Protein phosphatase CaPpz1 is involved in cation homeostasis, cell wall integrity and virulence of Candida albicans

Ádám, Csaba; Erdei, Éva; Casado, Carlos; Kovács, László; González, Asier; Majoros, László; Petrényi, Katalin; Bagossi, Péter; Farkas, Illés J.; Molnár, Mónika; Pócsi, István; Ariño, Joaquı́n; Dombrádi, Viktor

doi:10.1099/mic.0.057075-0

Cited by 34 publications

(72 citation statements)

References 44 publications

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“…Complementation of RLM1 partially restored the virulence phenotype, indicating that gene dosage is important. This partial complementation has also been reported for several genes in C. albicans , including genes involved in the cell wall integrity [58], [60]–[62].…”

Section: Discussionsupporting

confidence: 73%

Participation of Candida albicans Transcription Factor RLM1 in Cell Wall Biogenesis and Virulence

et al. 2014

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Candida albicans cell wall is important for growth and interaction with the environment. RLM1 is one of the putative transcription factors involved in the cell wall integrity pathway, which plays an important role in the maintenance of the cell wall integrity. In this work we investigated the involvement of RLM1 in the cell wall biogenesis and in virulence. Newly constructed C. albicans Δ/Δrlm1 mutants showed typical cell wall weakening phenotypes, such as hypersensitivity to Congo Red, Calcofluor White, and caspofungin (phenotype reverted in the presence of sorbitol), confirming the involvement of RLM1 in the cell wall integrity. Additionally, the cell wall of C. albicans Δ/Δrlm1 showed a significant increase in chitin (213%) and reduction in mannans (60%), in comparison with the wild-type, results that are consistent with cell wall remodelling. Microarray analysis in the absence of any stress showed that deletion of RLM1 in C. albicans significantly down-regulated genes involved in carbohydrate catabolism such as DAK2, GLK4, NHT1 and TPS1, up-regulated genes involved in the utilization of alternative carbon sources, like AGP2, SOU1, SAP6, CIT1 or GAL4, and genes involved in cell adhesion like ECE1, ALS1, ALS3, HWP1 or RBT1. In agreement with the microarray results adhesion assays showed an increased amount of adhering cells and total biomass in the mutant strain, in comparison with the wild-type. C. albicans mutant Δ/Δrlm1 strain was also found to be less virulent than the wild-type and complemented strains in the murine model of disseminated candidiasis. Overall, we showed that in the absence of RLM1 the modifications in the cell wall composition alter yeast interaction with the environment, with consequences in adhesion ability and virulence. The gene expression findings suggest that this gene participates in the cell wall biogenesis, with the mutant rearranging its metabolic pathways to allow the use of alternative carbon sources.

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Section: Discussionsupporting

confidence: 73%

Participation of Candida albicans Transcription Factor RLM1 in Cell Wall Biogenesis and Virulence

et al. 2014

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“…Like other fungus-specific phosphatases, C. albicans PPZ1 (484 amino acids [aa], 54.4 kDa) has two domains: an N-terminal intrinsically disordered protein (IDP) domain (aa 1 to 170 [PPZ1 Nterm ]) and a C-terminal catalytic domain (aa 171 to 484 [PPZ1 cat ]) (14, 16). PPZ1 cat and human PP1α exhibit 66% sequence identity throughout their catalytic domains, and the 6 residues that coordinate the active site metals are 100% conserved (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…GQ357913 [14, 36]) and cloned into the E. coli expression vector pGEX6p-1 (Amersham Biosciences); both constructs were sequence verified (UD Genomed, Ltd.). Both constructs were then transformed into E. coli BL21(DE3) RIL cells (Agilent), and expression of N-terminal glutathione S -transferase (GST)-tagged proteins was induced with 0.6 mM IPTG (Sigma) at 18°C.…”

Section: Methodsmentioning

confidence: 99%

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Molecular Insights into the Fungus-Specific Serine/Threonine Protein Phosphatase Z1 in Candida albicans

