regulates yeast-to-hypha morphogenesis in Candida albicans, is controlled by changes in cAMP levels determined by the processes of synthesis and hydrolysis. Both low-and high-affinity cAMP phosphodiesterases are encoded in the C. albicans genome. CaPDE2, encoding the high-affinity cAMP phosphodiesterase, has been cloned and shown to be toxic in Saccharomyces cerevisiae upon overexpression under pGAL1, but functional under the moderate pMET3. Deletion of CaPDE2 causes elevated cAMP levels and responsiveness to exogenous cAMP, higher sensitivity to heat shock, severe growth defects at 42 6C and highly reduced levels of EFG1 transcription. In vitro in hypha-inducing liquid medium CaPDE2, deletion prohibits normal hyphal, but not pseudohyphal growth. On solid medium capde2 mutants form aberrant hyphae, with fewer branches and almost no lateral buds, which are deficient in hypha-to-yeast reversion. The phenotypic defects of capde2 mutants show that the cAMP-dependent pathway plays specific roles in hyphal and pseudohyphal development, its regulatory role however, being greater in liquid than on solid medium in vitro. The increased expression of CaPDE2 after serum addition correlates well with a drop in cAMP levels following the initial rise in response to the hyphal inducer. These results suggest that Capde2p mediates a desensitization mechanism by lowering basal cAMP levels in response to environmental stimuli in C. albicans.

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Section: Methodsmentioning

confidence: 99%

The cAMP phosphodiesterase encoded by CaPDE2 is required for hyphal development in Candida albicans

Jung

Stateva

2003

Microbiology

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“…Control cultures were grown in fresh Soll's Medium at 25 "C and pH 4.5, 25 OC and pH 6-5, and 37 "C and pH 4.5. Soll's Medium is the defined Lee et al (1975) modified according to Buffo et al (1984). Cell numbers and morphology were monitored by light microscopy.…”

Section: Methodsmentioning

confidence: 99%

Regulation of the gene encoding translation elongation factor 3 during growth and morphogenesis in Candida albicans

et al. 1994

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The level of the T€F3 mRNA, which encodes the fungal-specific translation elongation factor 3 (EF-3), was measured during the yeast-to-hyphal transition in Candida albicans. In contrast to a previous report, T€F3 mRNA levels were shown to change during dilution into fresh medium, increasing only transiently when dimorphism was induced by either (i) an increase in growth temperature (from 25 "C to 37 "C) combined with the addition of 10% (vh) bovine calf serum to the medium, or (ii) an increase in growth temperature (from 25 "C to 37 "C) combined with an increase in the pH of the medium (from pH 4 5 to 6.5). T€F3 mRNA levels also increased in control cultures under conditions where germ tubes were not formed, but they remained elevated in contrast t o cultures undergoing morphological changes. E F 3 mRNA levels were not significantly affected by heat-shock, but were tightly regulated during batch growth of the yeast form, reaching maximal levels in exponential phase. Therefore, the changes in T€F3 expression that accompany the dimorphic transition in C. albicans appear to reflect the underlying physiological changes that occur during morphogenesis and are not a response to morphogenesis per se. For this reason T€F3 mRNA measurement cannot be used as a loading control in Northern analyses of dimorphic gene regulation. Comparison of TEF3 mRNA levels with the abundance of the EF-3 polypeptide indicated that the synthesis of this essential translation factor might be subject to post-transcriptional regulation.

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“…Depending on experimental conditions, C. albicans strains were grown in YPD (1% yeast extract, 2% peptone, and 1% dextrose), YP (1% yeast extract, 2% peptone) supplemented with 10% Fetal Bovine Serum (FBS), SD (synthetic dextrose, 0.67% yeast nitrogen base (YNB; Difco) with 2% glucose) [69] supplemented if necessary with arginine, histidine or uridine (20 mg/l each and 2% agar for growth on solid medium), SC (synthetic complete) or Lee's medium supplemented or not with methionine [70]. Expression from the tetracycline-inducible promoter (P TET ) was achieved through addition of 3 mg/ml anhydrotetracycline (ATcFisher Bioblock Scientific) in YPD at 30uC [41].…”

Section: Strains and Growth Mediamentioning

confidence: 99%

A comprehensive functional portrait of two heat shock factor-type transcriptional regulators involved in Candida albicans morphogenesis and virulence

Znaidi

Nesseir

Chauvel

et al. 2013

Journal de Mycologie Médicale

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Sfl1p and Sfl2p are two homologous heat shock factor-type transcriptional regulators that antagonistically control morphogenesis in Candida albicans, while being required for full pathogenesis and virulence. To understand how Sfl1p and Sfl2p exert their function, we combined genome-wide location and expression analyses to reveal their transcriptional targets in vivo together with the associated changes of the C. albicans transcriptome. We show that Sfl1p and Sfl2p bind to the promoter of at least 113 common targets through divergent binding motifs and modulate directly the expression of key transcriptional regulators of C. albicans morphogenesis and/or virulence. Surprisingly, we found that Sfl2p additionally binds to the promoter of 75 specific targets, including a high proportion of hyphal-specific genes (HSGs; HWP1, HYR1, ECE1, others), revealing a direct link between Sfl2p and hyphal development. Data mining pointed to a regulatory network in which Sfl1p and Sfl2p act as both transcriptional activators and repressors. Sfl1p directly represses the expression of positive regulators of hyphal growth (BRG1, UME6, TEC1, SFL2), while upregulating both yeast form-associated genes (RME1, RHD1, YWP1) and repressors of morphogenesis (SSN6, NRG1). On the other hand, Sfl2p directly upregulates HSGs and activators of hyphal growth (UME6, TEC1), while downregulating yeast form-associated genes and repressors of morphogenesis (NRG1, RFG1, SFL1). Using genetic interaction analyses, we provide further evidences that Sfl1p and Sfl2p antagonistically control C. albicans morphogenesis through direct modulation of the expression of important regulators of hyphal growth. Bioinformatic analyses suggest that binding of Sfl1p and Sfl2p to their targets occurs with the co-binding of Efg1p and/or Ndt80p. We show, indeed, that Sfl1p and Sfl2p targets are bound by Efg1p and that both Sfl1p and Sfl2p associate in vivo with Efg1p. Taken together, our data suggest that Sfl1p and Sfl2p act as central ''switch on/off'' proteins to coordinate the regulation of C. albicans morphogenesis.

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An amino acid liquid synthetic medium for the development of mycellal and yeast forms ofCandida albicans

Cited by 798 publications

References 11 publications

The cAMP phosphodiesterase encoded by CaPDE2 is required for hyphal development in Candida albicans

The cAMP phosphodiesterase encoded by CaPDE2 is required for hyphal development in Candida albicans

Regulation of the gene encoding translation elongation factor 3 during growth and morphogenesis in Candida albicans

A comprehensive functional portrait of two heat shock factor-type transcriptional regulators involved in Candida albicans morphogenesis and virulence

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