Chen

Choy

Petrényi

et al. 2016

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The opportunistic pathogen Candida is one of the most common causes of nosocomial bloodstream infections. Because candidemia is associated with high mortality rates and because the incidences of multidrug-resistant Candida are increasing, efforts to identify novel targets for the development of potent antifungals are warranted. Here, we describe the structure and function of the first member of a family of protein phosphatases that is specific to fungi, protein phosphatase Z1 (PPZ1) from Candida albicans. We show that PPZ1 not only is active but also is as susceptible to inhibition by the cyclic peptide inhibitor microcystin-LR as its most similar human homolog, protein phosphatase 1α (PP1α [GLC7 in the yeast Saccharomyces cerevisiae]). Unexpectedly, we also discovered that, despite its 66% sequence identity to PP1α, the catalytic domain of PPZ1 contains novel structural elements that are not present in PP1α. We then used activity and pulldown assays to show that these structural differences block a large subset of PP1/GLC7 regulatory proteins from effectively binding PPZ1, demonstrating that PPZ1 does not compete with GLC7 for its regulatory proteins. Equally important, these unique structural elements provide new pockets suitable for the development of PPZ1-specific inhibitors. Together, these studies not only reveal why PPZ1 does not negatively impact GLC7 activity in vivo but also demonstrate that the family of fungus-specific phosphatases—especially PPZ1 from C. albicans—are highly suitable targets for the development of novel drugs that specifically target C. albicans without cross-reacting with human phosphatases.

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“…Previous studies in yeast and higher eukaryotes have shown that PP2A-type phosphatases, in particular, control many cellular processes, including actin cytoskeleton organization, mitosis, DNA replication, cell wall integrity, cellular proliferation, apoptosis, gene expression, and signal transduction (35,37,(57)(58)(59). In the major human fungal pathogen Candida albicans, protein phosphatases are known to regulate cell cycle progression, cation homeostasis, cell wall integrity, macrophage killing, antifungal drug resistance (e.g., as mediated by calcineurin), osmotolerance, and the response to oxidative stress and DNA-damaging agents (33,(60)(61)(62)(63)(64)(65)(66). However, considerably less is known about the ability of phosphatases to control morphology, one of the most important C. albicans virulence traits.…”

Section: Discussionmentioning

confidence: 99%

Ppg1, a PP2A-Type Protein Phosphatase, Controls Filament Extension and Virulence in Candida albicans

et al. 2014

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b Candida albicans, a major human fungal pathogen, is the primary cause of invasive candidiasis in a wide array of immunocompromised patients. C. albicans virulence requires the ability to undergo a reversible morphological transition from yeast to filaments in response to a variety of host environmental cues. These cues are sensed by the pathogen and activate multiple signal transduction pathways to induce filamentation. Reversible phosphorylation events are critical for regulation of many of these pathways. While a variety of protein kinases are known to function as components of C. albicans filamentous growth signal transduction pathways, considerably little is known about the role of phosphatases. Here we demonstrate that PPG1, encoding a putative type 2A-related protein phosphatase, is important for C. albicans filament extension, invasion, and virulence in a mouse model of systemic candidiasis. PPG1 is also important for downregulation of NRG1, a key transcriptional repressor of C. albicans filamentous growth, and is shown to affect the expression of several filament-specific target genes. An epistasis analysis suggests that PPG1 controls C. albicans filamentation via the cyclic AMP-protein kinase A (cAMP-PKA) signaling pathway. We demonstrate that Ppg1 possesses phosphatase activity and that a ppg1 catalytic mutant shows nearly equivalent filamentation, invasion, and virulence defects compared to those of a ppg1⌬/⌬ strain. Overall, our results suggest that phosphatases, such as Ppg1, play critical roles in controlling and fine-tuning C. albicans filament extension and virulence as well as signal transduction pathways, transcriptional regulators, and target genes associated with these processes.

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Protein phosphatase CaPpz1 is involved in cation homeostasis, cell wall integrity and virulence of Candida albicans

Cited by 34 publications

References 44 publications

Participation of Candida albicans Transcription Factor RLM1 in Cell Wall Biogenesis and Virulence

Participation of Candida albicans Transcription Factor RLM1 in Cell Wall Biogenesis and Virulence

Molecular Insights into the Fungus-Specific Serine/Threonine Protein Phosphatase Z1 in Candida albicans

Ppg1, a PP2A-Type Protein Phosphatase, Controls Filament Extension and Virulence in Candida albicans

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